MATSUMOTO Kazuya

    Department of Genetic Engineering Professor
Last Updated :2024/04/25

Researcher Information

Degree

  • (BLANK)(1989/03 Kyoto University)

Research funding number

  • 20298938

J-Global ID

Research Interests

  • バイオインフォマティクス   分子発生生物学   生殖生物学   Molecular Developmental Biology   Reproductive Biology   

Research Areas

  • Life sciences / Animal production science
  • Life sciences / Laboratory animal science
  • Life sciences / Animals: biochemistry, physiology, behavioral science

Education

  •        - 1989/03  Kyoto University  農学研究科  博士後期課程修了(畜産学専攻)
  •        - 1989  Kyoto University  Graduate School, Division of Agriculture
  •        - 1986/03  Kyoto University  農学研究科  博士前期課程修了(畜産学専攻)
  •        - 1983/03  Utsunomiya University  Faculty of Agriculture  畜産学科 卒業
  •        - 1983  Utsunomiya University  Faculty of Agriculture

Association Memberships

  • 日本受精着床学会   日本卵子学会   日本繁殖生物学会   日本畜産学会   

Published Papers

  • 精管灌流によるアカカンガルー精子回収および形態学的観察の試み
    安齋 政幸; 由良 晶子; 尾崎 美樹; 野田 義博; 藤野 奈央; 大原 なずな; 田中 万柚子; 網師本 沙恵; 松本 和也
    日本野生動物医学会誌 日本野生動物医学会 27 (Suppl.) 187 - 188 1342-6133 2023/03
  • 体外成熟培養に供試したヒト卵のトランスクリプトーム解析
    山本 真理; 武内 大輝; 福井 愛実; 前沢 忠志; 西岡 美喜子; 池田 智明; 松本 和也; 宮本 圭
    日本生殖医学会雑誌 (一社)日本生殖医学会 67 (3) 134 - 134 1881-0098 2022/07 [Refereed]
  • Shunya Ihashi; Mizuto Hamanaka; Masaya Kaji; Miki Mori; Yuma Imasato; Misaki Nakamura; Masayuki Anzai; Kazuya Matsumoto; Masahito Ikawa; Kei Miyamoto
    bioRxiv Cold Spring Harbor Laboratory 2022/04 
    SUMMARY Differentiated cell nuclei can be reprogrammed after nuclear transfer (NT) to oocytes and the produced NT embryos can give rise to cloned animals. However, development of NT embryos is often hampered by recurrent reprogramming failures, including the incomplete activation of developmental genes, yet specific genes responsible for the arrest of NT embryos are not well understood. Here, we searched for developmentally important genes among the reprogramming-resistant H3K9me3-repressed genes, and identified Alyref and Gabpb1 by siRNA screening. Gene knockout of Alyref and Gabpb1 by the CRISPR/Cas9 system resulted in early developmental arrest in mice. Single embryo RNA-seq revealed that Alyref is needed for the formation of inner cell mass. The supplement of Alyref and Gabpb1 by mRNA injection supported efficient preimplantation development of cloned embryos. Thus, our study shows that the H3K9me3-repressed genes contain developmentally required genes and the incomplete activation of such genes results in preimplantation arrest of cloned embryos.
  • Hiroki Takeuchi; Mari Yamamoto; Megumi Fukui; Akihiro Inoue; Tadashi Maezawa; Mikiko Nishioka; Eiji Kondo; Tomoaki Ikeda; Kazuya Matsumoto; Kei Miyamoto
    Reproductive medicine and biology 21 (1) e12464  2022 
    PURPOSE: In vitro maturation (IVM) of human oocytes offers an invaluable opportunity for infertility treatment. However, in vitro matured oocytes often show lower developmental abilities than their in vivo counterparts, and molecular mechanisms underlying successful maturation remain unclear. In this study, we investigated gene expression profiles of in vitro matured oocytes at the single-cell level to gain mechanistic insight into IVM of human oocytes. METHODS: Human oocytes were retrieved by follicular puncture and in vitro matured. In total, 19 oocytes from 11 patients were collected and subjected to single-cell RNA-seq analyses. RESULTS: Global gene expression profiles were similar among oocytes at the same maturation stage, while a small number of oocytes showed distinct transcriptomes from those at the corresponding maturation stage. Differential gene expression analysis identified hundreds of transcripts that dynamically altered their expression during IVM, and we revealed molecular pathways and upstream regulators that may govern oocyte maturation. Furthermore, oocytes that were delayed in their maturation showed distinct transcriptomes. Finally, we identified genes whose transcripts were enriched in each stage of oocyte maturation. CONCLUSIONS: Our work uncovers transcriptomic changes during human oocyte IVM and the differential gene expression profile of each oocyte.
  • Junko Tomikawa; Christopher A Penfold; Takuma Kamiya; Risa Hibino; Ayumi Kosaka; Masayuki Anzai; Kazuya Matsumoto; Kei Miyamoto
    iScience 24 (11) 103290 - 103290 2021/11 [Refereed]
     
    Nuclear transfer systems represent the efficient means to reprogram a cell and in theory provide a basis for investigating the development of endangered species. However, conventional nuclear transfer using oocytes of laboratory animals does not allow reprogramming of cross-species nuclei owing to defects in cell divisions and activation of embryonic genes. Here, we show that somatic nuclei transferred into mouse four-cell embryos arrested at the G2/M phase undergo reprogramming toward the embryonic state. Remarkably, genome-wide transcriptional reprogramming is induced within a day, and ZFP281 is important for this replication-free reprogramming. This system further enables transcriptional reprogramming of cells from Oryx dammah, now extinct in the wild. Thus, our findings indicate that arrested mouse embryos are competent to induce intra- and cross-species reprogramming. The direct induction of embryonic transcripts from diverse genomes paves a unique approach for identifying mechanisms of transcriptional reprogramming and genome activation from a diverse range of species.
  • Kohtaro Morita; Yuki Hatanaka; Shunya Ihashi; Masahide Asano; Kei Miyamoto; Kazuya Matsumoto
    Scientific reports 11 (1) 10146 - 10146 2021/05 [Refereed]
     
    Paternal genome reprogramming, such as protamine-histone exchange and global DNA demethylation, is crucial for the development of fertilised embryos. Previously, our study showed that one of histone arginine methylation, asymmetrically dimethylated histone H3R17 (H3R17me2a), is necessary for epigenetic reprogramming in the mouse paternal genome. However, roles of histone arginine methylation in reprogramming after fertilisation are still poorly understood. Here, we report that H3R2me2s promotes global transcription at the 1-cell stage, referred to as minor zygotic genome activation (ZGA). The inhibition of H3R2me2s by expressing a histone H3.3 mutant H3.3R2A prevented embryonic development from the 2-cell to 4-cell stages and significantly reduced global RNA synthesis and RNA polymerase II (Pol II) activity. Consistent with this result, the expression levels of MuERV-L as minor ZGA transcripts were decreased by forced expression of H3.3R2A. Furthermore, treatment with an inhibitor and co-injection of siRNA to PRMT5 and PRMT7 also resulted in the attenuation of transcriptional activities with reduction of H3R2me2s in the pronuclei of zygotes. Interestingly, impairment of H3K4 methylation by expression of H3.3K4M resulted in a decrease of H3R2me2s in male pronuclei. Our findings suggest that H3R2me2s together with H3K4 methylation is involved in global transcription during minor ZGA in mice.
  • Tomomi Okuno; Wayne Yang Li; Yu Hatano; Atsushi Takasu; Yuko Sakamoto; Mari Yamamoto; Zenki Ikeda; Taiki Shindo; Matthias Plessner; Kohtaro Morita; Kazuya Matsumoto; Kazuo Yamagata; Robert Grosse; Kei Miyamoto
    Cell reports 31 (13) 107824 - 107824 2020/06 [Refereed]
     
    After fertilization, sperm and oocyte nuclei are rapidly remodeled to form swollen pronuclei (PN) in mammalian zygotes, and the proper formation and function of PN are key to producing totipotent zygotes. However, how mature PN are formed has been unclear. We find that filamentous actin (F-actin) assembles in the PN of mouse zygotes and is required for fully functional PN. The perturbation of nuclear actin dynamics in zygotes results in the misregulation of genes related to genome integrity and abnormal development of mouse embryos. We show that nuclear F-actin ensures DNA damage repair, thus preventing the activation of a zygotic checkpoint. Furthermore, optogenetic control of cofilin nuclear localization reveals the dynamically regulated F-actin nucleoskeleton in zygotes, and its timely disassembly is needed for developmental progression. Nuclear F-actin is a hallmark of totipotent zygotic PN, and the temporal regulation of its polymerized state is necessary for normal embryonic development.
  • Rika Azuma; Yuki Hatanaka; Seung-Wook Shin; Hitoshi Murai; Minoru Miyashita; Masayuki Anzai; Kazuya Matsumoto
    The Journal of reproduction and development 66 (3) 255 - 263 2020/06 [Refereed]
     
    The large Japanese field mouse (Apodemus speciosus) is endemic to Japan and may be used as an animal model for studies related to environmental pollution, medical science, and basic biology. However, the large Japanese field mouse has low reproductive ability due to the small number of oocytes ovulated per female. To produce experimental models, we investigated the in vitro developmental potential of interspecies somatic cell nuclear transfer (iSCNT) embryos produced by fusing tail tip cells from the large Japanese field mouse with enucleated oocytes from laboratory mice (Mus musculus domesticus). Only a small number of iSCNT embryos developed to the 4-cell (0-4%) and blastocysts (0-1%) stages under sequential treatment using trichostatin A (TSA) and vitamin C (VC) supplemented with deionized bovine serum albumin (d-BSA). This sequential treatment led to the reduction in H3K9 trimethylation and did not affect H3K4 trimethylation in at least the 2-cell stage of the iSCNT embryos. Moreover, iSCNT embryos that received tail tip cells with exposure treatment to ooplasm from cell fusion to oocyte activation or VC treatment prior to cell fusion did not exhibit significant in vitro development improvement compared to that of each control group. This suggests that large Japanese field mice/laboratory mice iSCNT embryos that received sequential treatment using TSA and VC with d-BSA may have slightly better developmental potential beyond the 4-cell stage. Our results provide insights into the reprogramming barriers impeding the wider implementation of iSCNT technology.
  • 松本和也
    日本畜産学会報 91 (3) 1346-907X 2020 [Refereed][Invited]
  • Higuchi C; Yamamoto M; Shin SW; Miyamoto K; Matsumoto K
    Biology open The Company of Biologists 8 (10) bio048652 - bio048652 2019/10 [Refereed]
  • 2万8千年前のケナガマンモス組織から得られたタンパク質の翻訳後修飾の解析
    西端 智也; 永井 宏平; 山縣 一夫; 宮本 裕史; 安齋 政幸; 加藤 博己; 宮本 圭; 東 里香; Kolodeznikov Igor I.; Protopopov Albert V.; Plotnikov Valerii V.; 細井 美彦; 三谷 匡; 松本 和也; 入谷 明
    電気泳動 日本電気泳動学会 63 (Suppl.) 195 - 195 2189-2628 2019/07
  • 2万8千年前のケナガマンモスの筋肉組織と骨髄組織のプロテオーム解析
    永井 宏平; 宮本 裕史; 安齋 政幸; 東 里香; 西端 智也; 山縣 一夫; 加藤 博己; 宮本 圭; Kolodeznikov Igor I.; Protopopov Albert V.; Plotnikov Valerii V.; 細井 美彦; 三谷 匡; 松本 和也; 入谷 明
    電気泳動 日本電気泳動学会 63 (Suppl.) 196 - 196 2189-2628 2019/07
  • Yamagata K; Nagai K; Miyamoto H; Anzai M; Kato H; Miyamoto K; Kurosaka S; Azuma R; Kolodeznikov II; Protopopov AV; Plotnikov VV; Kobayashi H; Kawahara-Miki R; Kono T; Uchida M; Shibata Y; Handa T; Kimura H; Hosoi Y; Mitani T; Matsumoto K; Iritani A
    Scientific reports 9 (1) 4050 - 4050 2019/03 [Refereed]
     
    The 28,000-year-old remains of a woolly mammoth, named 'Yuka', were found in Siberian permafrost. Here we recovered the less-damaged nucleus-like structures from the remains and visualised their dynamics in living mouse oocytes after nuclear transfer. Proteomic analyses demonstrated the presence of nuclear components in the remains. Nucleus-like structures found in the tissue homogenate were histone- and lamin-positive by immunostaining. In the reconstructed oocytes, the mammoth nuclei showed the spindle assembly, histone incorporation and partial nuclear formation; however, the full activation of nuclei for cleavage was not confirmed. DNA damage levels, which varied among the nuclei, were comparable to those of frozen-thawed mouse sperm and were reduced in some reconstructed oocytes. Our work provides a platform to evaluate the biological activities of nuclei in extinct animal species.
  • Nishihara T; Matsumoto K; Hosoi Y; Morimoto Y
    Reproductive medicine and biology 17 (4) 481 - 486 1445-5781 2018/10 [Refereed]
  • 受精卵およびクローン胚におけるエピジェネティックリプログラミング
    奥野智美; 松本和也; 宮本 圭
    日本胚移植学雑誌 40 (3) 117 - 122 2018/09 [Refereed][Invited]
  • 黒毛和種去勢牛の脂肪交雑を生体評価するバイオマーカー候補タンパク質の血清プロテオーム解析による探索
    池上春香; 松橋珠子; 永井宏平; 宮本圭; 大林賢伍; 坂口慎一; 松本和也
    関西畜産学会報 (175) 1 - 9 2018/03 [Refereed]
  • Higuchi C; Shimizu N; Shin SW; Morita K; Nagai K; Anzai M; Kato H; Mitani T; Yamagata K; Hosoi Y; Miyamoto K; Matsumoto K
    The Journal of reproduction and development Japanese Society of Animal Reproduction 64 (1) 65 - 74 0916-8818 2018/02 [Refereed]
  • Kohtaro Morita; Mikiko Tokoro; Yuki Hatanaka; Chika Higuchi; Haruka Ikegami; Kouhei Nagai; Masayuki Anzai; Hiromi Kato; Tasuku Mitani; Yoshitomo Taguchi; Kazuo Yamagata; Yoshihiko Hosoi; Kei Miyamoto; Kazuya Matsumoto
    Journal of Reproduction and Development The Japanese Society of Animal Reproduction (JSAR) 64 (2) 161 - 171 1348-4400 2018 [Refereed]
     
    Antioxidant mechanisms to adequately moderate levels of endogenous reactive oxygen species (ROS) are important for oocytes and embryos to obtain and maintain developmental competence, respectively. Immediately after fertilization, ROS levels in zygotes are elevated but the antioxidant mechanisms during the maternal-to-zygotic transition (MZT) are not well understood. First, we identified peroxiredoxin 1 (PRDX1) and PRDX2 by proteomics analysis as two of the most abundant endogenous antioxidant enzymes eliminating hydrogen peroxide (H2O2). We here report the cellular localization of hyperoxidized PRDX and its involvement in the antioxidant mechanisms of freshly fertilized oocytes. Treatment of zygotes at the pronuclear stage with H2O2 enhanced pronuclear localization of hyperoxidized PRDX in zygotes and concurrently impaired the generation of 5-hydroxymethylcytosine (5hmC) on the male genome, which is an epigenetic reprogramming event that occurs at the pronuclear stage. Thus, our results suggest that endogenous PRDX is involved in antioxidant mechanisms and epigenetic reprogramming during MZT.
  • 初期胚特異的レトロトランスポゾンMERVLの活性化に伴う核内変化
    奥野 智美; 山口 壮輝; 濱田 歩花; 竹林 真理愛; 守田 昂太郎; 樋口 智香; 神谷 拓磨; 安齋 政幸; 山縣 一夫; 細井 美彦; Jerome Jullien; 松本 和也; 宮本 圭
    生命科学系学会合同年次大会 生命科学系学会合同年次大会運営事務局 2017年度 [2P - 0708] 2017/12
  • Yuki Hatanaka; Takeshi Tsusaka; Natsumi Shimizu; Kohtaro Morita; Takehiro Suzuki; Shinichi Machida; Manabu Satoh; Arata Honda; Michiko Hirose; Satoshi Kamimura; Narumi Ogonuki; Toshinobu Nakamura; Kimiko Inoue; Yoshihiko Hosoi; Naoshi Dohmae; Toru Nakano; Hitoshi Kurumizaka; Kazuya Matsumoto; Yoichi Shinkai; Atsuo Ogura
    CELL REPORTS CELL PRESS 20 (12) 2756 - 2765 2211-1247 2017/09 [Refereed]
     
    At fertilization, the paternal genome undergoes extensive reprogramming through protamine-histone exchange and active DNA demethylation, but only a few maternal factors have been defined in these processes. We identified maternal Mettl23 as a protein arginine methyltransferase (PRMT), which most likely catalyzes the asymmetric dimethylation of histone H3R17 (H3R17me2a), as indicated by in vitro assays and treatment with TBBD, an H3R17 PRMT inhibitor. Maternal histone H3.3, which is essential for paternal nucleosomal assembly, is unable to be incorporated into the male pronucleus when it lacks R17me2a. Mettl23 interacts with Tet3, a 5mC-oxidizing enzyme responsible for active DNA demethylation, by binding to another maternal factor, GSE (gonad-specific expression). Depletion of Mettl23 from oocytes resulted in impaired accumulation of GSE, Tet3, and 5hmC in the male pronucleus, suggesting that Mettl23 may recruit GSE-Tet3 to chromatin. Our findings establish H3R17me2a and its catalyzing enzyme Mettl23 as key regulators of paternal genome reprogramming.
  • Kei Miyamoto; Yosuke Tajima; Koki Yoshida; Mami Oikawa; Rika Azuma; George E. Allen; Tomomi Tsujikawa; Tomomasa Tsukaguchi; Charles R. Bradshaw; Jerome Jullien; Kazuo Yamagata; Kazuya Matsumoto; Masayuki Anzai; Hiroshi Imai; John B. Gurdon; Masayasu Yamada
    BIOLOGY OPEN COMPANY OF BIOLOGISTS LTD 6 (4) 415 - 424 2046-6390 2017/04 [Refereed]
     
    Animal cloning has been achieved in many species by transplanting differentiated cell nuclei to unfertilized oocytes. However, the low efficiencies of cloning have remained an unresolved issue. Here we find that the combination of two small molecules, trichostatin A (TSA) and vitamin C (VC), under culture condition with bovine serum albumin deionized by ion-exchange resins, dramatically improves the cloning efficiency in mice and 15% of cloned embryos develop to term by means of somatic cell nuclear transfer (SCNT). The improvement was not observed by adding the non-treated, rather than deionized, bovine serum. RNA-seq analyses of SCNT embryos at the two-cell stage revealed that the treatment with TSA and VC resulted in the upregulated expression of previously identified reprogramming-resistant genes. Moreover, the expression of early-embryo-specific retroelements was upregulated by the TSA and VC treatment. The enhanced gene expression was relevant to the VC-mediated reduction of histone H3 lysine 9 methylation in SCNT embryos. Our study thus shows a simply applicable method to greatly improve mouse cloning efficiency, and furthers our understanding of how somatic nuclei acquire totipotency.
  • Tomoko Amano; Masayuki Anzai; Kazuya Matsumoto
    THERIOGENOLOGY ELSEVIER SCIENCE INC 86 (7) 1670 - 1684 0093-691X 2016/10 [Refereed]
     
    Here, we showed that the Clock gene was important for reproductive performance in mice. We compared outcomes from the four possible mating combinations between wild-type mice (WT) and mice homozygous for the Clock delta-19 mutation (CL). We found that the only significant differences were between the WT male x WT female and CL male x CL female mating groups; these groups differed with regard to elongation of the pregnancy period (19.3 vs. 20.5 days, respectively, P < 0.05) and the number of newborn pups (13.4 +/- 0.8 vs. 8.6 +/- 1.5, respectively, P < 0.05). Because CL dams impregnated by male CLs exhibited normal continuous increases in body weight during the entire gestation period and did not show any signs of spontaneous abortion from mid to late gestation, we reasoned that some embryos were lost before or at the time of implantation. Immediately before implantation (88 hours after fertilization), neither the number of embryos collected from uteri nor the percentage of the embryos that reached the blastocyst stage differed significantly among mating groups. In contrast, immediately after implantation (160 hours after fertilization), the average number of implantation sites was significantly lower for the CL male x CL female mating group than that for the WT male' x WT female mating group (7.0 vs.13.0, P < 0.05); this decrease was accompanied by a significant lowering of the positions of implantation sites in uteri, and this lowering of the implantation sites was more severe when mothers and embryos bore more CL alleles (WT male x WT female > CL male x WT female > WT male x CL female > CL male x CL female), suggesting that the Clock mutation reduced the reproduction performance of the parents by affecting the implantation capacity via such as embryos' ability to implant. (C) 2016 The Author(s). Published by Elsevier Inc.
  • Hironobu Sugimoto; Yuta Kida; Noriyoshi Oh; Kensaku Kitada; Kazuya Matsumoto; Kazuhiro Saeki; Takeshi Taniguchi; Yoshihiko Hosoi
    ZYGOTE CAMBRIDGE UNIV PRESS 23 (4) 494 - 500 0967-1994 2015/08 [Refereed]
     
    We examined growing oocytes collected from follicles remaining in superovulated rabbit ovaries, that were grown (in vitro growth, IVG) and matured (in vitro maturation, IVM) in vitro. We produced somatic cell nuclear transfer (SCNT) embryos using the mature oocytes and examined whether these embryos have the ability to develop to the blastocyst stage. In addition, we examined the effects of trichostatin A (TSA), a histone deacetylase inhibitor (HDACi), on the developmental competence of SCNT embryos derived from IVG-IVM oocytes. After growth for 7 days and maturation for 14-16 h in vitro, the growing oocytes reached the metaphase II stage (51.4%). After SCNT, these reconstructed embryos reached the blastocyst stage (20%). Furthermore, the rate of development to the blastocyst stage and the number of cells in the blastocysts in SCNT embryos derived from IVG-IVM oocytes were significantly higher for TSA-treated embryos compared with TSA-untreated embryos (40.6 versus 21.4% and 353.1 +/- 59.1 versus 202.5 +/- 54.6, P < 0.05). These results indicate that rabbit SCNT embryos using IVG-IVM oocytes have the developmental competence to reach the blastocyst stage.
  • Ikegami H; Nagai K; Matsuhashi T; Kobahashi N; Takemoto A; Yoshihiro T; Inoue E; Higuchi C; Morita K; Uchibori S; Amano T; Taguchi Y; Kato H; Iritani A; Matsumoto K
    Nihon Chikusan Gakkaiho Zootechnical Science Society of Japan 86 (2) 141 - 152 1880-8255 2015/06 [Refereed]
     
    Early assessment of carcass traits and meat quality characteristics during fattening is expected to improve the efficiency of beef cattle production. Yet, there have been few reports on developing an assessment index for feeding condition of Japanese Black cattle. In this study, to identify protein biomarkers for predicting carcass traits and meat quality characteristics of Japanese Black cattle by proteomics, we first compared protein expression profiles of perirenal white adipose tissue between low and high merit steers in five carcass traits (carcass weight, rib eye area, rib thickness, subcutaneous fat thickness, and beef marbling standard [BMS] number). We then investigated the involvement of the lower- or higher-expressed proteins in metabolic pathways and evaluated the expression of vimentin proteins by western blot analysis in individual steers. We identified 45 proteins which were significantly expressed in low or high merit groups of five carcass traits. Coordinate involvement of some identified proteins in glycolysis/gluconeogenesis and citrate cycle pathways was found in high merit groups of carcass weight, subcutaneous fat thickness and BMS number. The expression of vimentin protein, which was significantly expressed in the high BMS number group, was confirmed in individual steers. Taken together, these results suggest the possibility of these identified proteins as protein biomarker candidates for predicting carcass traits and meat quality characteristics during fattening of Japanese Black cattle.
  • 池上春香; 永井宏平; 松橋珠子; 小林直彦; 武本淳史; 吉廣卓哉; 井上悦子; 樋口智香; 守田昂太郎; 内堀翔; 天野朋子; 田口善智; 加藤博己; 入谷明; 松本和也
    日本畜産学会報 Zootechnical Science Society of Japan 86 (2) 141 - 152 1346-907X 2015/05 [Refereed]
     
    Early assessment of carcass traits and meat quality characteristics during fattening is expected to improve the efficiency of beef cattle production. Yet, there have been few reports on developing an assessment index for feeding condition of Japanese Black cattle. In this study, to identify protein biomarkers for predicting carcass traits and meat quality characteristics of Japanese Black cattle by proteomics, we first compared protein expression profiles of perirenal white adipose tissue between low and high merit steers in five carcass traits (carcass weight, rib eye area, rib thickness, subcutaneous fat thickness, and beef marbling standard [BMS] number). We then investigated the involvement of the lower- or higher-expressed proteins in metabolic pathways and evaluated the expression of vimentin proteins by western blot analysis in individual steers. We identified 45 proteins which were significantly expressed in low or high merit groups of five carcass traits. Coordinate involvement of some identified proteins in glycolysis/gluconeogenesis and citrate cycle pathways was found in high merit groups of carcass weight, subcutaneous fat thickness and BMS number. The expression of vimentin protein, which was significantly expressed in the high BMS number group, was confirmed in individual steers. Taken together, these results suggest the possibility of these identified proteins as protein biomarker candidates for predicting carcass traits and meat quality characteristics during fattening of Japanese Black cattle.
  • 塚口 智将; 守田 昴太郎; 宮本 圭; 松本和也
    Memoirs of Institute of Advanced Technology, Kinki University 近畿大学先端技術総合研究所 6号 (20) 1 - 8 1346-8693 2015/03 [Refereed]
     
    [要旨] 哺乳類の初期胚では, 分化全能性を獲得するために, 受精直後に終末分化した精子と卵子のエピゲノムの情報がリセットされる. この現象では, 遺伝子の転写制御に関与するDNA やヒストンのメチル化及びアセチル化修飾がダイナミックに変化することが知られている. 受精後, 遺伝子の転写抑制に関わるDNAのメチル化修飾は, 酵素及びDNA 複製によって取り除かれ, その後再び, DNA メチル基転移酵素(DNAmethyltransferases, DNMTs)が働くことにより, 特定の遺伝子の転写制御が行われる. ヒストンにおいては, histone methyltransferase(HMT)によるメチル化, histone acetyltransferase(HAT)によるアセチル化などの修飾がクロマチン構造の変化を誘起させ, 転写の活性あるいは抑制に関わっている. 以上のように遺伝子の発現は, DNA やヒストン修飾の相互の働きかけによって制御されていると考えられる. したがって, 初期胚で起こるDNA 及びヒストン修飾の機構やそれらの修飾の持つ役割を知ることで, エピジェネティックリプログラミングの詳細な分子機構を深く理解することができる. さらに, それらの知見をiPS細胞作製効率の向上や分化誘導の改善に応用させることで, 再生医療や創薬分野への貢献が期待される. そこで本稿では, 哺乳類の初期胚において発生に必要な遺伝子の転写制御に関与しているDNA 及びヒストンのメチル化修飾について概説する. [Abstract] In mammalian early embryo, epigenome information of sperm and oocytes is reset to acquire the developmental totipotency. This phenomenon is accompanied by the dynamic changes in DNA and histone modifications. DNA methylation involved in transcriptional repression is removed by the enzymes or DNA replication after fertilization. The genome is remethylated by DNA methyltransferases(DNMTs) and the specific set of genes is transcriptionally repressed. Histone modifications such as methylation by histone methyltransferases(HMTs) and acetylation by histone acetyltransferases(HATs) also induce changes in chromatin structure and are involved in transcriptional regulation. Therefore, gene expression regulation seems to be achieved by the combination of DNA and histone modifications. Consequently, understanding mechanisms and roles of DNA and histone modifications can be a clue to understand detailed mechanism of epigenetic reprogramming. Moreover, the basic knowledge about epigenetic reprogramming to improve iPS cell conversion and induction of differentiation would contribute to the fields of regenerative medicine and innovative drug development. Here, we will review DNA and histone methylation related totranscriptional regulation of genes necessary for development in mammalian early embryo.近畿大学先端技術総合研究所紀要編集委員会
  • Natsumi Shimizu; Kimihiro Ueno; Ena Kurita; Seung-Wook Shin; Takuji Nishihara; Tomoko Amano; Masayuki Anzai; Satoshi Kishigami; Hiromi Kato; Tasuku Mitani; Yoshihiko Hosoi; Kazuya Matsumoto
    JOURNAL OF REPRODUCTION AND DEVELOPMENT SOCIETY REPRODUCTION & DEVELOPMENT-SRD 60 (3) 179 - 186 0916-8818 2014/06 [Refereed]
     
    In the mammalian testis, the ubiquitin-proteasome system plays important roles in the process that promotes the formation of mature sperm. We recently identified zygote-specific proteasome assembly chaperone (ZPAC), which is specifically expressed in the mouse gonads and zygote. ZPAC mediates a unique proteasome assembly pathway in the zygote, but the expression profile and function of ZPAC in the testis is not fully understood. In this study, we investigated the possible role of ZPAC during mouse spermatogenesis. First, we analyzed the expression of ZPAC and 20S proteasome subunit alpha 4/PSMA7 in the adult mouse testis. ZPAC and alpha 4 were expressed in spermatogonia, spermatocytes, and round spermatids. In elongating spermatids, ZPAC was expressed until step 10, whereas expression of alpha 4 persisted until step 12. We then examined the expression profile of ZPAC and alpha 4 in a mouse model of experimental unilateral cryptorchidism. Consistent with appearance of morphologically impaired germ cells following cryptorchidism, the ZPAC protein level was significantly decreased at 4 days post induction of experimental cryptorchidism (D4) compared with the intact testis, although the amount of alpha 4 protein persisted at least until D10. Moreover, intense ZPAC staining was co-localized with staining of annexin V, an early indicator of apoptosis in mammalian cells, in germ cells of cryptorchid testis, but ZPAC was also expressed in germ cells showing no detectable expression of annexin V. These results suggest that ZPAC plays a role during spermatogenesis and raises the possibility that 20S proteasome mediated by ZPAC may be involved in the regulation of germ cell survival during spermatogenesis.
  • Takuji Nishihara; Shu Hashimoto; Keijiro Ito; Yoshiharu Nakaoka; Kazuya Matsumoto; Yoshihiko Hosoi; Yoshiharu Morimoto
    GYNECOLOGICAL ENDOCRINOLOGY INFORMA HEALTHCARE 30 (5) 359 - 362 0951-3590 2014/05 [Refereed]
     
    The aim of this study was to evaluate the efficacy of oral melatonin supplementation on oocyte and embryo quality in patients in an assisted reproductive technologies program. All patients were treated for at least 2 weeks with melatonin (3 mg/day). To evaluate the cumulative effect of melatonin supplementation, we compared cycle outcomes between the first (no supplementation) and second cycles (melatonin supplementation) of patients who completed two treatment cycles. There were no significant differences in maturation rates (p = 0.50), blastocyst rates (p = 0.75), and the rate of good quality blastocysts (p = 0.59) between the first and second cycles. The fertilization rate of ICSI was higher in the second cycle than that in the first cycle (69.3 versus 77.5%). Being limited to patients with a low fertilization rate in the first cycle (<60%), the fertilization rate dramatically increased after melatonin treatment (35.1 versus 68.2%). The rate of good quality embryos also increased (48.0 versus 65.6%). An important finding in our study was that oral melatonin supplementation can have a beneficial effect on the improvement of fertilization and embryo quality and this may have occurred due to a reduction in oxidative damage.
  • Nagai Kouhei; Ikegami Haruka; Matsuhashi Tamako; Takemoto Atsushi; Minakata Yusuke; Higuchi Chika; Morita Koutaro; Kobayashi Naohiko; Matsumoto Kazuya
    Abstracts for Annual Meeting of Japanese Proteomics Society Japanese Proteomics Society (Japan Human Proteome Organisation) 2014 98 - 98 2014
  • Takayuki Yamochi; Yuta Kida; Noriyoshi Oh; Sei Ohta; Tomoko Amano; Masayuki Anzai; Hiromi Kato; Satoshi Kishigami; Tasuku Mitani; Kazuya Matsumoto; Kazuhiro Saeki; Makoto Takenoshita; Akira Iritani; Yoshihiko Hosoi
    ZYGOTE CAMBRIDGE UNIV PRESS 21 (4) 358 - 366 0967-1994 2013/11 [Refereed]
     
    Interspecies somatic cell nuclear transfer (ISCNT) has been proposed as a technique to produce cloned offspring of endangered species as well as to investigate nucleus-cytoplasm interactions in mammalian embryo. However, it is still not known which embryo culture medium is optimal for ISCNT embryos for the nuclear donor or the oocyte recipient. We assessed the effects of the culture medium on the developmental competence of the ISCNT embryos by introducing cynomolgus monkey (Macaca fascicularis) fibroblast nuclei into enucleated rabbit (Oryctolagus cuniculus) oocytes (monkey-rabbit embryo). The monkey-rabbit ISCNT embryos that were cultured in mCMRL-1066 developed to the blastocyst stage, although all monkey-rabbit ISCNT embryos cultured in M199 were arrested by the 4-cell stage. When monkey-rabbit ISCNT and rabbit-rabbit somatic cell nuclear transfer (SCNT) embryos were cultured in mCMRL-1066, the blastocyst cell numbers of the monkey-rabbit ISCNT embryos corresponded to the cell numbers of the control rabbit-rabbit SCNT embryos, which were produced from a rabbit fibroblast nucleus and an enucleated rabbit oocyte. In addition, the presence of mitochondria, which were introduced with monkey fibroblasts into rabbit recipient cytoplasm, was confirmed up to the blastocyst stage by polymerase chain reaction (PCR). This study demonstrated that: (1) rabbit oocytes can reprogramme cynomolgus monkey somatic cell nuclei, and support preimplantation development; (2) monkey-rabbit ISCNT embryos developed well in monkey culture medium at early embryonic developmental stages; (3) the cell number of monkey-rabbit ISCNT embryos is similar to that of rabbit-rabbit SCNT embryos; and (4) the mitochondrial fate of monkey-rabbit ISCNT embryos is heteroplasmic from the time just after injection to the blastocyst stage that has roots in both rabbit oocytes and monkey fibroblasts.
  • Yuta Ishizuka; Manami Nishimura; Kazuya Matsumoto; Minoru Miyashita; Toru Takeo; Naomi Nakagata; Yoshihiko Hosoi; Masayuki Anzai
    Theriogenology 80 (5) 421 - 426 2013/09 [Refereed]
  • Ah Reum Lee; Satoshi Kishigami; Tomoko Amano; Kazuya Matsumoto; Teruhiko Wakayama; Yoshihiko Hosoi
    JOURNAL OF REPRODUCTION AND DEVELOPMENT SOCIETY REPRODUCTION & DEVELOPMENT-SRD 59 (3) 238 - 244 0916-8818 2013/06 [Refereed]
     
    Postovulatory mammalian oocyte developmental potential decreases with aging in vivo and in vitro. Aging oocytes typically show cellular fragmentation and chromosome scattering with an abnormally shaped spindle over time. Previously, it was shown that histone acetylation in the mouse oocyte increased during aging and that treatment with trichostatin A (TSA), an inhibitor for class I and II histone deacetylases (HDACs), enhanced the acetylation, that is, aging. In this study, we examined the effect of nicotinamide (NAM), an inhibitor for class III HDACs, on in vitro aging of mouse oocytes as well as TSA. We found that treatment with NAM significantly inhibited cellular fragmentation, spindle elongation and astral microtubules up to 48 h of culture. Although presence of TSA partially inhibited cellular fragmentation and spindle elongation up to 36 h of culture, treatment with TSA induced chromosome scattering at 24 h of culture and more severe cellular fragmentation at 48 h of culture. Further, we found that a-tubulin, a nonhistone protein, increased acetylation during aging, suggesting that not only histone but nonhistone protein acetylation may also increase with oocyte aging. Thus, these data indicate that protein acetylation is abnormally regulated in aging oocytes, which are associated with a variety of aging phenotypes, and that class I/II and class III HDACs may play distinct roles in aging oocytes.
  • Satoshi Nishikawa; Yuki Hatanaka; Mikiko Tokoro; Seung-Wook Shin; Natsumi Shimizu; Takuji Nishihara; Rie Kato; Atsushi Takemoto; Tomoko Amano; Masayuki Anzai; Satoshi Kishigami; Yoshihiko Hosoi; Kazuya Matsumoto
    JOURNAL OF REPRODUCTION AND DEVELOPMENT SOCIETY REPRODUCTION & DEVELOPMENT-SRD 59 (3) 258 - 265 0916-8818 2013/06 [Refereed]
     
    Degradation of maternally stored mRNAs after fertilization is an essential process for mammalian embryogenesis. Maternal mRNA degradation depending on deadenylases in mammalian early embryos has been mostly speculated, rather than directly demonstrated. Previously, we found that gene expression of nocturnin, which functions as a circadian clock-controlled deadenylase in mammalian cells, was clearly changed during the maternal-to-zygotic transition (MZT). Here, we investigated the possible role of nocturnin during mouse MZT. First, we examined the expression profile and localization of nocturnin in mouse oocytes and early embryos. The abundance of Nocturnin mRNA level was significantly decreased from the MII to 4-cell stages and slightly increased from the 8-cell to blastocyst stages, whereas the Nocturnin protein level was almost stable in all examined cells including GV and MII oocytes and early embryos. Nocturnin was localized in both the cytoplasm and the nucleus of all examined cells. We then examined the effect of loss or gain of Nocturnin function on early embryonic development. Knockdown of Nocturnin by injection of Nocturnin antisense expression vector into 1-cell embryos resulted in the delay of early embryonic development to the early blastocyst stage. Moreover, Nocturnin-overexpressed embryos by injection of Nocturnin expression vector impaired their development from the 1-cell to 2-cell or 4-cell stages. These results suggest that precise expression of nocturnin is critical to proper development of early mouse embryos. Functional analysis of nocturnin may contribute to the understanding of the possible role of the deadenylase at mouse MZT.
  • Yuki Hatanaka; Natsumi Shimizu; Satoshi Nishikawa; Mikiko Tokoro; Seung-Wook Shin; Takuji Nishihara; Tomoko Amano; Masayuki Anzai; Hiromi Kato; Tasuku Mitani; Yoshihiko Hosoi; Satoshi Kishigami; Kazuya Matsumoto
    PLOS ONE PUBLIC LIBRARY SCIENCE 8 (4) e60205  1932-6203 2013/04 [Refereed]
     
    After fertilization, the sperm and oocyte genomes undergo extensive epigenetic reprogramming to form a totipotent zygote. The dynamic epigenetic changes during early embryo development primarily involve DNA methylation and demethylation. We have previously identified Gse (gonad-specific expression gene) to be expressed specifically in germ cells and early embryos. Its encoded protein GSE is predominantly localized in the nuclei of cells from the zygote to blastocyst stages, suggesting possible roles in the epigenetic changes occurring during early embryo development. Here, we report the involvement of GSE in epigenetic reprogramming of the paternal genome during mouse zygote development. Preferential binding of GSE to the paternal chromatin was observed from pronuclear stage 2 (PN2) onward. A knockdown of GSE by antisense RNA in oocytes produced no apparent effect on the first and second cell cycles in preimplantation embryos, but caused a significant reduction in the loss of 5-methylcytosine (5 mC) and the accumulation of 5-hydroxymethylcytosine (5 hmC) in the paternal pronucleus. Furthermore, DNA methylation levels in CpG sites of LINE1 transposable elements, Lemd1, Nanog and the upstream regulatory region of the Oct4 (also known as Pou5f1) gene were clearly increased in GSE-knockdown zygotes at mid-pronuclear stages (PN3-4), but the imprinted H19-differential methylated region was not affected. Importantly, DNA immunoprecipitation of 5 mC and 5 hmC also indicates that knockdown of GSE in zygotes resulted in a significant reduction of the conversion of 5 mC to 5 hmC on LINE1. Therefore, our results suggest an important role of maternal GSE for mediating active DNA demethylation in the zygote.
  • Keigo Matsubara; Ah Reum Lee; Satoshi Kishigami; Akio Ito; Kazuya Matsumoto; Hongfang Chi; Norikazu Nishino; Minoru Yoshida; Yoshihiko Hosoi
    BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS ACADEMIC PRESS INC ELSEVIER SCIENCE 434 (1) 1 - 7 0006-291X 2013/04 [Refereed]
     
    Previous studies show that treatment of zygotes with trichostatin A (TSA), a histone deacetylase inhibitor (HDACi), impacts the subsequent development to a blastocyst as well as full-term development. To reveal the dynamics of protein acetylation, with and without TSA treatment during one-cell stage, we examined oocytes and zygotes by immunofluorescence and Western Blot analyses using anti-acetylated lysine and acetylated alpha-tubulin antibodies. In unfertilized oocytes, lysine acetylation level was extremely low over all but faintly detected in the spindle. Once oocyte activation occurs, a dramatic increase of lysine acetylation signal was observed mostly in the pronuclei and a fiber-like structure, the so called midbody, suggesting activation coupled up-regulation of lysine acetylation presumably in histones and alpha-tubulin. TSA treatment resulted in significantly more hyperacetylation not only in the midbody structure and pronuclei but also in the whole cytoplasm. Consistently, Western Blot analysis revealed that acetylation of proteins about 53 kDa and 11 kDa in size, corresponding to alpha-tubulin and histone H4 sizes respectively, were increased mainly after oocyte activation and exclusively enhanced by TSA treatment in zygotes. To confirm this behavior of acetylated nonhistone proteins, acetylated alpha-tubulin was examined and found to be faintly detected in the spindle of MII oocytes but later in whole in the cell of zygotes including the midbody, which was enhanced by TSA treatment. To elucidate the mechanism underlying up-regulation of lysine acetylation following oocyte activation, we assayed the HDAC activity, and found significant reduction of HDAC activity from MII to zygotic stages. Taken together, our data indicate that HDACs play an important role in maintaining low acetylated status in a MII oocyte. However, once an oocyte has been activated, histone and nonhistone proteins including alpha-tubulin are hyperacetylated partly due to a reduction of HDAC activity. TSA treatment of zygotes enhances their acetylation, which could affect subsequent embryonic development. (C) 2013 Elsevier Inc. All rights reserved.
  • Seung-Wook Shin; Natsumi Shimizu; Mikiko Tokoro; Satoshi Nishikawa; Yuki Hatanaka; Masayuki Anzai; Jun Hamazaki; Satoshi Kishigami; Kazuhiro Saeki; Yoshihiko Hosoi; Akira Iritani; Shigeo Murata; Kazuya Matsumoto
    BIOLOGY OPEN COMPANY OF BIOLOGISTS LTD 2 (2) 170 - 182 2046-6390 2013/02 [Refereed]
     
    During the maternal-to-zygotic transition (MZT), maternal proteins in oocytes are degraded by the ubiquitin-proteasome system (UPS), and new proteins are synthesized from the zygotic genome. However, the specific mechanisms underlying the UPS at the MZT are not well understood. We identified a molecule named zygote-specific proteasome assembly chaperone (ZPAC) that is specifically expressed in mouse gonads, and expression of ZPAC was transiently increased at the mouse MZT. ZPAC formed a complex with Ump1 and associated with precursor forms of 20S proteasomes. Transcription of ZPAC genes was also under the control of an autoregulatory feedback mechanism for the compensation of reduced proteasome activity similar to Ump1 and 20S proteasome subunit gene expression. Knockdown of ZPAC in early embryos caused a significant reduction of proteasome activity and decrease in Ump1 and mature proteasomes, leading to accumulation of proteins that need to be degraded at the MZT and early developmental arrest. Therefore, a unique proteasome assembly pathway mediated by ZPAC is important for progression of the mouse MZT. (C) 2012. Published by The Company of Biologists Ltd.
  • 内堀 翔; 清水 なつみ; 畑中 勇輝; 西原 卓志; 武本 淳史; 樋口 智香; 守田 昂太郎; 永井 宏平; 天野 朋子; 岸上 哲士; 細井 美彦; 松本 和也
    日本繁殖生物学会 講演要旨集 日本繁殖生物学会 106 P - 84-P-84 2013 
    【目的】母性遺伝子由来転写産物やタンパク質は,卵胞発育,受精及び胚発生に重要な役割を果たしている。これまでの研究から,母性遺伝子の機能解析において糖タンパク質の一つであるZp3のプロモーターが用いられている(Millar et al., 1991; Linang et al., 1997)。しかし,原始卵胞ではこのプロモーターの転写活性が認められず,また卵母細胞で発現する他のプロモーターについても情報は少ない。我々は,卵母細胞における母性遺伝子の機能解析を行うためのツールとしてHistone H1foo(H1oo)のプロモーター領域を単離し(Tsunemoto et al., 2008),下流にホタルルシフェラーゼ遺伝子を結合した導入遺伝子を組み込んだトランスジェニックマウスを3系統(H14・H16・H19系統)作出した。本実験では,これらのトランスジェニックマウスにおける導入遺伝子の発現を明らかにすることを目的にトランスジェニックマウス由来卵母細胞及び初期胚,各組織におけるルシフェラーゼの発現を検討した。【方法】導入遺伝子が導入された雌マウスに過排卵処置を行い体外受精を施した。培養後,各発生段階でシングルフォトンイメージング装置を用いてルシフェラーゼ活性を測定した。また,各組織をタンパク質抽出し,ルミノメーターを用いて発光量を測定した。【結果及び考察】H16系統での卵母細胞及び初期胚で導入遺伝子の発現が認められ,H1ooの転写制御下でマーカー遺伝子であるホタルルシフェラーゼが発現していることが明らかになった。
  • Seung-Wook Shin; Shigeo Murata; Kazuya Matsumoto
    Journal of Mammalian Ova Research 30 (3) 79 - 85 1341-7738 2013 
    During the maternal-to-zygotic transition (MZT), maternal proteins in oocytes are degraded by the ubiquitin-proteasome system (UPS), which is first event after fertilization, and new proteins are then synthesized from the zygotic genome. Although degradation of accumulated maternal protein is essential for normal early embryonic development, the specific mechanisms underlying the UPS at the MZT are not well understood. We recently provided evidence that proteasomal degradation of maternal proteins is important for the onset of zygotic gene activation (ZGA), and that the zygotespecific proteasome assembly chaperone (ZPAC) plays an important role in the degradation of maternal proteins during mouse MZT. Here, we review why the degradation of maternal proteins via UPS is essential for embryonic reprogramming of the oocyte into a totipotent zygote that is makes somatic development possible. ©2013 Japanese Society of Mammalian Ova Research.
  • H. Sugimoto; Y. Kida; Y. Miyamoto; K. Kitada; K. Matsumoto; K. Saeki; T. Taniguchi; Y. Hosoi
    THERIOGENOLOGY ELSEVIER SCIENCE INC 78 (5) 1040 - 1047 0093-691X 2012/09 [Refereed]
     
    The objective was to develop a culture system that produced blastocyst stage embryos from rabbit oocytes grown in vitro. Two experiments were performed. First, various concentrations of fetal bovine serum (FBS, 0, 0.05, 0.5 and 5%) were used in the culture medium for in vitro growth (IVG) of oocytes recovered from follicles 200 to 299 gm in diameter. Intracytoplasmic sperm injection (ICSI) was performed on mature oocytes obtained after IVG for 8 days and in vitro maturation for 14 to 16 h. Rates of survival and pronuclear formation after ICSI were higher for oocytes grown in a medium with 0.05% FBS compared to oocytes grown in a medium lacking FBS (97.6 vs. 76.9%, 97.5 vs. 70%, P < 0.1). The rate of development to the blastocyst stage was also higher in the medium containing 0.05% FBS than in the medium lacking FBS (9.5 vs. 17.9%, P < 0.05). Next, using oocytes recovered from follicles 200 to 399 gm in diameter which were cultured in 0.05% FBS, oxygen consumption and the number of cells were analyzed. Blastocysts from oocytes grown in vitro with 0.05% FBS had reduced oxygen consumption and number of cells compared with those from ovulated oocytes (21.66 +/- 4.54 X 10(14) vs. 50.19 4.61 X 10(14) mol/sec, 244 +/- 25 vs. 398 +/- 24, P < 0.05). Rabbit oocytes grown in vitro with 0.05% FBS achieved pregnancy, but pregnancies were not maintained to term. In conclusion, the addition of 0.05% PBS to the culture medium for IVG improved developmental competence of rabbit oocytes grown in vitro. (c) 2012 Elsevier Inc. All rights reserved.
  • 池上春香; 小林直彦; 松橋珠子; 武本淳史; 吉廣卓哉; 井上悦子; 加藤里恵; 加藤博己; 田口善智; 天野朋子; 森本康一; 中川優; 入谷明; 松本和也
    日本畜産学会報 83 (3) 281 - 290 1346-907X 2012/08 [Refereed]
  • 池上春香; 小林直彦; 松橋珠子; 武本淳史; 吉廣卓哉; 井上悦子; 加藤里恵; 加藤博己; 田口善智; 天野朋子; 森本康一; 中川優; 入谷明; 松本和也
    日本畜産学会報 83 (3) 281 - 290 1346-907X 2012/08
  • Daisaku Iwamoto; Aya Kasamatsu; Atsushi Ideta; Manami Urakawa; Kazuya Matsumoto; Yoshihiko Hosoi; Akira Iritani; Yoshito Aoyagi; Kazuhiro Saeki
    CELLULAR REPROGRAMMING MARY ANN LIEBERT INC 14 (1) 20 - 28 2152-4971 2012/02 [Refereed]
     
    The success rate of bovine somatic cell nuclear transfer (SCNT) embryos to full term has been reported to be higher with Cl cells than with GO cells. To better understand the reason for this, we analyzed the kinetics of luminescence activity in bovine SCNT embryos from GO and Cl cells carrying a luciferase gene under the control of the beta-actin promoter during early embryonic development. At 60-h postfusion, when bovine embryonic gene activation (EGA) begins, the luminescence activity was higher in G1-SCNT embryos than GO-SCNT embryos. Moreover, half of the G1-SCNT embryos exhibited homogeneous luminescence among the blastomeres, whereas more than half of the GO-SCNT embryos exhibited mosaic luminescence. To characterize the differential luminescence pattern in SCNT embryos, the expressions of several endogenous genes and the level of DNA methylation were determined in all blastomeres of SCNT embryos with or without luminescence. The expressions of several development-related genes (H2AFZ, GJA1, and BAX) and level of DNA methylation of the SCNT embryos with luminescence were the same as those of normal embryos produced by in vitro fertilization. A higher success rate in G1-SCNT embryos is thought to contribute to homogeneous expression among all biastomeres at EGA.
  • Takemoto Atsushi; Nagai Kouhei; Ikegami Haruka; Higuchi Chika; Morita Koutaro; Kobayashi Eiji; Matsumoto Kazuya
    Abstracts for Annual Meeting of Japanese Proteomics Society Japanese Proteomics Society (Japan Human Proteome Organisation) 2012 144 - 144 2012
  • TOMOKO Amano; RIPPERGER Juergen; ALBRECHT Urs; HORIKE Kouta; MATSUMOTO Kazuya
    The Journal of Reproduction and Development Supplement THE SOCIETY FOR REPRODUCTION AND DEVELOPMENT 105 1092 - 1092 2012 
    【目的】 時計遺伝子Per1, Per2は24時間周期の転写量の増減を示し、転写制御因子として多くの遺伝子の転写をリズミカルに制御する。近年、卵巣にも時計遺伝子が周期的に発現することが確認されたが、その卵巣機能への関与は明らかではない。本検討では、野生型マウスとPer1/Per2欠損マウスの雌性生殖生理(排卵、性周期、及びLHサージの状態)を観察し、さらに卵巣において時計遺伝子に転写が制御される遺伝子を探索することにより、卵巣機能への時計遺伝子の関与を検討した。【材料と方法】マウスの排卵と性周期の検討は、膣スメア観察と発情後期の卵管膨大部における排卵卵子の確認によって行った。LHサージの状態は、発情前期における血清中LH濃度の測定(ELISA法)により確認した。また卵巣において時計遺伝子に転写が制御される遺伝子の探索にはマイクロアレイ法を用いた。【結果及び考察】 野生型マウスに比べ、Per1/Per2欠損マウスの排卵の頻度は有意に減少しており(91% vs 20%)、その性周期の回帰の長さは有意に延長していた(5.1 ± 0.1 vs 6.3 ± 0.2 days)。これらの結果は、卵巣にて時計遺伝子が卵巣機能に関与する可能性を示唆したが、同時にPer1/Per2欠損マウスのLHサージが異常であった可能性も示した。そこでPer1/Per2欠損マウスのLHサージの状態(ピーク時の血清中の濃度とタイミング)を野生型マウスと比較したが、大きな差は認められなかった。一方、野生型マウス卵巣とPer1/Per2欠損マウス卵巣に発現する遺伝子をマイクロアレイ法にて解析し、比較を行ったところ、野生型マウスに比べPer1/Per2欠損マウスにて発現量が有意に異なり、卵胞の発育やステロイドホルモンの合成への関与によって、排卵の頻度や性周期に影響すると報告のある27の遺伝子が検出された。これらの結果から、時計遺伝子が卵巣機能に関与する可能性が示された。
  • Ikegami Haruka; Matsuhashi Tamako; Takemoto Atsushi; Nagai Kouhei; Higuchi Tomoka; Morita Koutaro; Kobayashi Naohiko; Matsumoto Kazuya
    Abstracts for Annual Meeting of Japanese Proteomics Society Japanese Proteomics Society (Japan Human Proteome Organisation) 2012 93 - 93 2012
  • LEE Ah Reum; KISHIGAMI Satoshi; MATSUMOTO Kazuya; HOSOI Yoshihiko
    The Journal of Reproduction and Development Supplement THE SOCIETY FOR REPRODUCTION AND DEVELOPMENT 105 1016 - 1016 2012 
    Oocytes have a limited amount of time to support fertilization after ovulation for normal development, for 8-12 h in mouse (Austin, 1974). Unless fertilization occurs within that time, oocytes start to age with meiotic errors such as aberrations in the spindle and misalignment of chromosomes, as well as cellular fragmentation (David L. Keefe, 2004). To elucidate how nuclei contribute to oocyte aging, we examined how reconstructed oocytes by somatic-cell nuclear transfer (SCNT) aged as well as enucleated oocytes. The control and SCNT oocytes started to show cellular fragmentation at 24 h after collection and at 48 h most oocytes were fragmented in the cytoplasm (67/83 = 81%). However, most enucleated oocytes were able to keep a normal morphology, even at 48 h (83/90 = 92%). Recently, we found that acetylation of α-tubulin increases with oocyte aging (A.R. Lee. The 103th meeting of the SRD). Next we examined the acetylated α-tubulin (AC α-tubulin) and α-tubulin in reconstructed aging oocytes. The control, reconstructed and enucleated oocytes during aging gradually increased AC α-tubulin and α-tubulin at 24 h and decreased until 36 h. Furthermore, aster-like structures in the cytoplasm, a symptom of aging, were detected in the control and enucleated aging oocytes at 36 h. The enucleated oocytes started to show aster-like structure earlier than the control in aging oocytes. In contrast, SCNT aging oocytes did not show any aster-like structures in the cytoplasm even at 36 h. Instead, SCNT aging oocytes displayed severe scattered chromosomes and an irregularly shaped spindle. These results provide evidence that the nucleus contributes to phenotypes of the nucleus and cytoplasm in aging oocytes.
  • H. Kato; R. Kitamura; H. Yamaguchi; Y. Numata; T. Kijima; M. Anzai; T. Mitani; K. Matsumoto; K. Saeki; Y. Hosoi; A. Iritani
    REPRODUCTION FERTILITY AND DEVELOPMENT CSIRO PUBLISHING 24 (1) 202 - 202 1031-3613 2012 [Refereed]
  • Etsuko Inoue; Sho Murakami; Takatoshi Fujiki; Takuya Yoshihiro; Atsushi Takemoto; Haruka Ikegami; Kazuya Matsumoto; Masaru Nakagawa
    IPSJ Transactions on Bioinformatics 5 34 - 43 1882-6679 2012 [Refereed]
     
    In this study, we propose a new method to predict three-way interactions among proteins based on correlation coefficient of protein expression profiles. Although three-way interactions have not been studied well, this kind of interactions are important to understand the system of life. Previous studies reported the three-way interactions that based on switching mechanisms, in which a property or an expression level of a protein switches the mechanism of interactions between other two proteins. In this paper, we proposed a new method to predict three-way interactions based on the model in which A and B work together to effect on the expression level of C. We present the algorithm to predict the combinations of three proteins that have the three-way interaction, and evaluate it using our real proteome data.
  • Kazuhiro Saeki; Nobuhiro Kato; Daisaku Iwamoto; Junki Sho; Masao Kishi; Kazuya Matsumoto; Yoshihiko Hosoi; Akira Iritani
    BIOLOGY OF REPRODUCTION SOC STUDY REPRODUCTION 85 0006-3363 2011/07 [Refereed]
  • Kei Miyamoto; Kouhei Nagai; Naoya Kitamura; Tomoaki Nishikawa; Haruka Ikegami; Nguyen T. Binh; Satoshi Tsukamoto; Mai Matsumoto; Tomoyuki Tsukiyama; Naojiro Minami; Masayasu Yamada; Hiroyoshi Ariga; Masashi Miyake; Tatsuo Kawarasaki; Kazuya Matsumoto; Hiroshi Imai
    PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA NATL ACAD SCIENCES 108 (17) 7040 - 7045 0027-8424 2011/04 [Refereed]
     
    Nuclear reprogramming of differentiated cells can be induced by oocyte factors. Despite numerous attempts, these factors and mechanisms responsible for successful reprogramming remain elusive. Here, we identify one such factor, necessary for the development of nuclear transfer embryos, using porcine oocyte extracts in which some reprogramming events are recapitulated. After incubating somatic nuclei in oocyte extracts from the metaphase II stage, the oocyte proteins that were specifically and abundantly incorporated into the nuclei were identified by mass spectrometry. Among 25 identified proteins, we especially focused on a multifunctional protein, DJ-1. DJ-1 is present at a high concentration in oocytes from the germinal vesicle stage until embryos at the four-cell stage. Inhibition of DJ-1 function compromises the development of nuclear transfer embryos but not that of fertilized embryos. Microarray analysis of nuclear transfer embryos in which DJ-1 function is inhibited shows perturbed expression of P53 pathway components. In addition, embryonic arrest of nuclear transfer embryos injected with anti-DJ-1 antibody is rescued by P53 inhibition. We conclude that DJ-1 is an oocyte factor that is required for development of nuclear transfer embryos. This study presents a means for identifying natural reprogramming factors in mammalian oocytes and a unique insight into the mechanisms underlying reprogramming by nuclear transfer.
  • Mikiko Tokoro; Seung-Wook Shin; Satoshi Nishikawa; Hyang-Heun Lee; Yuki Hatanaka; Tomoko Amano; Tasuku Mitani; Hiromi Kato; Masayuki Anzai; Satoshi Kishigami; Kazuhiro Saeki; Yoshihiko Hosoi; Akira Iritani; Kazuya Matsumoto
    JOURNAL OF REPRODUCTION AND DEVELOPMENT SOCIETY REPRODUCTION & DEVELOPMENT-SRD 56 (6) 607 - 615 0916-8818 2010/12 [Refereed]
     
    We investigated the contribution of phosphorylated RNA polymerase II (RNAP II) and dynamic epigenetic changes to the onset of minor zygotic gene activation (ZGA) Using immunofluorescence staining, we observed that the nuclear Localization of RNAP IT was initiated by 6 hours post insemination (hpi), whereas RNAP II phosphorylated at serine residue 5 of the carboxyl-terminal domain (CTD) was localized by 9 hpi, and then RNAP II phosphorylated at serine residue 2 of the CTD was localized in the nucleus of embryos by 12 hpi In a transient gene expression assay using a plasmid reporter gene (p beta-actin/luciferase+/SV40) injected during 6-9 hpi into the male pronucleus, the luciferase+gene was actively transcribed and translated by 13 and 15 hpi, respectively, indicating that a transcriptionally silent state persisted for it least 4 hours after injection We found that the methylation status in the chicken beta-actin promoter region of the plasmid reporter gene may not be associated with the transcriptionally silent state before minor ZGA Exposure to trichostatin A did not induce premature expression of the silent reporter gene injected into 1-cell embryos containing histone deacetylase activity and did not affect the amount of luciferase produced per embryo Acetylated histone H3 lysine 9/14 and acetylated histone H4 lysine 12 and 16 were enriched preferentially in the injected reporter gene at least until 13 hpi, which coincided with the transcriptionally active state Taken together, these results suggest that deposition 01 selectively acetylated histones onto the chromatin of 1-cell embryos functions together with transcriptional elongation by RNAP II and that this sequential chromatin remodeling is involved in the molecular mechanism associated with the onset of minor ZGA in the preimplantation mouse embryo
  • Seung Wook Shin; Mikiko Tokoro; Satoshi Nishikawa; Hyang-Heun Lee; Yuki Hatanaka; Takuji Nishihara; Tomoko Amano; Masayuki Anzai; Hiromi Kato; Tasuku Mitani; Satoshi Kishigami; Kazuhiro Saeki; Yoshihiko Hosoi; Akira Iritani; Kazuya Matsumoto
    JOURNAL OF REPRODUCTION AND DEVELOPMENT SOCIETY REPRODUCTION & DEVELOPMENT-SRD 56 (6) 655 - 663 0916-8818 2010/12 [Refereed]
     
    In mammalian oocytes, the ubiquitin-proteasome system (UPS) is suggested to play important roles in oocyte meiosis resumption, spindle assembly, polar body emission and pronuclear formation by regulating cyclin B1 degradation Ho Never, little is known about the direct relationship between zygotic gene activation (ZGA) and degradation of maternal proteins Here, we investigated the role of the UPS in the onset of ZGA in early mouse embryo First, we found degradation of cyclin B1 protein in fertilized oocytes at 1 hpi by western blot analysis and used the se oocytes throughout this study Subsequently, we determined optimal experimental conditions for transient inhibition of proteasomal activity by specific and reversible proteasomal inhibitor MG132 in the Cl phase of the first cell cycle Under the selected optimal conditions, we subjected transient MG132-treated embryos to reverse transcription (RT)-PCR analysis of expression of four ZGA genes, i e, the hsp70 1, MuERV-L, eif-1a and zscan4d genes As a result, we found that onset of expression of the four examined ZGA genes was delayed m both normally developed 2-cell embryos and arrested 1-cell embryos Our results indicate that proteasomal degradation of proteins by the UPS plays a pivotal role in the molecular mechanisms of ZGA in early mouse embryos
  • Tomoko Amano; Kaori Tokunaga; Reiko Kakegawa; Ayaka Yanagisawa; Atsushi Takemoto; Atsuhiro Tatemizo; Tatsuya Watanabe; Yuki Hatanaka; Akinori Matsushita; Masao Kishi; Masayuki Anzai; Hiromi Kato; Tasuku Mitani; Satoshi Kishigami; Kazuhiro Saeki; Yoshihiko Hosoi; Akira Iritani; Kazuya Matsumoto
    ANIMAL REPRODUCTION SCIENCE ELSEVIER SCIENCE BV 121 (3-4) 225 - 235 0378-4320 2010/09 [Refereed]
     
    We previously showed that circadian genes clock, bmal1, cry1, cry2, per1, and per2 are expressed and function as maternal mRNA regulating events in the oocytes and preimplantation embryos of mice. Recent evidence indicates however that either or both expression profiles of circadian genes in some tissues, and transcript sequences of circadian genes, differ to generate the physiological differences between diurnal and nocturnal species. We therefore investigated the expression profiles of circadian genes in oocytes and preimplantation embryos of species other than mice, namely cattle and rabbits, representing diurnal and nocturnal species, respectively, and determined the protein sequences of circadian genes in these species. Quantitative real-time PCR revealed that all circadian genes considered in this study were present in the oocytes and preimplantation embryos of both species, and the transcript amounts of clock, cry1 and per1 contained in oocytes were significantly higher than in preimplantation embryos of both species. The transcripts of clock, cry1, and per1 of cattle and rabbits were determined by primer walking, and functional domains in the estimated amino acid sequences were compared between cattle and rabbits and with those of humans and mice. The sequences of clock, cry1, and per1 in cattle and rabbits closely resembled those in mice (85-100% homologies), and no difference based on diurnality or nocturnality was observed. These findings suggest that circadian genes in the oocytes and preimplantation embryos of mammals fulfill the same functions across species as maternal mRNA. (C) 2010 Elsevier B.V. All rights reserved.
  • 天野 朋子; 入谷 明; 松本 和也
    Japanese Society for Chronobiology 16 31 - 41 2010
  • Toshiyuki Takehara; Takeshi Teramura; Yuta Onodera; Satoshi Kishigami; Kazuya Matsumoto; Kazuhiro Saeki; Kanji Fukuda; Yoshihiko Hosoi
    STEM CELLS AND DEVELOPMENT MARY ANN LIEBERT, INC 18 (10) 1433 - 1440 1547-3287 2009/12 [Refereed]
     
    Neural stem cells (NSCs) are tissue-specific stem cells with self-renewal potential in brain, and are committed cells of the central nervous system. Recently, some reports have suggested the possibility of the NSCs to differentiate into non-CNS mesodermal derivatives, such as blood cells and skeletal muscle cells. Here we isolated NSCs as neurospheres from a neonatal mouse brain using serum replacement medium, and demonstrated that the stem cell population expressing pluripotent-related genes such as Oct-4, Sox-2, and Nanog possess multiple differentiation potentials to ectodermal, mesodermal, and endodermal lineages, that is, some neural cells, beating cardiomyocytes, adipocytes, and insulin-producing cells. The results of the present study partly provide further evidence for multiple differentiation properties of NSCs and suggest common characteristics between NSCs and other pluripotent stem cells.
  • Shuntaro Ikeda; Atsuhiro Tatemizo; Daisaku Iwamoto; Shunji Taniguchi; Yoichiro Hoshino; Tomoko Amano; Kazuya Matsumoto; Yoshihiko Hosoi; Akira Iritani; Kazuhiro Saeki
    ZYGOTE CAMBRIDGE UNIV PRESS 17 (3) 209 - 215 0967-1994 2009/08 [Refereed]
     
    Histone acetylation is one of the major mechanisms of epigenetic reprogramming of gamete genomes after fertilization to establish a totipotent state for normal development. In the present study, the effects of trichostatin A (TSA), an inhibitor of histone deacetylase, during in vitro fertilization (IVF) of bovine oocytes on subsequent embryonic development were investigated. Cumulus-enclosed oocytes obtained from slaughterhouse bovine ovaries were matured in vitro and subjected to IVF in a defined medium supplemented with 0 (control), 5, 50, and 500 nM TSA for 18 h. After IVF, presumptive zygotes were cultured in modified synthetic oviductal fluid (mSOF) medium until 168 h postinsemination (hpi). Some oocytes were immunostained using antibody specific for histone H4-acetylated lysine 5 at 10 hpi. Cleavage, blastocyst development and cell number of inner cell mass (ICM) and trophectoderm (TE) of blastocysts were assessed. TSA treatment enhanced histone acetylation that was prominent in decondensed sperm nuclei. TSA did not affect the postfertilization cleavage, blastocyst rates, and TE cell number. However, it significantly enhanced ICM cell number (p < 0.05). These results indicate that TSA treatment during IVF of bovine oocytes does not affect blastocyst development but alters the cell number of ICM, suggesting that overriding epigenetic modification of the genome during fertilization has a carryover effect on cell proliferation and differentiation in preimplantation embryos. Thus, further environmental quality controls in assisted reproductive technologies are needed in terms of factors which affect chromatin remodelling.
  • Manabu Satoh; Mikiko Tokoro; Haruka Ikegami; Kouhei Nagai; Youhei Sono; Seung-Wook Shin; Satoshi Nishikawa; Kazuhiro Saeki; Yoshihiko Hosoi; Akira Iritani; Aisaku Fukuda; Yoshiharu Morimoto; Kazuya Matsumoto
    JOURNAL OF REPRODUCTION AND DEVELOPMENT SOCIETY REPRODUCTION & DEVELOPMENT-SRD 55 (3) 316 - 326 0916-8818 2009/06 [Refereed]
     
    Functional and structural changes in the mammalian ovary are coordinately regulated by the pituitary glycoprotein hormones, follicle-stimulating hormone (FSH) and luteinizing hormone (LH), leading to follicular development, ovulation and transformation of follicles into corpus lutea. To investigate protein profiles during these processes of the mouse ovarian cycle, we applied combined methods (two-dimensional gel electrophoresis [2-DE] for separation and visualization of proteins plus matrix laser desorption/ionization time-of-flight mass spectrometry [MALDI-TOF/MS] analysis for protein identification) for comparative proteomic analysis using immature mice at 3 weeks of age. Protein profiles were obtained from proteins extracted from intact ovaries that had been collected from pregnant mare serum gonadotropin (PMSG)/human chorionic gonadotropin (hCG)-primed immature mice at 0 (no PMSG), 24 and 48 h post PMSG, as well as at 10 and 20 h post hCG. The results showed that 1028 common protein spots were found in representative gels that had been separated in the 3 to 11 pH range and the 15-200 kDa range, 253 protein spots (24.6%) of which were differentially expressed (p<0.05) during the mouse ovarian cycle. Of these 253 protein spots, 99 were identified by MALDI-TOF/MS. This comparative proteomic approach to identifying proteins that were potentially involved in the complex process of the ovarian cycle could contribute to our understanding of the molecular basis of functional and structural changes in the ovary in response to gonadotropins. Furthermore, the interesting ovarian proteins identified in this study may eventually serve as diagnostic biomarker candidates of ovarian function.
  • Takeshi Teramura; Yuta Onodera; Hideki Murakami; Syunsuke Ito; Toshihiro Mihara; Toshiyuki Takehara; Hiromi Kato; Tasuku Mitani; Masayuki Anzai; Kazuya Matsumoto; Kazuhiro Saeki; Kanji Fukuda; Norimasa Sagawa; Yoshihiko Hosoi
    JOURNAL OF REPRODUCTION AND DEVELOPMENT SOCIETY REPRODUCTION & DEVELOPMENT-SRD 55 (3) 283 - 292 0916-8818 2009/06 [Refereed]
     
    The embryos of some rodents and primates can precede early development without the process of fertilization; however, they cease to develop after implantation because of restricted expressions of imprinting genes. Asexually developed embryos are classified into parthenote/gynogenote and androgenote by their genomic origins. Embryonic stem cells (ESCs) derived from asexual origins have also been reported. To date, ESCs derived from parthenogenetic embryos (PgESCs) have been established in some species, including humans, and the possibility to be alternative sources for autologous cell transplantation in regenerative medicine has been proposed. However, some developmental characteristics, which might be important for therapeutic applications, such as multiple differentiation capacity and transplantability of the ESCs of androgenetic origin (AgESCs) are uncertain. Here, we induced differentiation of mouse AgESCs and observed derivation of neural cells, cardiomyocytes and hepatocytes in vitro. Following differentiated embryoid body (EB) transplantation in various mouse strains including the strain of origin, we found that the EBs Could engraft in theoretically MHC-matched strains. Our results indicate that AgESCs possess at least two important characteristics, multiple differentiation properties in vitro and transplantability after differentiation, and suggest that they can also serve as a source of histocompatible, tissues for transplantation.
  • Miyuri Kawasumi; Yuichi Unno; Toshiki Matsuoka; Megumi Nishiwaki; Masayuki Anzai; Tomoko Amano; Tasuku Mitani; Hiromi Kato; Kazuhiro Saeki; Yoshihiko Hosoi; Akira Iritani; Satoshi Kishigami; Kazuya Matsumoto
    MOLECULAR REPRODUCTION AND DEVELOPMENT WILEY-LISS 76 (4) 342 - 350 1040-452X 2009/04 [Refereed]
     
    Oct-4 is essential for normal embryonic development, and abnormal Oct-4 expression in cloned embryos contributes to cloning inefficiency. However, the causes of abnormal Oct-4 expression in cloned embryos are not well understood. As DNA methylation in regulatory regions is known to control transcriptional activity, we investigated the methylation status of three transcriptional regulatory regions of the Oct-4 gene in cloned mouse embryos-the distal enhancer (DE), the proximal enhancer (PE), and the promoter regions. We also investigated the level of Oct-4 gene expression in cloned embryos. Immunochemistry revealed that 85% of cloned blastocysts expressed Oct-4 in both trophectoderm and inner cell mass cells. DNA methylation analysis revealed that the PE region methylation was greater in cloned morulae than in normal morulae. However, the same region was less methylated in cloned blastocysts than in normal blastocysts. We found abnormal expression of de novo methyltransferase 3b in cloned blastocysts. These results indicate that cloned embryos have aberrant DNA methylation in the CpG sites of the PE region of Oct-4, and this may contribute directly to abnormal expression of this gene in cloned embryos.
  • Yoriko Indo; Atsuhiro Tatemizo; Yuki Abe; Iwane Suzuki; Kazuya Matsumoto; Yoshihiko Hosoi; Mikio Kinoshita; Koji Mikami; Norio Murata; Akira Iritani; Kazuhiro Saeki
    Biochimica et Biophysica Acta - Molecular and Cell Biology of Lipids 1791 183 - 190 1388-1981 2009/03 [Refereed]
     
    Long-chain n-3 fatty acids can lower the risk of lifestyle-related diseases, therefore, we introduced a plant fatty acid desaturation3 (FAD3) gene into mammalian cells. The FAD3 cDNA was isolated from the immature seeds of scarlet flax and optimized to human high-frequency codon usage for enhancement of its expression levels in mammalian cells (hFAD3). We introduced the gene into bovine muscle satellite cells, which can be differentiated into multilocular adipocytes in vitro. After hFAD3 transfection, the cells were differentiated into adipocytes and their fatty acid composition was analyzed by gas chromatography. The level of α-linolenic acid (18:3n-3) in transfected adipocytes increased about ten-fold compared with non-transfected adipocytes. In addition, the levels of docosapentaenoic acid (DPA, 22:5n-3) and docosahexaenoic acid (DHA, 22:6n-3) in transfected adipocytes were significantly higher than those in non-transfected adipocytes. Moreover, we produced bovine cloned embryos from the hFAD3 cells by somatic cell nuclear transfer. Blastocyst rates of hFAD3 clones were the same as the control clones using the non-transfected cells (21% vs 27%, P > 0.05). hFAD3 transcripts were detected in all of the blastocysts. These results demonstrate the functional expression of a plant hFAD3 in mammalian adipocytes, and normal development of cloned embryos carrying the hFAD3 gene. © 2009 Elsevier B.V. All rights reserved.
  • Tomoko Amano; Akinori Matsushita; Yuki Hatanaka; Tatsuya Watanabe; Katsutaka Oishi; Norio Ishida; Masayuki Anzai; Tasuku Mitani; Hiromi Kato; Satoshi Kishigami; Kazuhiro Saeki; Yoshihiko Hosoi; Akira Iritani; Kazuya Matsumoto
    BIOLOGY OF REPRODUCTION SOC STUDY REPRODUCTION 80 (3) 473 - 483 0006-3363 2009/03 [Refereed]
     
    In mammals, circadian genes, Clock, Arntl (also known as Bmal1), Cry1, Cry2, Per1, Per2, and Per3, are rhythmically transcribed every 24 h in almost all organs and tissues to tick the circadian clock. However, their expression and function in oocytes and preimplantation embryos have not been investigated. In this study we found that the circadian clock may stop in mouse oocytes and preimplantation embryos. Real-time PCR analysis revealed the presence of transcripts of these genes in both oocytes and preimplantation embryos; however, their amounts did not oscillate every 24 h in one- to four-cell and blastocyst-stage embryos. Moreover, immunofluorescence analyses revealed that CLOCK, ARNTL, and CRY1 were localized similarly in the nuclei of germinal vesicle (GV) oocytes and one-cell- to four-cell-stage embryos. Because CRY1 is known to interact with the CLOCK-ARNTL complex to suppress transcription-promoting activity of the complex for genes such as Wee1, Cry2, Per1, Per2, and Per3 in cells having the ticking circadian clock, we hypothesized that if the circadian clock functions in GV oocytes and one-cell- to four-cell-stage embryos, CLOCK, ARNTL, and CRY1 might suppress the transcription of these genes in GV oocytes and one-cell- to 4-cell-stage embryos as well. As a result, knockdown of CRY1 in GV oocytes by RNA interference did not affect the transcription levels of Wee1, Cry2, Per1, Per2, and Per3, but it reduced maturation ability. Thus, it seems that circadian genes are not involved in circadian clock regulation in mouse oocytes and preimplantation embryos but are involved in physiologies, such as meiosis.
  • Indo Y; Tatemizo A; Abe Y; Suzuki I; Matsumoto K; Hosoi Y; Kinoshita M; Mikami K; Murata N; Iritani A; Saeki K
    Biochimica et biophysica acta 1791 (3) 183 - 190 0006-3002 2009/03 [Refereed]
  • Indo Y; Tatemizo A; Abe Y; Suzuki I; Matsumoto K; Hosoi Y; Kinoshita M; Mikami K; Murata N; Iritani A; Saeki K
    Biochimica et biophysica acta 0006-3002 2009/01 [Refereed]
  • 永井宏平; 吉廣卓哉; 井上悦子; 池上春香; 園陽平; 川路英哉; 小林直彦; 松橋珠子; 大谷健; 森本康一; 中川優; 入谷明; 松本和也
    日本畜産学会報 79 (4) 467 - 481 1346-907X 2008/11 [Refereed]
  • 永井宏平; 吉廣卓哉; 井上悦子; 池上春香; 園陽平; 川路英哉; 小林直彦; 松橋珠子; 大谷健; 森本康一; 中川優; 入谷明; 松本和也
    日本畜産学会報 79 (4) 467 - 481 1346-907X 2008/11
  • Ryo Kakegawa; Takeshi Teramura; Toshiyuki Takehara; Masayuki Anzai; Tasuku Mitani; Kazuya Matsumoto; Kazuhiro Saeki; Norimasa Sagawa; Kanji Fukuda; Yoshihiko Hosoi
    JOURNAL OF REPRODUCTION AND DEVELOPMENT SOCIETY REPRODUCTION & DEVELOPMENT-SRD 54 (5) 352 - 357 0916-8818 2008/10 [Refereed]
     
    Primordial germ cells (PGCs) are embryonic precursors of the gametes of adult animals and are considered stem cells of the germline. Since their proliferation in vitro correlates well with the schedule of developmental changes in vivo, they might be interesting research tools for genomic imprinting, germ-cell tumors and fertility. Furthermore, once primordial germ cells are separated and placed on a feeder layer with cytokines, they become Cultured pluripotent cell lines called embryonic germ (EG) cells. EG cells share several important characteristics with embryonic stem (ES) cells as they call also contribute to the germ line of chimeras. To investigate the characteristics of PGCs and establish rabbit EG (rEG) cells, we cultured rabbit PGCs (rPGCs) in vitro with various combinations of leukemia inhibitory factor (LIF), basic fibroblast growth factor (bFGF) and forskolin oil inactivated mouse embryonic fibroblast (MEF) feeder layers. The present study found PGC proliferation in early cultures and induction of rEG-like colonies. These cells expressed pluripotent markers, such as alkaline phosphatase activity, OCT-4, Sox-2 and SSEA-1, in the undifferentiated state; however, the cells did not develop into a teratoma when injected into the kidney capsules of SCID mice, although the restricted differentiation potentials to neural cells were determined via embryoid body formation. From these characteristics and further characterization of the germ stem cell markers Vasa, SCP-1 and SCP-3, we suggested that these were hybrid cells with characteristics somewhere between PGC and EG cells.
  • Kazunobu Tsunemoto; Masayuki Anzai; Toshiki Matsuoka; Mikiko Tokoro; Seung-Wook Shin; Tomoko Amano; Tasuku Mitani; Hiromi Kato; Yoshihiko Hosoi; Kazuhiro Saeki; Akira Iritani; Kazuya Matsumoto
    MOLECULAR REPRODUCTION AND DEVELOPMENT WILEY-LISS 75 (7) 1104 - 1108 1040-452X 2008/07 [Refereed]
     
    We examined the promoter activities of three mouse maternal genes (H1oo, Npm2, and Zar1) in oocytes and pre-implantation embryos, and examined the promoters for cis-acting elements of 5'-flanking region to obtain the best promoter for inducing oocyte-specific gene expression. For the assay, we injected firefly luciferase gene constructs under the control of the promoters into the oocytes and embryos. Each promoter region showed transcriptional activity in oocytes, but not in fertilized embryos. Deletion analysis showed that a putative E-box region at position -72 of the H1oo promoter and at the -180 of the Npm2 promoter were required for basal transcriptional activity in oocytes. Moreover, a putative NBE motif (NOBOX DNA binding elements) (-1796) was shown to enhance basal transcriptional activity of the Npm2 promoter. Thus, the E-box and/or NBE may be key regulatory regions for the expression of the examined maternal genes (H1oo and Npm2) in growing mouse oocytes.
  • Toshiki Matsuoka; Manabu Sato; Mikiko Tokoro; Seung-Wook Shin; Atsuto Uenoyama; Kazunari Ito; Syuji Hitomi; Tomoko Amano; Masayuki Anzai; Hiromi Kato; Tasuku Mitani; Kazuhiro Saeki; Yoshihiko Hosoi; Akira Iritani; Kazuya Matsumoto
    JOURNAL OF REPRODUCTION AND DEVELOPMENT SOCIETY REPRODUCTION & DEVELOPMENT-SRD 54 (3) 192 - 197 0916-8818 2008/06 [Refereed]
     
    We isolated a mouse cDNA, zag1 (zygotic gene activation-associated gene 1), that has an open reading frame of 1,728-bp encoding a protein of 66.2 kDa including both a bipartite nuclear targeting sequence and a P-loop motif containing nucleoside triphosphate hydrolase motifs. Northern blot analysis of mouse tissues showed that zag1 was widely expressed but was especially prominent in the ovary and testis. RT-PCR analysis of in vitro fertilized embryos showed that the abundance of zag1 transcripts in oocytes decreased after fertilization, and zag1 mRNA was detected at 15 h post insemination (hpi) in fertilized embryos indicating that the gene was expressed at the start of zygotic gene activation at the mouse 1-cell stage. The nuclear-localization of ZAG1 protein in mouse preimplantation embryos at 15 hpi was confirmed by both subcellular analysis of enhanced green fluorescent protein (EGFP)-tagged ZAG1 and immunocytochemical analysis with anti-ZAG1 antibody. Subsequently, using yeast two-hybrid screening, we identified U2 small nuclear ribonucleoprotein B (U2B"), which is associated with pre-mRNA splicing, as a putative interacting partner of ZAG1 protein. Furthermore, knockdown of zag1 expression by an antisense DNA plasmid induced arrest and/or delay of embryonic development in injected 1-cell embryos. These results suggest that ZAG1 may be closely associated with zygotic gene expression in mouse preimplantation embryos.
  • H. Kato; A. Nakao; M. Nishiwaki; M. Anzai; T. Mitani; K. Matsumoto; K. Saeki; Y. Hosoi; A. Iritani
    REPRODUCTION FERTILITY AND DEVELOPMENT CSIRO PUBLISHING 20 (1) 100 - 100 1031-3613 2008 [Refereed]
  • S. Taniguchi; N. Hayashi; Y. Abe; D. Iwamoto; S. Kishigami; M. Kishi; H. Kato; T. Mitani; K. Matsumoto; Y. Hosoi; A. Iritani; Y. Nagao; K. Saeki
    REPRODUCTION FERTILITY AND DEVELOPMENT CSIRO PUBLISHING 20 (1) 110 - 110 1031-3613 2008 [Refereed]
  • D. Iwamoto; S. Kishigami; S. Taniguchi; Y. Abe; T. Matsui; A. Kasamatsu; A. Tatemizo; T. Mitani; H. Kato; K. Matsumoto; Y. Hosoi; T. Wakayama; A. Iritani; K. Saeki
    REPRODUCTION FERTILITY AND DEVELOPMENT CSIRO PUBLISHING 20 (1) 99 - 99 1031-3613 2008 [Refereed]
  • 畑中 勇輝; 天野 朋子; 松本 和也
    Memory of Insitute of Advanved Technology, Kinki University 近畿大学先端技術総合研究所 13 (13) 1 - 9 1346-8693 2008 [Refereed]
     
    概日周期は多くの動物の生理学的現象に観察される。ヒトを始めとする哺乳類においても、概日周期のかく乱は妊孕性の低下を引き起こすなど、健康に密接な関わりがあることが明らかにされている。一方、それぞれの生理学的現象における概日周期形成の分子制御機序を明らかにする研究が進むにつれて、時計遺伝子群による転写制御機構は、概日周期を上流で制御する分子機序であることが認識されるようになっている。最近、時計遺伝子群の制御下にある遺伝子群の中に、細胞周期制御に関係する遺伝子が含まれていることが明らかにされ、動物個体の組織や器官における時計遺伝子群の生理学的役割が徐々に解明されつつある。本稿では、時計遺伝子群による細胞周期制御に関する最近の知見を紹介するとともに、雌個体の生殖器官である卵巣と子宮の生理学的機能に、時計遺伝子群による細胞周期制御が関与する可能性について展望する。
  • Masataka Furuya; Mamoru Tanaka; Takahide Teranishi; Kazuya Matsumoto; Yoshihiko Hosoi; Kazuhiro Saeki; Hitoshi Ishimoto; Kazuhiro Minegishi; Akira Iritani; Yasunori Yoshimura
    JOURNAL OF REPRODUCTION AND DEVELOPMENT SOCIETY REPRODUCTION & DEVELOPMENT-SRD 53 (4) 895 - 902 0916-8818 2007/08 [Refereed]
     
    Oocyte-specific linker histone H1foo is localized in the oocyte nucleus, either diffusely or bound to chromatin, during the processes of meiotic maturation and fertilization. This expression pattern suggests that H1foo plays a key role in the control of gene expression and chromatin modification during oogenesis and early embryogenesis. To reveal the function of H1foo, we microinjected antisense morpholino oligonucleotides (MO) against H1foo into mouse germinal-vesicle stage oocytes. The rate of in vitro maturation of the antisense MO group was significantly lower than that of the control group. Eggs that failed to extrude a first polar body following injection of antisense MO arrested at metaphase I. Additionally, co-injection of in vitro synthesized H1foo mRNA along with antisense MO successfully rescued expression of H1foo and improved the in vitro maturation rate. There was no difference in the rate of parthenogenesis between the antisense MO and control groups. These results indicate that H1foo is essential for maturation of germinal vesicle-stage oocytes.
  • Hiroyuki Kanaya; Shu Hashimoto; Takeshi Teramura; Yoshiharu Morimoto; Kazuya Matsumoto; Kazuhiro Saeki; Akira Iritani; Yoshihiko Hosoi
    JOURNAL OF REPRODUCTION AND DEVELOPMENT SOCIETY REPRODUCTION & DEVELOPMENT-SRD 53 (3) 631 - 637 0916-8818 2007/06 [Refereed]
     
    To clarify the mechanism that impairs development of in vitro grown (IVG) oocytes, we assessed whether the developmental disability of IVG oocytes is caused by cytoplasmic dysfunction. First, we assessed the cleavage of nuclear-substituted oocytes cultured in vitro. The nuclei, but not the cytoplasm, of the IVG oocytes were able to support subsequent cleavage after artificial activation. The mitochondrial activity of the oocytes increased as the follicles grew. However, the mitochondrial activity of the IVG oocytes was significantly lower than that of ovulated oocytes and oocytes recovered from follicles with diameters of more than 300 mu m. Furthermore, the expression levels of mitochondrial transcriptional factor A (TFAM) in the oocytes increased in a similar manner. However, the expression levels of TFAM in the IVG oocytes was significantly lower than that of ovulated oocytes and oocytes recovered from follicles with diameters of more than 300 mu m. Taken together, these results indicate that the low developmental competence of IVG oocytes is caused by a cytoplasm deficiency due to low mitochondrial activity.
  • Miyuri Kawasumi; Masayuki Anzai; Toshiyuki Takehara; Tasuku Mitani; Hiromi Kato; Kazuhiro Saeki; Akira Iritani; Kazuya Matsumoto; Yoshihiko Hosoi
    JOURNAL OF REPRODUCTION AND DEVELOPMENT SOCIETY REPRODUCTION & DEVELOPMENT-SRD 53 (3) 615 - 622 0916-8818 2007/06 [Refereed]
     
    The majority of somatic cell nuclear transferred (SCNT) embryos die before or after implantation. Many studies have focused on morphological remodeling of the donor nucleus and its associated cytoskeletal structures in the early events of nuclear transfer. However, little is known about the 2-cell stage of SCNT embryos after the first division. In this study, we compared the morphological status of chromosomal division during the 1-cell stage to the 2-cell stage in SCNT embryos with that in intracytoplasmic sperm injection (ICSI) embryos. The microtubules and cytoplasmic asters, which are related to chromatin segregation, disappeared at the pronuclear stage, although formation of the first mitotic spindle was normal in both the SCNT and ICSI embryos. However, nuclear fragmentation was observed in 30% of the 2-cell SCNT embryos and 12% of the 2-cell ICSI embryos. Nuclear fragmentation was present in both blastomeres of these embryos. No apoptotic DNA fragmentation was observed in TdT-mediated dUTP-biotin Nick End Labeling (TUNEL) assays for either the SCNT or ICSI embryos. In both the SCNT and ICSI embryos, the distribution of chromosomes in the first mitotic spindle was disturbed during the process of division from the 1-cell stage to the 2-cell stage. These results suggest that loss of SCNT embryos just before or after implantation may be due to an abnormal chromosome distribution at the 2-cell stage.
  • H. Kato; T. Koda; M. Kishimoto; T. Mitani; K. Matsumoto; K. Saeki; Y. Hosoi; A. Iritani
    REPRODUCTION FERTILITY AND DEVELOPMENT CSIRO PUBLISHING 19 (1) 277 - 278 1031-3613 2007 [Refereed]
  • T. Mitani; T. Nagai; T. Masutani; H. Kato; K. Saeki; K. Matsumoto; Y. Hosoi; A. Iritani
    REPRODUCTION FERTILITY AND DEVELOPMENT CSIRO PUBLISHING 19 (1) 230 - 230 1031-3613 2007 [Refereed]
  • Effects of trichostatin A on development of bovine somatic cell nuclear transfer embryos.
    Iwamoto D; Saeki K; Kishigami S; Kasamatsu A; Tatemizo A; Abe Y; Ikeda S; Taniguchi S; Mitani T; Kato H; Matsumoto K; Hosoi Y; Wakayama T; Iritani A
    Reprod. Fertil. Develop. 19 (1) 142  2007/01 [Refereed]
  • DNA methylation profiles of upstream elements of Oct-3/4 gene in in vitro fertilization (IVF) and somatic cell nuclear-transferred (SCNT) embryos.
    Kawasumi M; Unno Y; Nishiwaki M; Matsumoto K; Anzai M; Amano T; Mitani T; Kato H; Saeki K; Hosoi Y; Iritani A
    Reprod. Fertil. Develop. 19 (1) 143  2007/01 [Refereed]
  • Identification and characterization of the 5’-flanking region of three mouse maternal genes (Histone H1oo, Nucleoplasmin2, and Zygote arrest 1) : Transcriptional activity in mouse oocytes.
    Tsunemoto K; Matsumoto K; Anzai M; Hayakumo M; Amano T; Mitani T; Kato H; Hosoi Y; Saeki K; Iritani A
    Reprod. Fertil. Develop. 19 (1) 257 - 258 2007/01 [Refereed]
  • Takeshi Teramura; Toshiyuki Takehara; Nobuyuki Kawata; Nahoko Fujinami; Tasuku Mitani; Makoto Takenoshita; Kazuya Matsumoto; Kazuhiro Saeki; Akira Iritani; Norimasa Sagawa; Yoshihiko Hosoi
    Cloning and Stem Cells 9 (2) 144 - 156 1536-2302 2007 [Refereed]
     
    Embryonic stem cells (ESCs) of nonhuman primates are important for research into human gametogenesis because of similarities between the embryos and fetuses of nonhuman primates and those of humans. Recently, the formation of germ cells from mouse ESCs in vitro has been reported. In this study, we established cynomolgus monkey ES cell lines (cyESCs) and attempted to induce their differentiation into germ cells to obtain further information on the development of primate germ cells by observing the markers specific to germ cells. Three cyESCs were newly established and confirmed to be pluripotent. When the cells are induced to differentiate, the transcripts of Vasa and some meiotic markers were expressed. VASA protein accumulated in differentiated cell clumps and VASA-positive cells gathered in clumps as the number of differentiation days increased. In the later stages, VASA-positive clumps coexpressed OCT-4, suggesting that these cells might correspond to early gonocytes at the postmigration stage. Furthermore, meiosis-specific gene expression was also observed. These results demonstrate that cyESCs can differentiate to developing germ cells such as primordial germ cells (PGCs) or more developed gonocytes in our differentiation systems, and may be a suitable model for studying the mechanisms of primate germ cell development. © Mary Ann Liebert, Inc.
  • Takeshi Teramura; Toshiyuki Takehara; Naoko Kishi; Toshihiro Mihara; Nobuyuki Kawata; Hiroki Takeuchi; Makoto Takenoshita; Kazuya Matsumoto; Kazuhiro Saeki; Akira Iritani; Norimasa Sagawa; Yoshihiko Hosoi
    Cloning and Stem Cells 9 (4) 485 - 494 1536-2302 2007 [Refereed]
     
    Embryonic stem cells (ESCs) are a good material for the study of mammalian development, production of genetically modified animals, and drug discovery because they proliferate infinitely while maintaining a multilinege differentiation potency and a normal karyotype. However, ethical considerations limit the use of human embryos for the establishment of ESCs. Recently, ESCs have been produced from blastomeres divided by biopsy in mice and humans. The method is expected to be less controversial because it does not destroy the embryo. However, no one has yet produced both a pup and an ESC from a single embryo. Here, we describe the production of individual/ESC pairs from each of three embryos out of 20 attempts, and is thus considered efficient. Blastomere-derived ESC could differentiate some types of tissues and contribute to chimera mouse. These results show that each blastomere at two-cell stage possesses pluripotency and separated blastomeres maintain viability to develop to a pup or pluripotent ESC. © 2007 Mary Ann Liebert, Inc.
  • Masayuki Anzai; Megumi Nishiwak; Miho Yanagi; Tatsuyuki Nakashima; Takehito Kaneko; Yoshitomo Taguchi; Mikiko Tokoro; Seung-Wook Shin; Tasuku Mitani; Hiromi Kato; Kazuya Matsumoto; Naomi Nakagata; Akira Iritani
    JOURNAL OF REPRODUCTION AND DEVELOPMENT SOCIETY REPRODUCTION & DEVELOPMENT-SRD 52 (5) 601 - 606 0916-8818 2006/10 [Refereed]
     
    Development of assisted reproductive technologies is necessary to obtain fertilized oocytes in a subfertile transgenic mouse strain. Here, we showed the application of laser-assisted drilling of the zona pellucida to in vitro fertilization of cryopreserved mouse oocytes with sperm from subfertile transgenic mice (C57BL/6N-Tg(UCP/FAD2)U8 strain). After cryopreservation by vitrification, the recovery and survival rates of the zona-drilled mouse oocytes were 97% (97/100) and 94% (91/97), respectively. In vitro fertilization of the cryopreserved zona-drilled mouse oocytes with sperm from the subfertile transgenic mice was greatly facilitated (60%, 55/91) compared to that of the cryopreserved zona-intact mouse oocytes (11%, 81/768). In vitro fertilized embryos that developed to the 2-cell stage were again cryopreserved by vitrification, and after warming they were transferred into recipient females. Subsequently, six viable offspring were delivered, and all were confirmed to be transgenic mice. These results indicate that laser-assisted zona drilling of oocytes combined with cryopreservation by vitrification may be a useful approach for large-scale production of in vitro fertilized embryos for managing transgenic mouse strains with reproductive disabilities such as subfertile sperm.
  • Satoshi Mizuno; Youhei Sono; Toshiki Matsuoka; Kazuya Matsumoto; Kazuhiro Saeki; Yoshihiko Hosoi; Aisaku Fukuda; Yoshiharu Morimoto; Akira Iritani
    JOURNAL OF REPRODUCTION AND DEVELOPMENT SOCIETY REPRODUCTION & DEVELOPMENT-SRD 52 (3) 429 - 438 0916-8818 2006/06 [Refereed]
     
    We previously identified a novel gonad-specific expression gene (Gse) and investigated its expression during gametogenesis in the mouse testis and ovary. In this study, we generated a polyclonal antibody to GSE protein and determined the profiles of the protein's expression in germ cells and preimplantation embryos in detail using immunocytochemical and immunofluorescence staining. In a Western blot analysis, the anti-GSE antibody recognized long and short isoforms (approximately 27.6 kDa and 23.1 kDa) of the protein in the mouse testis and the long isoform in the ovary. In the mouse testis, GSE protein was expressed in spermatocytes I in the pachytene stage, round spermatids, and elongated spermatids. In the mouse ovary, the protein was located in the cytoplasm and nucleus of all oocytes regardless of the stage of the ovarian follicles. In preimplantation embryos from the pronuclear to blastocyst stage, however, GSE protein was mainly detected in the nuclei of cells. At the blastocyst stage, the protein was confirmed to have accumulated in the inner cell mass (ICM), whereas it had mostly disappeared from the trophectoderm (TE). These findings suggest that GSE protein may play a role in the establishment of nuclear totipotency and may be associated with early lineage specification.
  • Saeki K; Tamari T; Kasamatsu A; Iwamoto D; Kameyama S; Tatemizo A; Mitani T; Kato H; Hosoi Y; Matsumoto K; Taniguchi S; Ideta A; Urakawa M; Aoyagi Y; Iritani A
    REPRODUCTION FERTILITY AND DEVELOPMENT CSIRO PUBLISHING 18 (1-2) 142 - 143 1031-3613 2006 [Refereed]
  • 安齋 政幸; 細井 美彦; 佐伯 和弘; 松本 和也; 入谷 明; 糟谷佳恵
    実験動物技術 40 (2) 87 - 90 0387-978X 2005/12 [Refereed]
  • 安齋 政幸; 細井 美彦; 佐伯 和弘; 松本 和也; 入谷 明
    日本胚移植学雑誌 Japanese Journal of embryo Transfer 27 (3) 123 - 131 1341-2965 2005/09 [Refereed]
  • T Amano; T Mori; K Matsumoto; A Iritani; T Watanabe
    THERIOGENOLOGY ELSEVIER SCIENCE INC 64 (2) 261 - 274 0093-691X 2005/07 [Refereed]
     
    At the time of fertilization, release of inositol 1,4,5-trisphosphate (IP3) into the cytoplasm of oocytes is said to be induced by hydrolysis of phosphatidylinositol his phosphate (P12) via activation of phospholipase C and is responsible for the Ca2+ oscillation in oocytes immediately after sperm penetration. On the other hand, cumulus cells have been reported to play an important role in cytoplasmic maturation of mammalian oocytes and to affect embryonic development after fertilization. To obtain more information on the role of cumulus cells in cytoplasmic maturation of oocytes, the effects of cumulus cells on the rise in [Ca2+]i and the rates of activation and development of porcine mature oocytes induced by IP3 injection were investigated. Mature porcine oocytes that had been denuded of their cumulus cells in the early stage of the maturation period had a depressed rise in [Ca2+]i (4.0-6.0) and reduced rates of activation (31.4-36.8%) and development (10.0-24.4%) induced by IP3 injection compared with those of their cumulus-enclosed counterparts (7.3, 69.1% and 43.8%; P < 0.05). The [Ca2+]i rise and the rates of activation and development depressed by the removal of cumulus cells were restored by adding pyruvate to the maturation medium. Furthermore, the IP3 injection-induced depression of [Ca2+]i rise in mature oocytes derived from cumulus-denuded oocytes (Dos) was restored when they were cultured in a medium with pyruvate (3.9-6.3, P < 0.05). Also, mature oocytes from cumulus-oocyte complexes (COCs) cultured in a medium without glucose had a lower rise in [Ca2+]i than that in mature oocytes from COCs cultured with glucose (7.4-6.0, P < 0.05). Cumulus cells supported porcine oocytes during maturation in the rise in [Ca2+]i induced by IP3 and the following activation and development of porcine oocytes after injection of IP3. Moreover, we inferred that a function of cumulus cells is to produce pyruvate by metabolizing glucose and to provide oocytes with pyruvate during maturation, thereby promoting oocyte sensitivity to IP3. (c) 2004 Elsevier Inc. All rights reserved.
  • K Tanemura; A Ogura; C Cheong; H Gotoh; K Matsumoto; E Sato; Y Hayashi; HW Lee; T Kondo
    DEVELOPMENTAL BIOLOGY ACADEMIC PRESS INC ELSEVIER SCIENCE 281 (2) 196 - 207 0012-1606 2005/05 [Refereed]
     
    Chromosomal structure within the nucleus influences various biological processes such as transcription and replication. Telomeres are located at the end of eukaryotic chromosomes and they can be a decisive factor for correct chromosomal positioning. To gain new insight into telomere dynamics, we examined telomere length and positional changes during spermatogenesis using improved fluorescence in situ hybridization (FISH) and in situ telomeric repeat amplification protocols (TRAP) on histological sections. FISH revealed telomere length and chromosome position within nuclei change dynamically. Telomere extension occurred during spermiogenesis. In situ TRAP analysis verified elevated telomerase activity in elongating spermatids. Together, these data show that elongated spermatids have longer telomeres than precursor spermatogenic cells. This observation indicates that telomere elongation in haploid cells occurs after meiosis and in the absence of genomic replication. Analyses of testes from telomerase null mice further support the significance of telomere dynamics during spermatogenesis and the existence of an alternative telomere extension pathway. (C) 2005 Elsevier Inc. All rights reserved.
  • K Saeki; N Sumitomo; Y Nagata; N Kato; Y Hosoi; K Matsumoto; A Iritani
    JOURNAL OF REPRODUCTION AND DEVELOPMENT SOCIETY REPRODUCTION & DEVELOPMENT-SRD 51 (2) 293 - 298 0916-8818 2005/04 [Refereed]
     
    The atomic force microscope (AFM) provides nanometer resolution, topographic data of the natural surface structure of materials. We studied the topology of the surface structure of bovine sperm heads during the acrosome reaction by AFM. In addition, we numerically analyzed the areas of the median sagittal plane of the sperm heads. Bovine frozen-thawed spermatozoa were washed, capacitated by heparin, and incubated with lysophosphaticlylcholine (LPC) to induce the acrosome reaction, smeared on a cover glass, air-dried, and observed with AFM using the dynamic force (tapping) mode. AFM analysis of spermatozoa showed the clear surface structure of acrosomes, equatorial segments, postacrosomal regions and necks. Although AFM images of spermatozoa capacitated by heparin had complete acrosomes, most spermatozoa treated with LPC had no acrosomal caps as shown by AFM. These observations coincided with those obtained by light microscopy after staining with naphthol yellow S and erythrosin B. Furthermore, numerical analysis of AFM images indicated that areas of the median sagittal plane of the anterior portions of acrosomereacted sperm heads (2679 +/- 616 pixels) were approximately 40% less than those of intact heads (4535 +/- 174 pixels, P < 0.05). These results indicate that AFM can usefully observe and numerically analyze the fine surface structures of bovine spermatozoa.
  • Yuichi Unno; Miyuri Kawasumi; Kazuya Matsumoto; Tomoko Amano; Kazuhiro Saeki; Yoshihiko Hosoi; Akira Iritani; Hiromi Kato; Kazuya Matsumoto; Masayuki Anzai; Tasuku Mitani; Kazuhiro Saeki; Yoshihiko Hosoi; Akira Iritani
    Journal of Mammalian Ova Research 22 (4) 241 - 245 1347-5878 2005 [Refereed]
     
    Recently, with the acetylation of histone and the modification of chromatin structure, the methylation of cytosine residue within CpG dinucleotides in genomic DNA sequence attracts many researchers' attention as one of major epigenetic regulation systems of gene expression. There are several methods (immunofluorescence with 5-methylcytosine specific antibody and methylationsensitive restriction enzyme-PCR method, etc.) to analyze the methylation of cytosine residue. Bisulfitesequencing method, which is one of methods for analyzing the methylation of cytosine residue in genomic DNA sequence, has advantages of high sensitivity and analyzing the methylation of cytosine residue directly. In this note, the detailed procedure of bisulfite-sequencing method for mouse early Preimplantation embryos is described. © 2005, JAPANESE SOCIETY OF OVA RESEARCH. All rights reserved.
  • Mitani T; Nagai T; Suzuki D; Ukida Y; Kato H; Matsumoto K; Saeki K; Hosoi Y; Iritani A
    REPRODUCTION FERTILITY AND DEVELOPMENT C S I R O PUBLISHING 17 (1-2) 208 - 208 1031-3613 2005 [Refereed]
  • Effects of ethanol treatment after intracytoplasmic sperm injection (ICSI) on sperm aster formation and the microtubule organization of bovine oocytes.
    Fujinami N; Hosoi Y; Kato H; Mitani T; Matsumoto K; Saeki K; Hosoi Y; Iritani A
    Reprod. Fertil. Develop. 17 (1) 307  2005/01 [Refereed]
  • Methylation of the 5’-upstream region of the H19 gene in mouse somatic cell, gametes, wild type and androgenetic ES cells
    加藤 博己; 村上秀樹; 川澄みゆり; 國枝孝典; 奥野 学; 岸本 学; 相馬 学; 岩井大悟; 安齋政幸; 三谷 匡; 松本和也; 佐伯和弘; 細井美彦; 入谷 明
    Reproduction, Fertility and Development International Embryo Transfer Society 17 (1,2) 261 - 262 2005/01 
    本研究では、父親由来のゲノム刷り込みの個体発生に対する影響を検討する第一段階として、マウス体細胞、配偶子、2タイプの胚性幹細胞におけるH19遺伝子の5’上流のCpGのメチル化の状態を決定した。ゲノムDNAはBDF1マウスの尾(雌雄体細胞,mSTとfST)、精子(S)、卵子(O)、野生型および雄性発生胚由来胚性幹細胞(wtESとagES)から単離された。CpGのメチル化の状態は、Naバイサルファイト法によって決定された。H19遺伝子の転写開始部位から-4413から-3946塩基の領域には13個のCpGが存在した。個の領域のメチル化CpGの割合は、mSTで88%(160/182)、fSTで79%(27/130)、Sで93%(230/247)、Oで8%(10/130)、wtESで77%(10/13)そしてagESで89%(314/351)であった。CTCF結合部位のコアモチーフ(CCGCGTGGTGGCAG)ではメチル化CpGの割合は、mSTで93%(26/28)、fSTで80%(16/20)、Sで95%(36/38)、Oで0%(0/20)、wtESで50%(1/2)そしてagESで96%(52/54)であった。agESにおけるCpGは精子と同様に高度にメチル化されていた。しかしながら、agESにおいて、既にCpGのメチル化パタ
  • K Saeki; K Matsumoto; M Kinoshita; Suzuki, I; Y Tasaka; K Kano; Y Taguchi; K Mikami; M Hirabayashi; N Kashiwazaki; Y Hosoi; N Murata; A Iritani
    PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA NATL ACAD SCIENCES 101 (17) 6361 - 6366 0027-8424 2004/04 [Refereed]
     
    Linoleic acid (18:2n-6) and a-linolenic acid (18:3n-3) are polyunsaturated fatty acids that are essential for mammalian nutrition, because mammals lack the desaturases required for synthesis of Delta12 (n-6) and n-3 fatty acids. Many plants can synthesize these fatty acids and, therefore, to examine the effects of a plant desaturase in mammals, we generated transgenic pigs that carried the fatty acid desaturation 2 gene for a Delta12 fatty acid desaturase from spinach. Levels of linoleic acid (18:2n-6) in adipocytes that had differentiated in vitro from cells derived from the transgenic pigs were approximate to10 times higher than those from wild-type pigs. In addition, the white adipose tissue of transgenic pigs contained approximate to20% more linoleic acid (18:2n-6) than that of wild-type pigs. These results demonstrate the functional expression of a plant gene for a fatty acid desaturase in mammals, opening up the possibility of modifying the fatty acid composition of products from domestic animals by transgenic technology, using plant genes for fatty acid desaturases.
  • N Fujinami; Y Hosoi; H Kato; K Matsumoto; K Saeki; A Iritani
    JOURNAL OF REPRODUCTION AND DEVELOPMENT SOCIETY REPRODUCTION & DEVELOPMENT-SRD 50 (2) 171 - 178 0916-8818 2004/04 [Refereed]
     
    Developmental potential of bovine embryos that are not artificially activated after intracytoplasmic sperm injection (ICSI) is generally very low. In this study, we investigated effects of artificial activation with ethanol on kinetics of maturation promoting factor (MPF) activity (p34(cdc2) kinase activity) and development of bovine oocytes following ICSI. Treatment of oocytes with ethanol at 4 h after ICSI improved their first cleavage and further preimplantation development (51% vs. 13%, 14% vs. 4%: treatment with vs. without ethanol, respectively). MPF activity of oocytes was lowered until at least 2 h after ICSI. In oocytes without activation after ICSI, MPF activity temporarily elevated at 6 h after ICSI, whereas this phenomena was not observed in the oocytes treated with ethanol. Furthermore, MPF activity was elevated 20 h after ICSI in oocytes activated with ethanol, whereas this elevation of MPF activity was not shown in oocytes without activation. These results indicate that the stimulus of sperm was sufficient to lower MPF activity of oocytes following ICSI, and moreover the activation treatment of bovine oocytes with ethanol after ICSI served to maintain the low levels of MPF activity until the next cell cycle started.
  • 細井 美彦; 松本 和也; 佐伯 和弘; 入谷 明
    Journal of Mammalian Ova Research 20 (1) 34 - 40 2003/01 [Refereed]
     
    ニホンザル過剰排卵システムの確立と顕微授精による体外受精、受精後の胚発生について検討し、報告した。
  • Yoshihiko Hosoi; Ryuzo Torii; Nahoko Fujinami; Kazuya Matsumoto; Kazuhiro Saeki; Akira Iritani
    Journal of Mammalian Ova Research 20 (1) 34 - 40 1347-5878 2003 [Refereed]
     
    Intracytoplasmic Sperm Injection (ICSI) has been widely applied for curing human infertility. In this study the developmental potential of Japanese monkey embryos produced by ICSI is reported in a practically relevant system. Oocytes retrieved by laparoscopy from follicles in ovaries of gonadotrophin-stimulated fertile females were fertilized by ICSI using spermatozoa obtained from a fertile male. An additional chemical stimulus was not necessary to achieve oocyte activation with pronuclear formation after ICSI. Successful fertilization was confirmed by extrusion of the second polar body and the presence of both male and female pronuclei at 18-20 h post-ICSI. Some two-cell stage embryos obtained by ICSI were transferred to synchronous recipients and the others were cultured in CMRL medium for 168 h to assess their developmental competence. Oocytes collected laparoscopically from hyper-stimulated monkey ovaries were fertilized by ICSI and completed preimplantation development in vitro, however no pregnancy was confirmed after embryo transfer. This study demonstrates for the first time that the oocytes of the Japanese monkey are able to support advanced embryonic preimplantation development in vitro. It is suggested that the Japanese monkey is an excellent preclinical model for examining and understanding many aspects of ICSI for endangered primates. © 2003, JAPANESE SOCIETY OF OVA RESEARCH. All rights reserved.
  • Ming Zhang; Kehuan Lu; Kazuya Matsumoto; Yumi Yamamoto; Kazuhiro Saeki; Yoshihiko Hosoi; Akira Iritani; Kazuya Matsumoto; Kazuhiro Saeki; Yoshihiko Hosoi; Akira Iritani
    Journal of Mammalian Ova Research Japanese Society of Mammalian Ova Research 19 (3) 89 - 95 1347-5878 2002 [Refereed]
     
    We identified a novel gene, termed GSE (gonad-specific expression gene). Nucleotide sequence analysis of GSE cDNA revealed that the open reading frame of 745-bp encodes a protein of 247 amino acids with a predicted molecular mass of 27.6 kDa. The deduced amino acid sequence indicated that GSE protein might be a soluble protein in the cytoplasm without a signal peptide. Northern blot analysis showed that this gene was abundantly expressed in mouse testis and slightly expressed in the mouse ovary. RT-PCR analyses indicated that the GSE mRNA in the testis was first detected at Day 14 postpartum, when spermatocytes at mid-pachytene are likely to appear. In situ hybridization confirmed its expression at this stage of spermatogenesis. On the other hand, the GSE mRNA in the ovary was already present at birth, when germ cells are in meiosis. These observations suggest that GSE may be associated with meiosis during gametogenesis. © 2002, JAPANESE SOCIETY OF OVA RESEARCH. All rights reserved.
  • DNA Sequence Retrieval from Ancient Animal Skin Tissue Excavated from Siberian Permafrost
    加藤 博己; 松本 和也; 佐伯 和弘; 細井 美彦; 入谷 明; 米田穣 柴田康行
    Theriogenology 55 (1) 388  2001/01 
    シベリア永久凍土層から出土した古生物皮膚組織からの DNA の抽出と解析を行った。 ミトコンドリア DNA 中に存在するチトクローム b 遺伝子の、 1005 塩基対の塩基配列の決定に成功した。
  • H. Miyamoto; K. Matsumoto; Y. Hosoi; K. Saeki; A. Matsushiro; A. Iritani
    Japanese Journal of Fertility and Sterility 45 347 - 350 0029-0629 2000/01 [Refereed]
     
    We have screened a cDNA library constructed from the rabbit placenta using Mach2 cDNA as a probe to identify genes regulating normal placental development in mammal. We find a cDNA, RPBP1, encoding a novel basic protein that is rich in arginine (13%), lysine (10%), and glutamine (13%). RPBP1 has three transcripts in placenta and the predicted amino acid sequence shows a limited similarity to mouse Mach2. These findings suggest that the RPBP1 protein may have a role in developmental process of placenta correlating the Mach2 function.
  • K Matsumoto; T Nakayama; H Sakai; K Tanemura; H Osuga; E Sato; JE Ikeda
    MOLECULAR REPRODUCTION AND DEVELOPMENT WILEY-LISS 54 (2) 103 - 111 1040-452X 1999/10 [Refereed]
     
    In mammals, the postnatal loss of more than 99% of female germ cells is due mainly to the process of ovarian follicular atresia. Atresia is known to be mediated by apoptotic granulosa cell-death. Here we show the involvement of neuronal apoptosis inhibitory protein (NAIP) in ovarian folliculogenesis in which it prevents granulosa cell-death. NAIP has been isolated in association with a neurodegenerative disorder, spinal muscular atrophy (SMA), in which it has been shown to suppress mammalian cell-death induced by a variety of stimuli (Liston et al., 1996, Nature 379:349-353). In an in situ hybridization analysis with mouse ovaries, active expression of NAIP mRNA was localized in the granulosa cells of developing follicles from the primary stage to the Graafian stages, whereas the NAIP gene was not expressed ou was weakly expressed in follicles that might be undergoing atresia. Gonadotropin, which is known to inhibit apoptosis in ovarian follicles, caused a 2.4-fold increase in NAIP gene expression in the ovary. To study the role of ovarian NAIP, antisense NAIP oligonucleotides were locally delivered into the ovarian bursa. Suppression of ovarian NAIP expression with antisense oligonucleotides evoked a decrease in the number of morphologically normal ovulated oocytes, implying an indirect involvement of NAIP in germ cell development by enhancing the survival of granulosa cells, These findings suggest that gonadotropin-inducible NAIP may indirectly affect oocyte survival as a result of the inhibition of apoptotic granulosa cell-death during folliculogenesis. (C) 1999 Wiley-Liss, inc.
  • Masao Jinno; Yuuko Katsumata; Toshihisa Hoshiai; Yukio Nakamura; Kazuya Matsumoto; Yasunori Yoshimura
    Journal of Clinical Endocrinology and Metabolism Endocrine Society 82 (11) 3603 - 3611 0021-972X 1997 [Refereed]
     
    In a prospective randomized study, we examined whether a novel method of ovarian stimulation, the bromocriptine-rebound method, improves in vitro fertilization (IVF) outcomes compared with the conventional long protocol using GnRH agonist and human menopausal gonadotropin (hMG). Ovulatory women with previous failed IVF-embryo transfer using the long protocol were prospectively assigned to either the bromocriptine-rebound method (group 1, 82 cycles) or the long protocol (group 2, 80 cycles). The bromocriptine- rebound method was the same as the long protocol, except that bromocriptine was administered daily from day 4 of the preceding cycle until 7 days before hMG stimulation. The numbers of follicles, fertilized oocytes, and embryos with superior morphology were higher in group 1 than in group 2. The rates of clinical pregnancy and live birth delivery per cycle were significantly higher in group 1 (38% and 33%, respectively) than in group 2 (21% and 19%, respectively). The mean concentration of serum PRL during hMG administration was significantly higher in group i than group 2. A significant correlation between the number of superior embryos and PRL concentrations was observed in group 1, but not in group 2. Next, we performed a retrospective study to investigate how the bromocriptine-rebound method exerts its beneficial effects. In the initial IVF with the long protocol, the mean concentration of serum PRL during hMG administration and the expression of PRL receptor (PRLr) messenger ribonucleic acid (mRNA) in granulosa cells were significantly higher in nonpregnant patients than in pregnant ones. When IVF was repeated with the bromocriptine-rebound method in the nonpregnant patients, the expression of PRLr mRNA decreased significantly. In conclusion, the bromocriptine-rebound method enhances embryonic development and the rate of live birth delivery in patients with previous failed IVF using the long protocol. We hypothesize that in the nonpregnant patients using the long protocol, the serum PRL concentration and PRLr mRNA expression are increased to compensate for poor postreceptor responsiveness of granulosa cells to PRL during oocyte maturation. The bromocriptine-rebound method may improve oocyte maturation in such patients by restoring postreceptor responsiveness of granulosa cells to PRL during the hypoprolactinemic period and increasing the PRL concentration by a rebound phenomenon after discontinuation of bromocriptine.
  • Kazuya Matsumoto; Seiki Haraguchi; Kazuchika Miyoshi; Akira Awaya; Eimei Sato
    Journal of Reproduction and Development The Japanese Society of Animal Reproduction (JSAR) 43 (2) 137 - 141 1348-4400 1997 [Refereed]
     
    We evaluated the influence of the administration of a synthesized heterocyclic pyrimidine compound, 2-piperadino-6-methyl-5-oxo-5,6-dihydro(7H) pyrrolo-[3,4-d] pyrimidine (MS-818), which has promoting activity of basic fibroblast growth factor(bFGF)-induced angiogenesis, on the number of ovulations in mice (Jcl:ICR strain) superovulated with pregnant mare's serum gonadotropin (PMSG) and human chorionic gonadotropin (hCG). When injected with PMSG and hCG, ovulation failed to be induced in 13.5% of mice, but all of those injected with MS-818 in addition to PMSG and hCG, responded to the stimulation and superovulated. There was also a significant increase in superovulatory response in the mice injected with MS-818 compared with those injected with saline. The mean actual number of ovulations for mice injected with 0 (saline) or 50 μg of MS-818 for 3 days was 13.1 ± 2.1 and 26.9 ± 4.2, respectively, indicating that MS-818 affected the actual number of ovulations. Subcutaneous injection of MS-818 resulted in more actual ovulations than intraperitoneal injection. There was no difference between the control (saline) and MS-818-injected groups in the proportion of ova fertilizing and cleaving to the blastocyst stage in vitro.
  • Matsumoto K
    Tanpakushitsu kakusan koso. Protein, nucleic acid, enzyme 40 (14) 2223 - 2233 0039-9450 1995/10 [Refereed]
  • A TAKAHASHI; Y TAKAHASHI; K MATSUMOTO; K MIYATA
    JOURNAL OF VETERINARY MEDICAL SCIENCE JAPAN SOC VET SCI 57 (3) 553 - 556 0916-7250 1995/06 [Refereed]
     
    To examine effects of IGF-II on establishment of pluripotential diploid cells from rat embryos, we cultured blastocysts in medium containing mouse LIF with or without IGF-II. Combination of mouse LIF (5,000 units/ml) and rat IGF-II (100 ng/ml) promoted growth of inner cell mass (ICM) and was effective for establishment of pluripotential cell lines derived from the ICM. The cell lines indicated colony forms different from the rat ES cell lines. However, they showed morphological alteration to adult-like tissue cells, formed embryoid body in suspension culture, and thus, seemed to retain a pluripotent characteristics. The rat IGF-II is useful for establishing of pluripotential cells efficiently.
  • Kazuya Matsumoto
    Journal of Mammalian Ova Research 12 (2) 65 - 71 1347-5878 1995 [Refereed]
  • K MATSUMOTO; H KAKIDANI; M ANZAI; N NAKAGATA; A TAKAHASHI; Y TAKAHASHI; K MIYATA
    DEVELOPMENTAL GENETICS WILEY-LISS 16 (3) 273 - 277 0192-253X 1995 [Refereed]
     
    We compared the levels of growth hormone (GH) mRNA in the pituitary, plasma GH concentration, and altered phenotype in rats heterozygous and homozygous for an antisense RNA transgene targeted to the rat GH gene, with those in nontransgenic rats. We initially investigated whether the transgene promoter, which is connected to four copies of a thyroid hormone response element (TRE) that increases promoter activity, affected in vivo transgene expression in the pituitary of the transgenic rats. Plasma GH concentration correlated negatively with T-3 injection in surgically thyroidectomized heterozygous transgenic rats. There was a reduction of about similar to 35-40% in GH mRNA levels in the pituitary of homozygous animals compared with those in nontransgenic rats. Plasma GH concentration was significantly similar to 25-32 and similar to 29-41% lower in heterozygous and homozygous transgenic rats, respectively, compared with that in nontransgenic animals. Furthermore, the growth rates in homozygous transgenic rats were reduced by similar to 72-81 and similar to 51-70% compared with those of their heterozygous and nontransgenic littermates, respectively. The results of these studies suggested that the biological effect of GH in vivo is modulated dose-dependently by the antisense RNA transgene. The rat GH gene can therefore be targeted by antisense RNA produced from a transgene, as reflected in the protein and RNA levels. (C) 1995 Wiley-Liss, Inc.
  • K MATSUMOTO; M ANZAI; N NAKAGATA; A TAKAHASHI; Y TAKAHASHI; K MIYATA
    MOLECULAR REPRODUCTION AND DEVELOPMENT WILEY-LISS 39 (2) 136 - 140 1040-452X 1994/10 [Refereed]
     
    We investigated the onset of paternal gene expression in the early mouse embryo. We obtained transgenic mouse embryos by fertilizing BD (C57BL/6N x DBA) F1 hybrid female oocytes in vitro, with sperm from homozygous transgenic males carrying integrated chicken beta-actin promoter-driven firefly luciferase cDNA. We then examined the RNA and protein synthesis of the luciferase gene in embryos from the 1- to 2-cell stage. RNA transcripts of the luciferase gene were first detected in the 1-cell stage embryos as early as 13 hr postinsemination, just prior to elongation. By photon-count imaging, functional luciferase was identified at the 2-cell stage 23 hr postinsemination. These findings indicate that the paternal endogenous gene is already transcribed in the late 1-cell embryos, although paternally derived protein is not synthesized until the 2-cell stage. Therefore, these results suggest that the embryonic gene is activated as early as the late 1-cell stage. (C) 1994 Wiley-Liss, Inc.
  • Anzai M; Nakagata N; Matsumoto K; Ishikawa T; Takahashi Y; Miyata K
    Jikken Dobutsu. 43 (3) 445 - 448 0007-5124 1994/07 [Refereed]
  • Anzai M; Nakagata N; Matsumoto K; Takahashi A; Takahash Y; Miyata K
    Jikken Dobutsu. 43 (2) 247 - 250 0007-5124 1994/04 [Refereed]
  • TAKAHASHI Yumi; TAKAHASHI Akio; MATSUMOTO Kazuya; NAKAGATA Naomi; ANZAI Masayuki; MIYATA Kenji
    Nihon Chikusan Gakkaiho Japanese Society of Animal Science 65 (9) 826 - 833 1346-907X 1994 [Refereed]
     
    With the aim to establish rat embryonic stem (ES) cell lines, we investigated the effects of leukemia inhibiting factor (LIF), buffalo rat liver cell conditioned medium (BRL-CM) and its molecular fractions on the growth of inner cell mass (ICM) from the rat blastocysts. Only the combination of CM/La, which was prepared by ultra filtration with cut-off MW of 5000 as a low molecular fraction from acetic acid-treated BRL-CM, and LIF resulted in dome-like growth of ICM cells at high frequency. By subcultures on feeder layer with the same medium, two cell lines were established. The cell lines showed morphological changes to endoderm-like cells when the CM/La was removed. When both the CM/La and the LIF were removed, the cell lines differentiated into several cell types such as fibroblast-, neuron- and myocardium-like cells, and also formed embryoid body in suspension culture. These results suggest that the cell lines maintain pluripotency and that the CM/La contains both growth-promoting and differentiation-inhibiting activities.
  • Anzai M; Nakagata N; Matsumoto K; Takahasi A; Takahasi Y; Miyata K
    Jikken Dobutsu. Japanese Association for Laboratory Animal Science 42 (3) 467 - 470 0007-5124 1993/07 [Refereed]
     
    Two transgenic mice lines were produced by introducing the rat GH antisense transgene and the chicken β-actin promoter/firefly luciferase hybrid gene in our labolatory. We cryopreserved the transgenic embryos, obtained by fertilization in vitro with the sperm of hemizygous males for the transgene, by the ultrarapid freezing. The survival rates of cryopreserved 2-cell embryos were high (>70%) at thawing in both lines and 53% and 16% of cryopreserved 2-cell embryos, respectively, developed to live young after transferring to oviducts of recipients. Each transgene was detected at about 40% of the live young from two transgenic lines. These results indicated that the cryopreservation of embryos by ultrarapid freezing was valuable for sustaining transgenic mouse lines without genetic contamination.
  • Nakagata N; Matsumoto K; Anzai M; Takahashi A; Takahashi Y; Matsuzaki Y; Miyata K
    Jikken Dobutsu. Japanese Association for Laboratory Animal Science 41 (4) 537 - 540 0007-5124 1992/10 [Refereed]
     
    Spermatozoa of a homozygous tranasgenic mouse, in which the firefly luciferase gene was expressed under the control of β-actin promoter, were frozen at-196°C. One fourth of the frozen sperm was later thawed and used for in vitro fertilization. Thirty-six of 65 oocytes (55.4%) developed to the 2-cell stage. All the 2-cell embryos were transferred to the oviducts of pseudopregnant recipients and 23 young (63.9%, 23/36) were born. All of young analyzed carried the transgene and showed the lucif erase gene expression.
  • A.NAKANISHI; K.MATSUMOTO; K.UTSUMI; A.IRITANI
    Nihon Seishokumeneki Gakkai Zassi Japan Society for Immunology of Reproduction 6 49 - 51 2186-6406 1992
  • 松本 和也; 笠井 健吉; 高橋 明男; 柿谷 均; 宮田 堅司
    日本畜産学会報 日本畜産学会 63 (11) 1163 - 1174 1344-3941 1992 [Refereed]
  • MATSUMOTO Kazuya; MIYAKE Masashi; Utsumi Kyozo; IRITANI Akira
    Journal of Reproduction and Development THE SOCIETY FOR REPRODUCTION AND DEVELOPMENT 33 (1) 1 - 5 0385-9932 1987 [Refereed]
     
    Rat, goat and cattle embryos were bisected by a specially devised metalmicroblade, and subsequent development of the bisected embryos was examined after 0.5-3 h in culture.
    1). High proportions of the embryos from rat, goat and cattle were successfully bisected, but bisection of the goat early blastocysts was comparatively difficult due to that the embryo at this stage still has coarse zona pellucida.
    2). In the goat, it was clarified that the late blastocyst having soft zona pellucida is easily bisected without serious damage of inner cell mass.
    3). The rate of the paired embryos developed subsequently in culture was lower in the rate (29/62, 47%) than in goat (6/6, 100%) or cattle (10/13, 77%).
    These results indicated that bisection of zona intact embryo by metal blade used at the present study is a simple and conventional method, in which both procedures of holding embryo and softening treatment of zona pellucida are not necessary. This method is successfully applied for bisection of goat and cattle late blatocysts, at this stage the zone pellucida has been soft.

Books etc

  • 胚発生プログラムの遺伝的制御
    松本和也; 細井美彦 (Joint work卵子学 (森 崇英 総編集))京都大学学術出版会 2011
  • レポーター遺伝子を用いた初期胚における遺伝子発現の解析(共著)
    生殖工学のための講座 卵子研究法 2001

Conference Activities & Talks

  • 池上春香; 松橋珠子; 永井宏平; 塚口智将; 内堀翔; 樋口智香; 守田昂太郎; 小林直彦; 松本和也; 松本和也
    日本畜産学会大会講演要旨  2016/03
  • 池上春香; 松橋珠子; 永井宏平; 塚口智将; 内堀翔; 樋口智香; 守田昂太郎; 小林直彦; 松本和也; 松本和也
    日本畜産学会大会講演要旨  2015/09
  • 池上春香; 丹羽尚人; 永井宏平; 塚口智将; 樋口智香; 守田昂太郎; 松橋珠子; 小林直彦; 松本和也
    日本畜産学会大会講演要旨  2015/03
  • 池上春香; 松橋珠子; 赤尾大樹; 武本淳史; 永井宏平; 内堀翔; 小林直彦; 松本和也
    日本畜産学会大会講演要旨  2014/03
  • 赤尾大樹; 池上春香; 松橋珠子; 武本淳史; 永井宏平; 樋口智香; 守田昂太郎; 小林直彦; 松本和也
    日本農芸化学会大会講演要旨集(Web)  2014/03
  • 武本淳史; 永井宏平; 池上春香; 樋口智香; 守田昂太郎; 小林栄治; 松本和也
    日本分子生物学会年会プログラム・要旨集(Web)  2012
  • 堀貴裕; 中迫昇; 池上春香; 森本康一; 松本和也; 河本敬子
    電気関係学会関西連合大会講演論文集(CD-ROM)  2011/10
  • 池上春香; 武本淳史; 笹子奈々恵; 小林栄治; 松本和也
    日本畜産学会大会講演要旨  2011/08
  • 武本淳史; 池上春香; 森本康一; 笹子奈々恵; 小林栄治; 松本和也
    日本畜産学会大会講演要旨  2011/08
  • マウス胚性幹細胞の分化過程における細胞核染色体テリトリーと遺伝子座の動態  [Not invited]
    西山 有依; 三谷 匡; 安齋 政幸; 加藤 博己; 松本 和也; 佐伯 和弘; 入谷 明; 細井 美彦; 森木 甲子郎; 藤本 佑希; 田辺 秀之
    第28回日本受精着床学会総会  2010/07  横浜  第28回日本受精着床学会総会
     
    マウス胚性幹細胞の分化過程における細胞核染色体テリトリーと遺伝子座の動態について3D-FISH法を用いて解析した。
  • 池上春香; 松橋珠子; 小林直彦; 大谷健; 森本康一; 松本和也
    日本畜産学会大会講演要旨  2010/03
  • 松本和也; 池上春香; 松橋珠子; 大谷健; 小林直彦; 森本康一
    日本畜産学会大会講演要旨  2010/03
  • 武本淳史; 池上春香; 爲岡奈々恵; 森本康一; 小林栄治; 松本和也
    日本畜産学会大会講演要旨  2010/03
  • 水野陽介; 河本敬子; 池上春香; 森本康一; 松本和也
    情報処理学会全国大会講演論文集  2010/03  東京大学  情報処理学会第72回(平成22)全国大会
  • 池上春香; 園陽平; 永井宏平; 吉廣卓哉; 井上悦子; 小林直彦; 松橋珠子; 大谷健; 中川優; 森本康一; 松本和也
    日本畜産学会大会講演要旨  2009/09
  • 若齢ウシ精巣細胞を用いたヌードマウス皮下への異種異所移植による精子形成誘導の試み  [Not invited]
    森木 甲子郎; 三谷 匡; 安齋 政幸; 加藤 博己; 松本 和也; 佐伯 和弘; 細井 美彦; 入谷 明; 喜多 章太; 藤本 佑希; 谷口 俊仁
    第102回日本繁殖生物学会大会  2009/09  奈良  第102回日本繁殖生物学会大会
     
    若齢ウシ精巣細胞を用いたヌードマウス皮下への異種異所移植による精子形成誘導について試みた。
  • 松本 和也
    The Journal of reproduction and development  2009/08
  • 若齢ウシ精巣細胞を用いたヌードマウス皮下への異種異所移植による精子誘導の試み  [Not invited]
    森木 甲子郎; 三谷 匡; 安齋 政幸; 加藤 博己; 松本 和也; 佐伯 和弘; 細井 美彦; 入谷 明; 喜多 章太; 谷口 俊仁
    第27回日本受精着床学会総会  2009/08  京都  第27回日本受精着床学会総会
     
    若齢ウシ精巣細胞を用いたヌードマウス皮下への異種異所移植による精子誘導について試みた。
  • 天野 朋子; 入谷 明; 松本 和也; 独立行政法人; 産業技術総合研究所; バイオメディカル研究部門; 独立行政法人; 産業技術総合研究所; 生物機能工学研究部門
    Europian Biological Rhythms Society  2009  Strasbourg, France  Europian Biological Rhythms Society
  • 池上春香; 永井宏平; 吉廣卓哉; 園陽平; 小林直彦; 松橋珠子; 大谷健; 中川優; 森本康一; 松本和也
    日本畜産学会大会講演要旨  2008/03
  • SHI Linjing; 井上悦子; 吉廣卓哉; 永井宏平; 池上春香; 松橋珠子; 小林直彦; 森本康一; 松本和也; 中川優
    情報処理学会全国大会講演論文集  2008/03
  • 池上春香; 永井宏平; 吉廣卓哉; 井上悦子; 園陽平; 小林直彦; 松橋珠子; 大谷健; 中川優; 森本康一; 松本和也
    質量分析総合討論会講演要旨集  2008
  • 池上春香; 園陽平; 永井宏平; 吉廣卓哉; 井上悦子; 小林直彦; 松橋珠子; 大谷健; 中川優; 森本康一; 松本和也
    生化学  2008
  • マウス受精卵の第1及び第2分裂機構へのrhophilin-2遺伝子の関与  [Not invited]
    松本 和也; 天野 朋子; 佐伯 和弘; 細井 美彦; 入谷 明; 松岡俊樹; 園陽平; 野老美紀子
    第47回日本哺乳動物卵子学会  2007/05  第47回日本哺乳動物卵子学会
  • Expression of transcription factor Cdx2 and Oct3/4 in mouse somatic cell nuclear transfer embryos.  [Not invited]
    三谷 匡; 西脇恵; 安齋 政幸; 加藤 博己; 松本 和也; 佐伯 和弘; 細井 美彦; 入谷 明
    第40回日本発生生物学会  2007/05  福岡  第40回日本発生生物学会
  • Analysis of global DNA methylation in bovine spermatozoa.  [Not invited]
    加藤 博己; 岸本学; 三谷 匡; 松本 和也; 佐伯 和弘; 細井 美彦; 入谷 明
    The 32nd Annual Conference of the Intenational Embryo Transfer Society  2007  The 32nd Annual Conference of the Intenational Embryo Transfer Society
  • Expression profile and knockdown analysis of a functionally unknown DD2-2 gene in mouse pre-implanataion embryos  [Not invited]
    松本 和也; 申 承旭; 天野 朋子; 佐伯 和弘; 細井 美彦; 入谷 明
    33rd Annual Meeting of Ingternational Embryo Transfer Society  2007/01  33rd Annual Meeting of Ingternational Embryo Transfer Society
  • Identification and characterization of the 5’-flanking region of three mouse maternal genes (Histone H100, Nucleoplasmin2, and Zygote attest1): Transcriptional activity in mouse oocytes.  [Not invited]
    松本 和也; 安齋 政幸; 天野 朋子; 三谷 匡; 加藤 博己; 細井 美彦; 佐伯 和弘; 入谷 明; 常本和伸
    33rd Annual Meeting of Ingternational Embryo Transfer Society  2007/01  33rd Annual Meeting of Ingternational Embryo Transfer Society
  • Effect of aging on amounts of DNA methyltransferase mRNA in mouse spermatozoa.  [Not invited]
    加藤 博己; 甲田俊夫; 岸本学; 三谷 匡; 松本 和也; 佐伯 和弘; 細井 美彦; 入谷 明
    The 33rd Annual Conference of the International Embryo Transfer Society  2007/01  Kyoto, Japan  The 33rd Annual Conference of the International Embryo Transfer Society
  • DNA methylation profiles of upstream elements of Oct3/4 gene in in vitro fertilization (IVF) and somatic cell nuclear-transferred (SCNT) embryos.  [Not invited]
    細井 美彦; 川澄みゆり; 海野祐一; 西脇恵; 松本 和也; 安齋 政幸; 天野 朋子; 三谷 匡; 加藤 博己; 佐伯 和弘; 入谷 明
    The 33rd Annual Conference of the International Embryo Transfer Society  2007/01  Kyoto, Japan  The 33rd Annual Conference of the International Embryo Transfer Society
  • Effects of trichostatin A on development of bovine somatic cell nuclear transfer embryos.  [Not invited]
    佐伯 和弘; 岩本太作; 笠松礼; 立溝篤宏; 阿部悠季; 池田俊太郎; 谷口俊仁; 三谷 匡; 加藤 博己; 松本 和也; 細井 美彦; 入谷 明; 岸上哲士; 若
    The 33rd Annual Conference of the International Embryo Transfer Society  2007/01  Kyoto, Japan  The 33rd Annual Conference of the International Embryo Transfer Society
  • Differentiation of hepatocyte-like cells from mouse embryonic stem cells in a monolayer culture system.  [Not invited]
    三谷 匡; 永井匡; 増谷俊憲; 加藤 博己; 佐伯 和弘; 松本 和也; 細井 美彦; 入谷 明
    The 33rd Annual Conference of the International Embryo Transfer Society  2007/01  Kyoto, Japan  The 33rd Annual Conference of the International Embryo Transfer Society
     
    単層培養によるマウス胚性幹細胞の肝様細胞への分化誘導系について検討した。分化誘導により、初期肝細胞、成熟肝細胞で特異的な遺伝子群の発現が誘導された。また、アルブミン産生、グリコーゲン貯留、アンモニア分解能、尿素合成能などの機能評価を行い、肝機能の一部を獲得していることを示した。
  • トランスジェニックラットより摘出した膝蓋腱の生体力学的特性  [Not invited]
    山本 衛; 松本 和也; 佐伯 和弘; 細井 美彦; 入谷 明
    日本機械学会第19回バイオエンジニアリング講演会  2007/01  仙台  日本機械学会第19回バイオエンジニアリング講演会
     
    遺伝子改変が腱の生体力学的特性に及ぼす影響について検討した。
  • Recovery of viable somatic cells for nuclear transfer from bovine frozen testicles without cryoprotectant.  [Not invited]
    佐伯 和弘; 細井 美彦; 松本 和也; 入谷 明; 岐阜県畜産研究所; 岐阜県畜産研究所; 岐阜県畜産研究所; 岐阜県畜産研究所; 京都大学大学院農学研究科; 近畿大学大学院生物工学専攻; 近畿大学大学院生物工学専攻; 近畿大学大学院生物工学専攻; 近畿大学大学院生物工学専攻
    2007 Annual Conference of Int’l Embryo Transfer Soc  2007/01  京都  2007 Annual Conference of Int’l Embryo Transfer Soc
  • Effects of trichostatin A during in vitro fertilization of bovine oocytes on subsequent development, cell number and allocation of resulting embryos.  [Not invited]
    佐伯 和弘; 池田俊太郎; 立溝篤宏; 岩本太作; 笠松礼; 谷口俊仁; 星野洋一郎; 天野 朋子; 松本 和也; 細井 美彦; 入谷 明
    2007 Annual Conference of Int’l Embryo Transfer Soc  2007/01  Kyoto, Japan  2007 Annual Conference of Int’l Embryo Transfer Soc
  • 池上春香; 永井宏平; 吉廣卓哉; 園陽平; 小林直彦; 松橋珠子; 大谷健; 中川優; 森本康一; 松本和也
    生化学  2007
  • In vitro differentiation of mouse embryonic stem cells into hepatocyte-like cells using monolayer-culture system.  [Not invited]
    三谷 匡; 永井匡; 増谷俊憲; 加藤 博己; 松本 和也; 佐伯 和弘; 細井 美彦; 入谷 明
    The 3rd Annual Conference of Asian Reproductive Biotechnology Society  2006/11  Hanoi, Vietnam  The 3rd Annual Conference of Asian Reproductive Biotechnology Society
     
    単層培養によるマウス胚性幹細胞の肝様細胞への分化誘導系について検討した。分化誘導により、初期肝細胞、成熟肝細胞で特異的な遺伝子群の発現が誘導された。また、アルブミン産生、グリコーゲン貯留、アンモニア分解能、尿素合成能などの機能評価を行い、肝機能の一部を獲得していることを示した。
  • 母性発現遺伝子(Histone H1oo (H1oo), Nucleoplasmin 2 (Npn2), AZygote arrest 1 (Zar1))のプロモーター領域の解析  [Not invited]
    松本 和也; 常本和伸; 天野 朋子; 細井 美彦; 佐伯 和弘; 入谷 明; 安齋 政幸; 三谷 匡; 加藤 博己
    第24回日本受精着床学会総会  2006/09  群馬  第24回日本受精着床学会総会
  • マウス体細胞核移植胚における転写因子Cdx2の発現に関する免疫細胞化学的解析  [Not invited]
    三谷 匡; 西脇恵; 安齋 政幸; 加藤 博己; 松本 和也; 佐伯 和弘; 細井 美彦; 入谷 明
    第24回日本受精着床学会総会  2006/09  第24回日本受精着床学会総会
  • 脂肪酸組成改変クローンウシ作出のための基礎的検討:脂肪分化能によるドナー細胞の検討と遺伝子の導入.  [Not invited]
    佐伯 和弘; 入谷 明; 細井 美彦; 松本 和也; 近畿大学大学院生物工学専攻; 近畿大学大学院生物工学専攻; 近畿大学大学院生物工学専攻; 近畿大学大学院生物工学専攻; 近畿大学大学院生物工学専攻; 近畿大学大学院生物工学専攻
    第99回日本繁殖生物学会大会  2006/09  名古屋  第99回日本繁殖生物学会大会
  • マウス卵巣からのクロマチン解散型卵(NSN)の回収法及び培養法の検討  [Not invited]
    松本 和也; 天野 朋子; 佐伯 和弘; 細井 美彦; 入谷 明; 遠藤 法眞
    第13回日本胚移植研究会大会  2006/08  第13回日本胚移植研究会大会
  • 初期胚におけるNocturninの機能解析  [Not invited]
    松本 和也; 天野 朋子; 佐伯 和弘; 細井 美彦; 入谷 明; 西川 慧
    第13回日本胚移植研究会大会  2006/08  第13回日本胚移植研究会大会
  • Molecular Mechanism for the expression of plasmid-borne reporter gene in mouse pre-implantation embryos  [Not invited]
    松本 和也; 天野 朋子; 佐伯 和弘; 細井 美彦; 入谷 明; 野老美紀子; 山本 由美
    39thAnnual Meeting of the Society for the Study of Reproduction  2006/08  39thAnnual Meeting of the Society for the Study of Reproduction
  • ウシ栄養膜細胞由来核移植胚作製のための体外受精胚のバイオプシー手法の検討  [Not invited]
    佐伯 和弘; 入谷 明; 細井 美彦; 松本 和也; 三谷 匡; 加藤 博己; 近畿大学大学院生物工学専攻; 近畿大学大学院生物工学専攻; 近畿大学大学院生物工学専攻; 近畿大学大学院生物工学専攻; 近畿大学大学院生物工学専攻; 近畿大学大学院生物工学専攻; 近畿大学大学院生物工学専攻
    第13回日本胚移植研究会大会  2006/08  広島市  第13回日本胚移植研究会大会
  • 培養液へのトリコスタチンAの添加がウシ再構築胚の初期発生に及ぼす影響.  [Not invited]
    佐伯 和弘; 入谷 明; 岸上 哲士; 細井 美彦; 松本 和也; 近畿大学大学院生物工学専攻; 近畿大学大学院生物工学専攻; 近畿大学大学院生物工学専攻; 近畿大学大学院生物工学専攻; 近畿大学大学院生物工学専攻; 理化学研究所
    第13回日本胚移植研究会大会  2006/08  広島市  第13回日本胚移植研究会大会
  • Bovine somatic cell nuclear transfer embryos: effects of the gene expression ability and DNA methylation on their development to the blastocyst stage.  [Not invited]
    笠松 礼; 佐伯 和弘; 岩本太作; 谷口俊仁; 三谷 匡; 加藤 博己; 松本 和也; 細井 美彦; 入谷 明; 全農ETセンター; 全農ETセンター; 全農ETセンター
    39th Annual Meeting of the Society for the Study of Reproduction  2006/08  Omaha, NE, USA  39th Annual Meeting of the Society for the Study of Reproduction
  • マウス卵巣におけるプロテオーム解析に関する研究  [Not invited]
    松本 和也; 森本 康一; 佐伯 和弘; 細井 美彦; 入谷 明; 上中崇裕; 永井宏平; 池上春香
    日本ヒトプロテオーム機構(JHUPO)第4回大会・第2回日本臨床プロテオーム研究会(JSCP)連合大会  2006/07  日本ヒトプロテオーム機構(JHUPO)第4回大会・第2回日本臨床プロテオーム研究会(JSCP)連合大会
  • Expression and function of rhopilin-2 gene in the mouse pre-implantaion embryo  [Not invited]
    松本 和也; 天野 朋子; 佐伯 和弘; 細井 美彦; 入谷 明; 松岡 俊樹; 野老美紀子
    20th IUBMB and 11th FAOBMB  2006/06  20th IUBMB and 11th FAOBMB
  • Expression of maternal gene promoter driven expression vectors in mouse oocytes and fertilized eggs.  [Not invited]
    松本 和也; 常本和伸; 早雲真範; 遠藤法真; 安齋 政幸; 天野 朋子; 三谷 匡; 加藤 博己; 細井 美彦; 佐伯 和弘; 入谷 明
    20th IUBMB International Congress of Biochemistry and Molecular Biology and 11th FAOBMB Congress  2006/06  Kyoto, Japan  20th IUBMB International Congress of Biochemistry and Molecular Biology and 11th FAOBMB Congress
  • In vitro culture of CD9-expressing cells enriched by magnetic cell sorting from testes of cryptorchid adult and pup in mice.  [Not invited]
    三谷 匡; 田中裕介; 尾崎嘉昭; 佐伯 和弘; 加藤 博己; 松本 和也; 細井 美彦; 入谷 明
    The 32nd Annual Conference of the International Embryo Society  2006/01  Orlando, USA  The 32nd Annual Conference of the International Embryo Society
  • Relation of spatial gene expression patterns in bovine embryos reconstructed with somatic cells to blastocyst development.  [Not invited]
    佐伯 和弘; 三谷 匡; 加藤 博己; 細井 美彦; 松本 和也; 入谷 明; 玉里友宏; 笠松 礼; 岩本太作; 立溝篤宏; 財; わかやま産業振興財団; 全農ETセンター; 全農ETセンター; 全農ETセンター
    2006 Annual Conference of Int'l Embryo Transfer Soc  2006/01  Orlando, FL, USA  2006 Annual Conference of Int'l Embryo Transfer Soc
  • マウス卵巣におけるプロテオーム解析の確立  [Not invited]
    松本 和也; 天野 朋子; 佐伯 和弘; 細井 美彦; 入谷 明; 森本 康一; 上中崇裕; 永井宏平; 池上春香
    第28回日本分子生物学会  2005/12  第28回日本分子生物学会
  • 飛騨牛白色脂肪組織のプロテオーム解析  [Not invited]
    松本 和也; 入谷 明; 森本 康一; 申承旭; 池上春花; 永井宏平; 上中崇裕; 小林直彦; 大
    第28回日本分子生物学会  2005/12  第28回日本分子生物学会
  • ホウレンソウ由来Δ12脂肪酸不飽和化酵素遺伝子導入マウスにおけるプロテオーム解析  [Not invited]
    松本 和也; 天野 朋子; 佐伯 和弘; 細井 美彦; 入谷 明; 田口 善智; 安齋 政幸; 加藤 博己; 三谷 匡; 新海雄介; 鈴木石根; 村田
    第28回日本分子生物学会  2005/12  第28回日本分子生物学会
  • マウス初期胚におけるrhophilin-2遺伝子の機能解析  [Not invited]
    松本 和也; 天野 朋子; 佐伯 和弘; 細井 美彦; 入谷 明; 松岡俊樹; 早雲真範; 園陽平
    第28回日本分子生物学会  2005/12  第28回日本分子生物学会
  • マウス初期胚における胚性遺伝子発現開始の分子機構  [Not invited]
    松本 和也; 天野 朋子; 佐伯 和弘; 細井 美彦; 入谷 明; 野老美紀子; 山本由美
    第28回日本分子生物学会  2005/12  第28回日本分子生物学会
  • マウス初期胚特異的発現ベクターの開発  [Not invited]
    松本 和也; 天野 朋子; 佐伯 和弘; 細井 美彦; 入谷 明; 安齋 政幸; 加藤 博己; 三谷 匡; 常本和伸
    第28回日本分子生物学会  2005/12  第28回日本分子生物学会
  • マウス体細胞核移植胚におけるoct-4遺伝子転写調節領域のCpGメチル化  [Not invited]
    松本 和也; 天野 朋子; 佐伯 和弘; 細井 美彦; 入谷 明; 安齋 政幸; 加藤 博己; 三谷 匡; 海野佑一; 川澄みゆり
    第28回日本分子生物学会  2005/12  第28回日本分子生物学会
  • G0期および初期G1期の異なる細胞周期のドナー細胞によるウシ再構築胚の遺伝子発現状態と初期発生との関係  [Not invited]
    佐伯 和弘; 入谷 明; 細井 美彦; 松本 和也; 三谷 匡; 近畿大学大学院生物工学専攻; 近畿大学大学院生物工学専攻; 近畿大学大学院生物工学専攻; 近畿大学大学院生物工学専攻; 全農ETセンター; 全農ETセンター; 全農ETセンター
    第28回日本分子生物学会年会  2005/12  福岡市  第28回日本分子生物学会年会
  • 初期G1期細胞によるウシ再構築胚の遺伝子発現状態と初期発生との関係.  [Not invited]
    佐伯 和弘; 入谷 明; 細井 美彦; 松本 和也; 三谷 匡; 加藤 博己; 近畿大学大学院生物工学専攻; 近畿大学大学院生物工学専攻; 近畿大学大学院生物工学専攻; 全農ETセンター; 全農ETセンター; 全農ETセンター
    日本畜産学会第106回大会  2005/12  福岡市  日本畜産学会第106回大会
  • 池上春香; 松本和也; 森本康一; 永井宏平; 上中崇裕; SHIN Sunuku; 小林直彦; 大谷健; 入谷明
    日本分子生物学会年会講演要旨集  2005/11
  • LIMSを用いた生物資源の統合的なプロテオーム解析  [Not invited]
    松本 和也; 森本 康一; 永井宏平; 吉廣卓哉; 剣持聡久; 奥野充利
    日本畜産学会第105回大会  2005/09  日本畜産学会第105回大会
  • ルシフェラーゼ遺伝子導入細胞によるウシ再構築胚の割球での遺伝子発現とその後の初期発生との関係.  [Not invited]
    佐伯 和弘; 岩本太作; 笠松礼; 玉里友宏; 細井 美彦; 松本 和也; 三谷 匡; 加藤 博己; 入谷 明; 谷口俊仁
    日本畜産学会第105回大会  2005/09  札幌  日本畜産学会第105回大会
  • ドナー細胞の細胞周期の違いがウシ再構築胚の遺伝子発現状態と初期発生に及ぼす影響  [Not invited]
    佐伯 和弘; 笠松 礼; 岩本太作; 亀山信二; 玉里友宏; 立溝篤宏; 三谷 匡; 細井 美彦; 松本 和也; 谷口俊仁; 入谷 明; 全農ETセンター; 全農ETセンター; 全農ETセンター
    第98回日本繁殖生物学会大会  2005/09  静岡  第98回日本繁殖生物学会大会
  • 永井宏平; 森本康一; 吉広卓哉; 池上春香; 剣持聡久; 上条憲一; 奥野充利; 中川優; 松本和也
    日本畜産学会大会講演要旨  2005/08
  • 池上春香; 松本和也; 森本康一; 永井宏平; 上中崇裕; 申承旭; 小林直彦; 大谷健; 入谷明
    日本畜産学会大会講演要旨  2005/08
  • マウス初期胚における胚性遺伝子発現の分子機構の検討  [Not invited]
    松本 和也; 佐伯 和弘; 細井 美彦; 入谷 明; 野老美紀子; 山本由美; 申承旭; 松岡俊樹
    第23回日本受精着床学会  2005/08  第23回日本受精着床学会
  • gse遺伝子は減数分裂期の生殖細胞に強く発現する  [Not invited]
    松本 和也; 佐伯 和弘; 細井 美彦; 入谷 明; 水野里志; 松岡 俊樹; 福田愛作; 森本義晴
    第23回日本受精着床学会  2005/08  第23回日本受精着床学会
  • ウシ卵巣の長時間保存が卵子の体外成熟および体外受精後の初期発生におよぼす影響  [Not invited]
    佐伯 和弘; 谷口俊仁; 青葉倫明; 立溝篤宏; 岩本太作; 白水宏一郎; 玉里友宏; 笠松礼; 加藤 博己; 三谷 匡; 松本 和也; 細井 美彦; 入谷 明
    第23回日本受精着床学会  2005/08  大阪  第23回日本受精着床学会
  • ウシ精子頭部DNAのメチル化状態の検討  [Not invited]
    加藤 博己; 岸本学; 三谷 匡; 松本 和也; 佐伯 和弘; 細井 美彦; 入谷 明
    第23回日本受精着床学会  2005/08  大阪  第23回日本受精着床学会
  • マウスAIRE遺伝子の発現と局在に関する組織学的解析  [Not invited]
    三谷 匡; 増田淳大; 松本 和也; 佐伯 和弘; 細井 美彦; 入谷 明
    第23回日本受精着床学会  2005/08  大阪  第23回日本受精着床学会
  • マウス新生仔雄性生殖幹細胞の培養の試み  [Not invited]
    三谷 匡; 田中裕介; 松本 和也; 佐伯 和弘; 細井 美彦; 入谷 明
    第23回日本受精着床学会  2005/08  大阪  第23回日本受精着床学会
  • ウシ体外受精胚盤胞期胚由来栄養膜細胞を用いた核移植  [Not invited]
    佐伯 和弘; 亀山信二; 笠松 礼; 立溝篤宏; 岩本太作; 加藤 博己; 三谷 匡; 松本 和也; 細井 美彦; 入谷 明; 岐阜県畜産研究所; 岐阜県畜産研究所; 岐阜県畜産研究所; 岐阜県畜産研究所
    第12回日本胚移植研究会大会  2005/08  奈良市  第12回日本胚移植研究会大会
  • マウス羊膜上皮細胞における肝特異的遺伝子発現ならびに主要組織適合性抗原の免疫学的応答に関する細胞生物学的解析  [Not invited]
    永井 匡; 三谷 匡; 鈴木 大介; 有喜多 康夫; 松本 和也; 佐伯 和弘; 細井 美彦; 入谷 明
    日本発生生物学会 第38回大会  2005/06  仙台市  日本発生生物学会 第38回大会
     
    羊膜は、胎子を包む膜として発生期にのみ存在する特殊な器官である。羊膜は生理学的に多様な機能を示し、また免疫学的に寛容な性質を有している。本研究では、マウス羊膜上皮細胞を用いて、肝細胞特異的遺伝子発現について検討し、さらにMHC分子の発現誘導に対する応答性について解析した。
  • ルシフェラーゼ遺伝子導入細胞によるウシ再構築胚での遺伝子発現とその後の初期発生との関係  [Not invited]
    佐伯 和弘; 玉里友宏; 笠松礼; 白水宏一郎; 岩本太作; 細井 美彦; 松本 和也; 三谷 匡; 加藤 博己; 谷口俊仁; 入谷 明
    日本畜産学会第104回大会  2005/03  東京  日本畜産学会第104回大会
     
    ウシ再構築胚の発育能と胚内での遺伝子発現との関係を調べるため、ルシフェラーゼ発現ベクターを安定的に導入したウシ繊維芽細胞を用いて再構築胚を作製し、胚内のluc発現とその後の発育能を検討した。
  • アマ由来ω3脂肪酸不飽和化酵素遺伝子のクローニングとその酵素活性の検討.  [Not invited]
    印藤頼子; 佐伯 和弘; 松本 和也; 細井 美彦; 入谷 明; 鈴木石根; 木下幹朗; 三上浩司; 村田紀夫
    日本畜産学会第104回大会  2005/03  東京  日本畜産学会第104回大会
     
    アマ幼若種子からω3脂肪酸不飽和化酵素遺伝子のクローニングし、その酵素活性を酵母を用いて検討した。
  • 植物由来ω3脂肪酸不飽和化酵素遺伝子(FAD3)の動物細胞における酵素活性の解析.  [Not invited]
    白水宏一郎; 佐伯 和弘; 印藤頼子; 立溝篤宏; 細井 美彦; 松本 和也; 入谷 明; 加野浩一郎; 木下幹朗
    日本畜産学会第104回大会  2005/03  東京  日本畜産学会第104回大会
     
    アマ由来ω3脂肪酸不飽和化酵素遺伝子(FAD3)が動物細胞内で機能的に発現するかどうかを検討した。
  • ルシフェラーゼ遺伝子を導入したウシ線維芽細胞を用いたクローン胚における遺伝子発現がその後の初期発生に及ぼす影響  [Not invited]
    玉里友宏; 佐伯 和弘; 笠松礼; 白水宏一朗; 入谷 明; 細井 美彦; 松本 和也; 和歌山県畜産試験場
    日本畜産学会第103回大会  2005/03  府中市  日本畜産学会第103回大会
     
    ルシフェラーゼ遺伝子を導入したウシ繊維芽細胞を用いて再構築胚を作成し、その後のルシフェラーゼ遺伝子発現と胚発生の関係について調べた。その結果、融合直後に発現が見られず、融合60時間後に初めて発現した胚のみが胚盤胞期に発生することがわかった。
  • Characterization of bovine early G1 cells and in vitro development of embryos reconstructed with the cells.  [Not invited]
    笠松 礼; 佐伯 和弘; 玉里友宏; 白水宏一郎; 谷口俊仁; 三谷 匡; 松本 和也; 細井 美彦; 入谷 明; 青柳敬人; 浦川真実; 出田篤司
    2005 Annual Conference of Int’l Embryo Transfer Soc  2005/01  Copenhagen, Denmark  2005 Annual Conference of Int’l Embryo Transfer Soc
     
    初期G1期細胞によるウシ再構築胚がなぜ高い発生率を示すのかを明らかにするため、初期G1期細胞および再構築胚の特徴について免疫組織化学的および分子生物学的手法を持ちいて検討した。
  • Relation of intensity of gene expression in bovine reconstructed embryos to subsequent development.  [Not invited]
    佐伯 和弘; 玉里友宏; 白水宏一郎; 谷口俊仁; 寺井大輔; 松尾純一; 笠松 礼; 松本 和也; 細井 美彦; 入谷 明
    2005 Annual Conference of Int’l Embryo Transfer Soc  2005/01  Copenhagen, Denmark  2005 Annual Conference of Int’l Embryo Transfer Soc
     
    ウシ再構築胚の発育能と胚内での遺伝子発現との関係を調べるため、ルシフェラーゼ発現ベクターを安定的に導入したウシ繊維芽細胞を用いて再構築胚を作製し、胚内のluc発現とその後の発育能を検討した。
  • 天野 朋子; 入谷 明; 松本 和也; Tomomasa Watanabe; Tadashi Mori
    International Embryo Transfer Society  2005/01  コペンハーゲン デンマーク  International Embryo Transfer Society
     
    Increasing of IP3 in the cytoplasm of mammalian oocytes is said to be responsible for [Ca2+]i oscillation observed in the oocytes immediately after sperm penetration, and the [Ca2+]i oscillation is known to be essential for the development of embryos. On the other hand, cumulus cells have been reported to play an important role in cytoplasmic maturation of oocytes and affecting the embryonic development. To obtain more information on the role of cumulus cells in cytoplasmic maturation, the effect of cumulus cells on the rise in [Ca2+]i and on the rate of activation of porcine mature oocytes induced by IP3 injection were investigated.
  • マウス初期胚で発現するrhopilin-2遺伝子と相互作用する因子の探索  [Not invited]
    松本 和也; 天野 朋子; 佐伯 和弘; 細井 美彦; 入谷 明; 安齋 政幸; 加藤 博己; 三谷 匡
    第27回日本分子生物学会  2004/12  神戸  第27回日本分子生物学会
     
    酵母two-hybrid systemにより、rhophilin-2と相互作用する因子を探索し、候補タンパク質については共免疫沈降法により相互作用の有無を確認し、同定した。
  • 植物由来Δ12脂肪酸不飽和化酵素遺伝子導入マウスにおける網羅的遺伝子発現解析  [Not invited]
    松本 和也; 天野 朋子; 佐伯 和弘; 細井 美彦; 入谷 明; 田口 善智; 安齋 政幸; 加藤 博己; 三谷 匡
    第27回日本分子生物学会  2004/12  神戸  第27回日本分子生物学会
     
    脂肪酸不飽和化酵素FAD2を過剰発現するトランスジェニックマウスの表現型に対する遺伝子相互作用の解明を目的として、マイクロアレイを用いた網羅的遺伝子発現解析を行なった。
  • マウス初期胚における時計遺伝子群の発現解析  [Not invited]
    松本 和也; 天野 朋子; 佐伯 和弘; 細井 美彦; 入谷 明; 安齋 政幸; 加藤 博己; 三谷 匡
    第27回日本分子生物学会  2004/12  神戸  第27回日本分子生物学会
     
    マウス初期胚に対する時計遺伝子群の関与について検討することを目的に、本研究では発現プロファイルについて調べた。
  • アマ由来ω3脂肪酸不飽和化酵素遺伝子の酵素活性の検討  [Not invited]
    印藤頼子; 佐伯 和弘; 松本 和也; 細井 美彦; 入谷 明; 基礎生物学研究所; 基礎生物学研究所
    第27回日本分子生物学会年会  2004/12  神戸市  第27回日本分子生物学会年会
     
    アマ幼若種子から既知の配列を元にω3脂肪酸不飽和化酵素遺伝子を単離し酵母へ導入しその酵素活性を検討したところFAD3遺伝子であることが確認できた。さらにこのFAD3遺伝子の哺乳動物細胞での機能を確認するためウシ繊維芽細胞に導入し全脂質の脂肪酸組成を解析したところ、n-3系脂肪酸の上昇が確認され、植物由来のω3脂肪酸不飽和化酵素遺伝子が動物細胞内で機能することが示された。
  • Toxicological studies employing the ES cell lines derived from B57/6 mice  [Not invited]
    笠井重行; 細井 美彦; 川田延幸; 三谷 匡; 佐伯 和弘; 松本 和也; 入谷 明
    The 17th Annual and International Meeting of the Japan Association for Animal Cell Technology 2004  2004/11  Nagoya  The 17th Annual and International Meeting of the Japan Association for Animal Cell Technology 2004
     
    B57/6系マウス由来の胚性幹細胞を用いて、ジメチルニトロソアミンが体外の分化能力に耐える影響を検討した。非常に高い濃度では、神経細胞への分化が抑制された。
  • Production of C57Bl/6 embryonic stem cell derived mice using methods of ICR tetraploid aggregation and 4-cell or blastocyst injection  [Not invited]
    細井 美彦; 川田延幸; 矢持隆之; 安斎政幸; 三谷 匡; 佐伯 和弘; 松本 和也; 入谷 明
    The 17th Annual and International Meeting of the Japan Association for Animal Cell Technology 2004  2004/11  Nagoya  The 17th Annual and International Meeting of the Japan Association for Animal Cell Technology 2004
     
    C57Bl/6胚性幹細胞を樹立し、4倍体胚とのキメラを作製する。この方法により、TGやknock out に頻用されるC57Bl/6胚性幹細胞の寄与率の高い産子を得る方法を確立する。
  • In vitro development of embryos reconstructed with monkey (Macaca Fascicularis) somatic cells and rabbit enucleated oocytes  [Not invited]
    細井 美彦; 矢持隆之; 川田延幸; 松本 和也; 佐伯 和弘; 入谷 明; 太田 聖; 竹; 末森博文; 中辻
    The 17th Annual and International Meeting of the Japan Association for Animal Cell Technology 2004  2004/11  Nagoya  The 17th Annual and International Meeting of the Japan Association for Animal Cell Technology 2004
     
    異種間核移植胚における体外初期発生の発生速度、栄養要求性を検討して報告した。実験結果から、初期発生は細胞質の影響は認められるものの、核の支配が優勢であることが示唆された。
  • アマ由来ω3脂肪酸不飽和化酵素遺伝子のクローニングとその酵素活性の検討  [Not invited]
    印藤頼子; 佐伯 和弘; 入谷 明; 細井 美彦; 松本 和也; 基礎生物学研究所; 基礎生物学研究所; 基礎生物学研究所
    第17回植物脂質シンポジウム  2004/11  つくば市  第17回植物脂質シンポジウム
     
    アマ幼若種子から既知の配列を元にω3脂肪酸不飽和化酵素遺伝子を単離し酵母へ導入しその酵素活性を検討したところFAD3遺伝子であることが確認できた。さらにこのFAD3遺伝子の哺乳動物細胞での機能を確認するためウシ繊維芽細胞に導入し全脂質の脂肪酸組成を解析したところ、n-3系脂肪酸の上昇が確認され、植物由来のω3脂肪酸不飽和化酵素遺伝子が動物細胞内で機能することが示された。
  • カニクイザルの反復過剰排卵法と回収卵子の発生能の検討  [Not invited]
    藤浪菜穂子; 細井 美彦; 加藤博己; 三谷 匡; 松本 和也; 佐伯 和弘; 入谷 明; 竹之下誠
    第22回日本受精着床学会学術講演会・第49回日本不妊学会学術講演会  2004/09  旭川市 9月2-4日  第22回日本受精着床学会学術講演会・第49回日本不妊学会学術講演会
     
    カニクイザルの胚生産供給システムの構築を目指し、過剰排卵法とその後の顕微授精胚の発生について検証した。その結果、カニクイザルの反復過剰排卵処理方法として、rhFSHは有効であると考えられた。
  • ニュージーランド白色種家兎より得られた未成熟卵母細胞の体外培養法  [Not invited]
    是兼真子; 細井 美彦; 佐伯 和弘; 松本 和也; 入谷 明
    第22回日本受精着床学会学術講演会・第49回日本不妊学会総会学術講演会  2004/09  旭川市 9月2-4日  第22回日本受精着床学会学術講演会・第49回日本不妊学会総会学術講演会
     
    卵巣から得られた未成熟卵胞を体外培養し、減数分裂再開能力を獲得させることに成功した。ウサギ前胞状卵胞より得られた卵子卵丘細胞塊は、培養時に立体構造を維持することができるPVP添加培養によって、効率よく発育し、その中に存在するの卵子も高率に減数分裂再開能力を獲得する。
  • Characterization of early G1 cells as nuclear donor for somatic cell cloning in cattle.  [Not invited]
    笠松礼; 佐伯 和弘; 玉里友宏; 白水宏一郎; 寺井大輔; 松尾純一; 入谷 明; 細井 美彦; 松本 和也; 三谷 匡; 和歌山県畜産試験場; 全農ETセンター; 全農ETセンター; 全農ETセンター
    The 3rd Int'l Symposium of the 21st Century COE of Kinki University  2004/09  Wakayama, Japan  The 3rd Int'l Symposium of the 21st Century COE of Kinki University
     
    体細胞クローン胚作成におけるドナー細胞の細胞周期を検討した。
  • Use of bovine preadipocytes as donor cells in somatic cell cloning  [Not invited]
    白水宏一郎; 佐伯 和弘; 玉里友宏; 笠松礼; 寺井大輔; 松尾純一; 岩本太作; 入谷 明; 細井 美彦; 松本 和也; 日本大学生物資源科学部; 和歌山県畜産試験場
    The 3rd Int'l Symposium of the 21st Century COE of Kinki University  2004/09  和歌山市  The 3rd Int'l Symposium of the 21st Century COE of Kinki University
     
    ウシ脂肪細胞から得た前駆脂肪細胞が体細胞クローン胚のドナー細胞として利用できることを示した。
  • Functional Expression of ω3 Fatty Acid Desaturase Gene from Flax Seeds in Transformed Yeasts.  [Not invited]
    印藤頼子; 佐伯 和弘; 入谷 明; 細井 美彦; 松本 和也; 基礎生物学研究所; 基礎生物学研究所; 基礎生物学研究所
    The 3rd Int'l Symposium of the 21st Century COE of Kinki University  2004/09  和歌山市  The 3rd Int'l Symposium of the 21st Century COE of Kinki University
     
    アマ由来omega3脂肪酸不飽和化酵素遺伝子の機能を酵母で検討した。
  • Efficiency of adipocyte conversion of skeletal muscle satellite cells from Japanese Black and Holstein cattle.  [Not invited]
    辻野英和; 佐伯 和弘; 入谷 明; 細井 美彦; 松本 和也
    The 3rd Int'l Symposium of the 21st Century COE of Kinki University  2004/09  和歌山市  The 3rd Int'l Symposium of the 21st Century COE of Kinki University
     
    黒毛和種およびホルスタイン種から採取した筋衛星細胞の脂肪細胞への分化能をin vitroで検討した。
  • 初期G1期のウシ初代繊維芽細胞における遺伝子発現および核の活性状態と核移植後の発生の検討  [Not invited]
    笠松礼; 佐伯 和弘; 玉里友宏; 三谷 匡; 細井 美彦; 松本 和也; 入谷 明; 和歌山県畜産試験場; 全農ETセンター; 全農ETセンター; 全農ETセンター
    第97回日本繁殖生物学会大会  2004/09  東広島市  第97回日本繁殖生物学会大会
     
    ウシ再構築胚作成におけるルシフェラーゼ遺伝子を導入したドナー細胞を用いて、その細胞周期とその後の発生について検討した。その結果、初期G1期に調整した細胞は、遺伝子発現およびKi-67が血清飢餓細胞と同様抑制されているが、細胞周期に不可欠なPCNA活性が高ことが示された。
  • 植物由来脂肪酸不飽和化酵素を発現させたトランスジェニックマウスにおける遺伝子挿入部位の解析  [Not invited]
    松本 和也; 天野 朋子; 田口 善智; 佐伯 和弘; 細井 美彦; 入谷 明; 安齋 政幸; 加藤 博己; 三谷 匡
    第97回日本繁殖生物学会  2004/09  広島  第97回日本繁殖生物学会
     
    トランスジェニックマウスにおける導入遺伝子の染色体上挿入位置を、Inverse PCRを用いて同定した。
  • 生殖腺特異的発現遺伝子gse遺伝子と相互作用をする候補タンパク質の同定  [Not invited]
    松本 和也; 佐伯 和弘; 細井 美彦; 入谷 明; IVF大阪クリニック; IVFなんばクリニック
    第97回日本繁殖生物学会  2004/09  広島  第97回日本繁殖生物学会
     
    マウス卵巣cDNAライブラリーから酵母Two-hybird法をもいた相互作用するタンパク質の探索をした。
  • マウス初期胚における時計遺伝子群の発現解析  [Not invited]
    松本 和也; 天野 朋子; 佐伯 和弘; 細井 美彦; 入谷 明; 安齋 政幸; 加藤 博己; 三谷 匡
    第97回日本繁殖生物学会  2004/09  第97回日本繁殖生物学会
     
    マウス初期胚における時間依存的細胞分裂制御の可能性について検討した。
  • マウス初期胚で発現するrhophilin-2遺伝子と相互作用する因子の探索  [Not invited]
    松本 和也; 天野 朋子; 佐伯 和弘; 細井 美彦; 入谷 明; 安齋 政幸; 加藤 博己; 三谷 匡
    第97回日本繁殖生物学会  2004/09  広島  第97回日本繁殖生物学会
     
    マウス初期胚でその発現が変わる低分子量Gタンパク質伝達に関与するrhophilin-2に着目して、two-hybrid法によって相互作用する因子を探索した。
  • マウス初期胚における胚性遺伝子発現機構へのDNAメチル化およびヒストンアセチル化の関与  [Not invited]
    松本 和也; 天野 朋子; 佐伯 和弘; 細井 美彦; 入谷 明; 安齋 政幸; 三谷 匡; 加藤 博己
    第97回日本繁殖生物学会  2004/09  広島  第97回日本繁殖生物学会
     
    マウス体外受精卵に導入した発現ベクターの発現に対するエピジェネテック修飾の関与について検討した。
  • マウス尾部細胞及びES細胞におけるOct-4遺伝子転写調節領域のCpGメチル化状態の解析  [Not invited]
    松本 和也; 佐伯 和弘; 細井 美彦; 入谷 明
    第11回日本胚移植研究会  2004/09  松山  第11回日本胚移植研究会
     
    oct-4遺伝子の転写調節領域のDNAメチル化状態をマウス尾部組織およびES細胞で、bisulfite-sequencing法によって比較検討した。
  • Δ12脂肪酸不飽和化酵素遺伝子FAD2を発現するトランスジェニックマウス由来細胞における細胞膜流動性の検討  [Not invited]
    松本 和也; 佐伯 和弘; 細井 美彦; 入谷 明
    第11回日本胚移植研究会  2004/09  松山  第11回日本胚移植研究会
     
    トランスジェニックマウス由来細胞の細胞膜流動性をDPHを用いた蛍光偏光光度計による蛍光強度から算出し比較した。
  • マウス初期胚にて発現するnocturnin(DD40)と相互作用するタンパク質の探索  [Not invited]
    松本 和也; 佐伯 和弘; 細井 美彦; 入谷 明
    第11回日本胚移植研究会  2004/09  松山  第11回日本胚移植研究会
     
    酵母Two-hybrid法によって、nocturnin遺伝子と相互作用をする遺伝子の探索を行なった。
  • Two-hybrid systemを用いた生殖腺特異的発現遺伝子gseと相互作用をするタンパク質の同定  [Not invited]
    松本 和也; 佐伯 和弘; 細井 美彦; 入谷 明; IVF大阪クリニック; IVFなんばクリニック
    第22回日本受精着床学会  2004/09  旭川  第22回日本受精着床学会
     
    gse遺伝子がマウス生殖腺で果たす機能を明らかにすることを目的に、相互作用をする遺伝子を検索した。
  • 体細胞クローン胚作製におけるドナー細胞としてのウシ前駆脂肪細胞の利用  [Not invited]
    白水宏一朗; 佐伯 和弘; 玉里友宏; 笠松 礼; 寺井大輔; 松尾純一; 岩本太作; 入谷 明; 細井 美彦; 松本 和也; 日本大学生物資源科学部; 和歌山県畜産試験場
    第11回日本胚移植研究会  2004/08  松山市  第11回日本胚移植研究会
     
    ウシ前駆脂肪細胞が体細胞クローン胚のドナー細胞として利用可能かどうか検討した。
  • 栗原隆; 松本和也; 富永敬一郎; 安斎政幸; 三谷匡; 加藤博己; 佐伯和弘; 細井美彦; 入谷明
    日本畜産学会大会講演要旨  2004/03
  • Effects of cell cycle control methods for donor somatic cells on gene expression and in vitro development of bovine reconsructed embryos.  [Not invited]
    佐伯 和弘; 笠松礼; 玉里友宏; 白水宏一郎; 三谷 匡; 細井 美彦; 松本 和也; 入谷 明; 和歌山県畜産試験場
    The 4th Conference of the Pacific Rim Society for Fertility and Sterility  2004/03  Nago, Okinawa Japan  The 4th Conference of the Pacific Rim Society for Fertility and Sterility
     
    ウシ再構築胚作成におけるルシフェラーゼ遺伝子を導入したドナー細胞を用いて、その細胞周期とその後の発生について検討した。その結果、初期G1期に調整した細胞は、遺伝子発現およびKi-67が血清飢餓細胞と同様抑制されているが、細胞周期に不可欠なPCNA活性が高ことが示された。
  • Kinetics of destabilized luc+ gene expression in mouse preimplantaion embryos  [Not invited]
    松本 和也; 佐伯 和弘; 細井 美彦; 入谷 明; 安齋政幸; 三谷 匡; 加藤 博己
    The 4th Conference of The Pacific Rim Society for Fertility and Sterility  2004/03  Okinawa, Japan  The 4th Conference of The Pacific Rim Society for Fertility and Sterility
     
    マウス初期胚において、半減期の短いdestabilized luciferase遺伝子の発現プロファイルを従来のluciferase遺伝子と比較検討した。
  • Establishment of ES cell lines from diploid androgenetic embryo for producing germline chimera  [Not invited]
    加藤 博己; 村上秀樹; 川澄みゆり; 國枝孝典; 安齋 政幸; 三谷 匡; 松本 和也; 佐伯 和弘; 細井 美彦; 入谷 明
    The 4th Conference of the Pacific Rim Society for Fertility and Sterility  2004/03  沖縄県 名護市  The 4th Conference of the Pacific Rim Society for Fertility and Sterility
     
    雄性のゲノムインプリンティングが個体発生の中でどのような役割をしているのかを検討するために、二倍体雄性発生胚由来の胚性幹細胞を樹立し、そのゲノムにおいてインプリンティングの状態が雄性のインプリンティングが維持されているのか否かを検討した。
  • Effects of gene expression in bovine embryos reconstructed with fibroblasts transfected with luciferase gene on the subsequent development.  [Not invited]
    佐伯 和弘; 玉里友宏; 笠松礼; 白水宏一郎; 松本 和也; 細井 美彦; 入谷 明; 和歌山県畜産試験場
    2004 Annual Conference of Int’l Embryo Transfer Soc  2004/01  Portland, OR, USA  2004 Annual Conference of Int’l Embryo Transfer Soc
     
    ルシフェラーゼ遺伝子を導入したウシ繊維芽細胞を用いて再構築胚を作成し、その後のルシフェラーゼ遺伝子発現と胚発生の関係について調べた。その結果、融合直後に発現が見られず、融合60時間後に初めて発現した胚のみが胚盤胞期に発生することがわかった。
  • ウシ体細胞クローン胚のゲノムDNA におけるCpG メチル  [Not invited]
    松本 和也; 栗原隆; 佐伯 和弘; 細井 美彦; 入谷 明
    第26 回日本分子生物学会年会(神戸)  2003/12  第26 回日本分子生物学会年会(神戸)
     
    ウシ・ゲノム中のCpG 配列のメチル化状態を指標としたウシ体細胞クローン胚の再プログラム化について検討した。
  • Biomechanical properties of cortical bone in rats whose growth hormone gene expression was suppressed by antisense RNA transgene  [Not invited]
    山本 衛; 松本 和也; 佐伯 和弘; 細井 美彦; 入谷 明
    アメリカ機械学会2003 Summer Bioengineering Conference(米国)  2003/06  アメリカ機械学会2003 Summer Bioengineering Conference(米国)
     
    成長ホルモンのアンチセンス遺伝子を導入し、成長ホルモンの分泌を抑制させたトランスジェニックラットの皮質骨の力学的特性を測定した。
  • Relation between the microhardness and physical characteristics of femur in transgenic rats  [Not invited]
    山本 衛; 佐藤隆哉; 松本 和也; 佐伯 和弘; 細井 美彦; 入谷 明
    日本機械学会関西学生会平成14年度卒業研究発表講演会(大阪)  2003/03  日本機械学会関西学生会平成14年度卒業研究発表講演会(大阪)
     
    遺伝子改変が骨の微視的な力学的特性に及ぼす影響を明らかにするとともに、微視的特性と巨視的な骨の物理的性質との関係について検討した。
  • ルシフェラーゼ遺伝子を導入したウシ繊維芽細胞によるクローン胚の遺伝子発現時期の検討  [Not invited]
    佐伯 和弘; 細井 美彦; 松本 和也; 入谷 明
    日本畜産学会第101回大会(つくば)  2003/03  日本畜産学会第101回大会(つくば)
     
    ルシフェラーゼ遺伝子を導入したウシ繊維芽細胞を用いて、クローン胚を作製し、初期発生過程におけるルシフェラーゼ遺伝子発現を検討した。
  • Measurements of the biomechanical properties of cortical bone in transgenic rats  [Not invited]
    山本 衛; 松本 和也; 佐伯 和弘; 細井 美彦; 入谷 明
    日本機械学会第15回バイオエンジニアリング講演会(大阪)  2003/01  日本機械学会第15回バイオエンジニアリング講演会(大阪)
     
    遺伝子改変が骨の生体力学的特性に及ぼす影響について検討した。
  • Molecular cloning and characterization of a novel gene specifically expressed in gonad  [Not invited]
    松本 和也; 佐伯 和弘; 細井 美彦; 入谷 明
    The 29th Annual Conference International Embryo Transfer Society(New Zealand)  2003/01  The 29th Annual Conference International Embryo Transfer Society(New Zealand)
     
    精巣・卵巣で特異的に発現する新規遺伝子GSE(gonad.specific expression gene)を単離とその発現解析について検討した。
  • Atomic force microscopy of bovine acrosome.intact and reacted spermatozoa; their fine structure and numerical analysis  [Not invited]
    佐伯 和弘; 細井 美彦; 松本 和也; 入谷 明
    2003 Annual Conference of Int’l Embryo Transfer Soc(New Zealand)  2003/01  2003 Annual Conference of Int’l Embryo Transfer Soc(New Zealand)
     
    原子間力顕微鏡(AFM)により、先体反応前後のウシ精子頭部の微細構造を観察し、AFMによる3次元解析により先体部分の容積が計測できることを報告した。
  • Development of rabbit preantral follicles isolated from ovaries of sexually matured doe in the scid mice  [Not invited]
    細井 美彦; 松本 和也; 佐伯 和弘; 入谷 明
    The 29th Annual Conference International Embryo Transfer Society(New Zealand)  2003/01  The 29th Annual Conference International Embryo Transfer Society(New Zealand)
     
    ウサギ卵巣から切除された卵胞を免疫不全マウスへ移植し、その発育機序について検討した。卵胞の発育と卵胞のホルモン産成等の機能的な発達と一部ではあるが、卵子の体外成熟が確認された。
  • Two?Hybrid Systemを用いたZAG1と相互作用する遺伝子の探索  [Not invited]
    松本 和也; 佐伯 和弘; 細井 美彦; 入谷 明
    第25回日本分子生物学会年会(横浜)  2002/12  第25回日本分子生物学会年会(横浜)
     
    本実験では、zag1の機能解析を行うと共に、マウス初期胚の発生制御の分子機構を解明することを目的として、ZAG1と相互作用するタンパク質の遺伝子をTwo?Hybrid Systemにより探索した。
  • マウス1細胞期胚における導入外来性遺伝子の転写と翻訳の開始時期の検討  [Not invited]
    松本 和也; 三谷 匡; 安齋 政幸; 佐伯 和弘; 細井 美彦; 入谷 明
    第25回日本分子生物学会年会(横浜)  2002/12  第25回日本分子生物学会年会(横浜)
     
    本実験では、マウス1細胞期胚におけるより詳細な遺伝子発現時期を検討することを目的に、レポーター遺伝子として改良型ホタルルシフェラーゼ遺伝子luc+を前核に顕微注入した胚及びluc+mRNAを細胞質に注入した胚において遺伝子の転写及び翻訳時期を調べた。
  • 子ウシ線維芽細胞への遺伝子導入とクローン胚の作出  [Not invited]
    佐伯 和弘; 細井 美彦; 松本 和也; 入谷 明
    第25回日本分子生物学会年会(横浜)  2002/12  第25回日本分子生物学会年会(横浜)
     
    初代培養のウシ線維芽細胞にマーカー遺伝子を安定的に導入し、その遺伝子導入細胞による再構築胚の作製と導入遺伝子の発現を詳細に検討した。
  • ハムスター顕微授精胚への抗酸化剤の効果  [Not invited]
    細井 美彦; 佐伯 和弘; 松本 和也; 入谷 明
    第123回日本不妊学会関西支部集談会(神戸)  2002/11  第123回日本不妊学会関西支部集談会(神戸)
     
    ハムスターの顕微授精胚に及ぼす抗酸化剤の影響を調べた結果、わずかながらも分割率を改善することが示唆された。
  • Cryotopによるウサギ未受精卵の凍結保存とその後の受精能力の検討  [Not invited]
    細井 美彦; 松本 和也; 佐伯 和弘; 入谷 明; 加藤 博己
    第20回日本受精着床学会学術講演会(岐阜)  2002/10  第20回日本受精着床学会学術講演会(岐阜)
     
    凍結保存の困難な未受精卵を微量メディムとともにガラス化して効率よく保存する方法を検討した。
  • サル未成熟卵子への成熟卵子細胞質注入による体外成熟誘起の検討  [Not invited]
    細井 美彦; 加藤 博己; 佐伯 和弘; 松本 和也; 竹之下誠; 入谷 明
    第20回日本受精着床学会学術講演会(岐阜)  2002/10  第20回日本受精着床学会学術講演会(岐阜)
     
    卵丘細胞を除去したカニクイザル未成熟卵子へ異種(マウス)ならびに同種の成熟卵子細胞質を注入し、体外成熟を誘導した。これらの卵子は、受精・卵割は正常に起こしたが、それ以降の発生は認められなかった。
  • ウサギ2次卵胞のSCIDマウス腎臓被膜下への移植とその発育性に関する研究  [Not invited]
    細井 美彦; 松本 和也; 佐伯 和弘; 入谷 明
    第20回日本受精着床学会学術講演会(岐阜)  2002/10  第20回日本受精着床学会学術講演会(岐阜)
     
    ウサギ卵巣より切出した2次卵胞を免疫不全マウスの腎臓被膜下へ移植し、その発育性を検討した。卵胞は発育したが、卵子の機能は不完全であった。
  • 脂肪組織特異的にホウレンソウ由来ω6脂肪酸不飽和化酵素(FAD2)を発現させたトランスジェニックマウスの解析  [Not invited]
    松本 和也; 田口 善智; 佐伯 和弘; 細井 美彦; 入谷 明
    第95回日本繁殖生物学会(盛岡)  2002/09  第95回日本繁殖生物学会(盛岡)
     
    第93回本大会でオレイン酸をリノール酸に不飽和化するホウレンソウ由来ω6(Δ12)脂肪酸不飽和化酵素fad2 cDNAの上流に脂肪組織特異的プロモーターaP2遺伝子の転写調節領域をつなげた融合遺伝子を導入したトランスジェニックマウス(A系統)の作製について報告した。本実験では、脂肪組織でリノール酸合成能を獲得したトランスジェニックマウスに関して生理学的解析を行った。
  • 植物由来不飽和化酵素遺伝子を構成的に発現させたトランスジェニックマウスにおけるUCP遺伝子発現解析  [Not invited]
    松本 和也; 田口 善智; 佐伯 和弘; 細井 美彦; 入谷 明
    第95回日本繁殖生物学会(盛岡)  2002/09  第95回日本繁殖生物学会(盛岡)
     
    本実験では、エネルギー代謝に着目してTgマウスにおける不飽和化酵素遺伝子の機能的発現による影響についてより詳細な分子的機構を明らかにすること目的に、エネルギー代謝に関係するタンパク質uncoupling protein(脱共役タンパク質、UCP)の発現量をNorthern blot及びWestern blot法により検討した。
  • マウス1細胞期胚における導入外来性遺伝子の転写と翻訳の開始時期の検討  [Not invited]
    松本 和也; 佐伯 和弘; 細井 美彦; 入谷 明
    第95回日本繁殖生物学会(盛岡)  2002/09  第95回日本繁殖生物学会(盛岡)
     
    本実験では、マウス1細胞期胚におけるより詳細な遺伝子発現時期を検討することを目的に、レポーター遺伝子として改良型ホタルルシフェラーゼ遺伝子luc+を前核に顕微注入した胚及びluc+mRNAを細胞質に注入した胚において遺伝子の転写及び翻訳時期を調べた。
  • 脂肪酸不飽和化酵素遺伝子を構成的に発現させたトランスジェニックマウスの生理学的解析  [Not invited]
    松本 和也; 田口 善智; 佐伯 和弘; 細井 美彦; 入谷 明
    第95回日本繁殖生物学会(盛岡)  2002/09  第95回日本繁殖生物学会(盛岡)
     
    本実験では、第91回本大会ですでに報告したfad2遺伝子を構成的に発現させたトランスジェニックマウス(B系統)について、より詳細な表現型を明らかにすることを目的に生理学的検討を加えたので報告した。
  • マウス初期胚におけるDD解析により新規に単離されたzag1遺伝子の抗体を用いた発現解析  [Not invited]
    松本 和也; 佐伯 和弘; 細井 美彦; 入谷 明
    第95回日本繁殖生物学会(盛岡)  2002/09  第95回日本繁殖生物学会(盛岡)
     
    新規に単離されたzag1遺伝子について、推定アミノ酸配列をもとにしたペプチド抗体を作成し、その抗体を用いて発現解析を行った。
  • 外来遺伝子を導入したウシ線維芽細胞によるクローン胚の作出  [Not invited]
    佐伯 和弘; 細井 美彦; 松本 和也; 入谷 明
    第95回日本繁殖生物学会大会(盛岡)  2002/09  第95回日本繁殖生物学会大会(盛岡)
     
    初代培養のウシ線維芽細胞にマーカー遺伝子を安定的に導入し、その遺伝子導入細胞による再構築胚の作製と、胚発生を詳細に検討した。
  • ウシ線維芽細胞胞への遺伝子導入とクローン胚での遺伝子発現  [Not invited]
    佐伯 和弘; 細井 美彦; 松本 和也; 入谷 明
    第9回日本胚移植研究会大会(金沢)  2002/08  第9回日本胚移植研究会大会(金沢)
     
    初代培養のウシ線維芽細胞への遺伝子導入方法の詳細な検討と遺伝子導入細胞を用いた再構築胚の作製を報告した。
  • ルシフェラーゼmRNAを注入したウシ卵子および初期胚における発光検出による翻訳活性の検討  [Not invited]
    佐伯 和弘; 細井 美彦; 松本 和也; 入谷 明
    第9回日本胚移植研究会大会(金沢)  2002/08  第9回日本胚移植研究会大会(金沢)
     
    成熟過程及び成熟卵子また受精直後から前核期における卵子あるいは胚での翻訳活性の有無をルシフェラーゼmRNAを注入して生物発光を調べることで検討した。
  • 顕微注入によるウサギ体細胞核移植の試み  [Not invited]
    細井 美彦; 佐伯 和弘; 松本 和也; 入谷 明
    日本畜産学会第 100 回大会 (武蔵野)  2002/04  日本畜産学会第 100 回大会 (武蔵野)
     
    マウス・クローンの作出に有効であった核の顕微注入法によってウサギ・クローンの作出を試みた。 二例の胎仔を得たが、 産子を得ることはできなかった。
  • REPRESSION OF GENE EXPRESSION AND PRODUCTIVE TRANSLATION AT THE BEGINNING OF MOUSE DEVELOPMENT  [Not invited]
    松本 和也; 佐伯 和弘; 細井 美彦; 入谷 明
    The 27th Annual Conference International Embryo Transfer Society (ブラジル)  2002/01  The 27th Annual Conference International Embryo Transfer Society (ブラジル)
     
    マウス授精直後卵子に存在する転写抑制時期について検討した。
  • Early development and gene expression of reconstructed embryos following fusion of bovine enucleated oocytes with somatic cells from different species  [Not invited]
    佐伯 和弘; 松本 和也; 細井 美彦; 入谷 明
    2002 Annual Conference of Intl Embryo Transfer Soc (ブラジル)  2002/01  2002 Annual Conference of Intl Embryo Transfer Soc (ブラジル)
     
    ルシファラーゼ遺伝子を顕微注入した、 ウシ体細胞クローン胚初期発生における遺伝子発現を、 発光検出により検討した。
  • Application of an internal ribosomal entry sequence (IRES) bicistronic construct to investigate gene function in early mouse development  [Not invited]
    松本 和也; 佐伯 和弘; 細井 美彦; 入谷 明
    第 24 回日本分子生物学会年会 (横浜)  2001/12  第 24 回日本分子生物学会年会 (横浜)
     
    マウス初期胚での遺伝子発現解析の実験系におけるバイシストロン性 IRES 発現ベクターの有用性について事例を示しながら報告した。
  • 異種体細胞による再構築胚の初期発生とレポーター遺伝子の発現開始時期  [Not invited]
    佐伯 和弘; 細井 美彦; 松本 和也; 入谷 明
    第 24 回日本分子生物学会年会 (横浜)  2001/12  第 24 回日本分子生物学会年会 (横浜)
     
    核移植における体細胞核と卵子細胞質の相互作用を調べるためにウシ、 マウスおよびウズラの繊維芽細胞をウシ除核卵子に電気融合した再構築胚を作製し、 それら胚を体外培養して初期発生を調べるとともに、 それぞれの再構築胚にリポーター遺伝子を注入して遺伝子発現時期を調べた。
  • 胚性遺伝子である新規 zga1 遺伝子のタンパク質検出とトランスジェニックマウスの作製  [Not invited]
    松本 和也; 佐伯 和弘; 細井 美彦; 入谷 明
    第 94 回日本繁殖生物学会 (東京)  2001/09  第 94 回日本繁殖生物学会 (東京)
     
    マウス初期胚の発生制御の分子機構を解明することを目的に、 未受精卵と 1 細胞期 G2 期の体外受精卵を供試材料とした蛍光 Differential Displya (Fluoro DD) 法による大量スクリーニングにより数十の遺伝子群の同定を終了し、 その機能解析を進めている。 本実験では、 そのうち新規に単離された遺伝子 zga1 について、 その抗体作成及びトランスジェニックマウス作製を行った。
  • マウス初期胚の遺伝子発現解析におけるバイシストロン性 IRES 発現ベクターの有用性の検討  [Not invited]
    松本 和也; 佐伯 和弘; 細井 美彦; 入谷 明
    第 94 回日本繁殖生物学会 (東京)  2001/09  第 94 回日本繁殖生物学会 (東京)
     
    マウス初期胚の発生制御における分子機構の解析において、 標的遺伝子の gain-of-function あるいは loss-of function によって出現する生理学的影響を探ることは重要な方法である。 本実験では、 初期胚における遺伝子発現解析の実験系を確立することを目的に、 一種類の mRNA から 2 つの ORF を翻訳することができるバイシストロン性 IRES 発現ベクターが初期胚での遺伝子機能解析における有効性について検討した。
  • ウシ培養細胞におけるルシフェラーゼ遺伝子の発光検出による遺伝子発現  [Not invited]
    佐伯 和弘; 細井 美彦; 松本 和也; 入谷 明
    第 51 回関西畜産学会大会 (和歌山)  2001/09  第 51 回関西畜産学会大会 (和歌山)
     
    本報告では、 体細胞クローン胚に広く用いられている繊維芽細胞と卵丘細胞を異なる状態において、 DNA インジェクション法を用いてβ act/luc+の発現検出時期の検討を試みた。
  • ウシ卵子細胞質と異種体細胞による再構築胚の初期発生と遺伝子発現開始時期の検討  [Not invited]
    佐伯 和弘; 細井 美彦; 松本 和也; 入谷 明
    第 94 回日本繁殖生物学会大会 (東京)  2001/09  第 94 回日本繁殖生物学会大会 (東京)
     
    本実験では、 核移植における体細胞核と卵子細胞質の相互作用を調べるためにウシ、 マウスおよびウズラの繊維芽細胞を用いた再構築胚を作製し、 それら胚を体外培養して初期発生を調べるとともに、 それぞれの再構築肺にリポーター遺伝子を注入して遺伝子発現時期を調べた。
  • Ca イオノフォア反復処理による活性化が体細胞核移植胚の初期発生におよぼす影響  [Not invited]
    佐伯 和弘; 細井 美彦; 松本 和也; 入谷 明
    第 8 回日本胚移植研究会大会 (山形)  2001/08  第 8 回日本胚移植研究会大会 (山形)
     
    体細胞核移植では、 卵子細胞質へ体細胞核を移植した後に人為的に活性化することで胚が産子にまで発生することが知られている。 この活性化には Ca イオノフォアがよく利用されているが、 1 回の Ca イオノフォア処理では胚のその後の発生率は大きく向上しない。 そこで、 本実験では、 作製した再構築胚に連続して複数回 Ca イオノフォア処理し、 その後の桑実胚~胚盤胞への発生率を調べるとともに、 胚内の MPF 活性を調べることで、 核移植胚の効率的な活性化処理方法について検討した。
  • ウシ顕微授精胚の発生に及ぼす活性化の影響  [Not invited]
    細井 美彦; 藤浪 菜穂子; 加藤 博己; 松本 和也; 佐伯 和弘; 入谷 明
    第 8 回日本胚移植研究会 (山形)  2001/08  第 8 回日本胚移植研究会 (山形)
     
    ウシ顕微授精を行った後の卵子内 MPF 活性を測定し、 顕微授精後の胚発生と MPF 活性の関係について検討した。
  • マウス初期胚における IRES バイシストロン性発現ベクターの有効性の検討  [Not invited]
    松本 和也; 佐伯 和弘; 細井 美彦; 入谷 明
    第 19 回日本受精着床学会 (横浜)  2001/07  第 19 回日本受精着床学会 (横浜)
     
    1 種類の mRNA から 2 つのオープンリーディングフレームを翻訳することができる (バイシストロン性) 内部リボソーム導入部位 (IRES) を持つ発現ベクターが培養細胞系で使われている。 本実験では、 IRES バイシストロン性発現ベクターが、 初期胚おける有効性について初めて報告した。
  • ウシ卵子細胞質へ異種体細胞を核移植した再構築胚の初期発生  [Not invited]
    佐伯 和弘; 細井 美彦; 松本 和也; 入谷 明
    第 20 回北海道牛受精卵移植研究会および第 12 回西日本胚移植研究会合同研究発表会 (札幌)  2001/07  第 20 回北海道牛受精卵移植研究会および第 12 回西日本胚移植研究会合同研究発表会 (札幌)
     
    核移植後の体細胞核と卵子細胞質の相互作用を調べるために、 ウシ以外にマウス、 さらには哺乳動物とは発生様式の異なる鳥類であるウズラの繊維芽細胞をドナー核として用い、 再構築胚の初期発生について検討した。 融合後の核再形成は融合後 16 時間でいずれの動物の細胞の再構築胚でも核が観察できた。 8 16 細胞期胚の割合は、 マウスではウシより 12 時間速く、 ウズラでは 24 時間遅かった。
  • 抗ブタ・インヒビン抗血清によるハムスター卵胞発育能力の検討  [Not invited]
    細井 美彦; 高橋護; 松本 和也; 佐伯 和弘; 入谷 明
    第 19 回日本受精着床学会 (横浜)  2001/07  第 19 回日本受精着床学会 (横浜)
     
    ブタインヒビンα鎖に対する抗血清を作製し、 ハムスターに投与して胞状卵胞を誘導することを試みた。
  • シベリア永久凍土で回収された古代生物の皮膚からの DNA の抽出と解析■  [Not invited]
    加藤 博己; 松本 和也; 佐伯 和弘; 細井 美彦; 入谷 明; 米田穣 柴田康行
    近畿大学先端技術■合研究所公開シンポジウム (海南)  2001/05  近畿大学先端技術■合研究所公開シンポジウム (海南)
     
    シベリア永久凍土層から出土した古生物皮膚組織から、 ミトコンドリア DNA 中に存在するチトクローム b 遺伝子の、 1127 塩基対の塩基配列の決定に成功し、 比較の結果、 この古生物皮膚組織は現生のスマトラサイに最も近い動物であることが判明した。
  • ANTISENSE INHIBITION OF LUCIFERASE EXPRESSION IN TRANSGENIC MOUSE PREIMPLANTATION EMBRYOS: A COMPARISON OF RNA AND OLIGODEOXYNUCLEOTIDES  [Not invited]
    松本 和也; 佐伯 和弘; 細井 美彦; 入谷 明
    The 27th Annual Conference International Embryo Transfer Society (オマハ)  2001/01  The 27th Annual Conference International Embryo Transfer Society (オマハ)
     
    初期胚における遺伝子発現制御の方法に関する研究について報告した。
  • Onset of gene expression in bovine embryos reconstructed with fibroblasts following reporter gene microinjection  [Not invited]
    佐伯 和弘; 保坂卓; 前田守彦; 南本宏明; 岡本 知可子; 松葉純子; 松本 和也; 細井 美彦; 入谷 明
    2001 Annual Conference on Int'l Embryo Transfer Soc (Omaha, NE, USA)  2001/01  2001 Annual Conference on Int'l Embryo Transfer Soc (Omaha, NE, USA)
     
    体細胞クローン胚の遺伝子発現時期は他受精により得られた胚より早いことを報告した。
  • Hosoi Y; Torii R; Saeki K; Matsumoto K; Iritani A
    Zygote  1998/12

MISC

Industrial Property Rights

Awards & Honors

  • 2004/04 稲盛財団 稲盛財団「研究助成金授与」
     
    受賞者: 松本 和也
  • 2000/07 世界体外受精会議記念賞
     JPN

Research Grants & Projects

  • リキッド・バイオプシー生体予測診断サービス“AIビーフ”の事業化
    国立研究開発法人科学技術振興機構:研究成果展開事業(大学発新産業創出プログラム(START))プロジェクト推進型 起業実証支援
    Date (from‐to) : 2022/10 -2025/03 
    Author : 松本和也
  • 日本学術振興会:科学研究費助成事業 基盤研究(B)
    Date (from‐to) : 2020/04 -2024/03 
    Author : 松本 和也; 根本 充貴; 松橋 珠子
     
    本研究では、我々は肉用牛の肥育期間中に経時的に採取した血清を対象とした質量分析機を用いた定量プロテオミクスSWATH-MS(Sequential window acquisition of all theoretical fragment ion spectra mass spectrometry)解析情報に基づき世界に先駆けて実施している「血清中バイオマーカータンパク質の動態と種類に関する情報から肉用牛の肥育状態を肥育期間中に生体評価するシステム」の開発研究を積み上げている。これは、畜産学と情報科学との異分野融合研究によって、肉用牛の産肉形質を生体評価するシステムの開発と、その結果得られる血清バイオマーカータンパク質の情報に基づく産肉制御の分子機構の統合的理解を目的とするものである。さらに、研究の成果は、これまでその詳細が不明であった肥育期間中の肉用牛における産肉形質の発育に関与する分子機構の解明に資する新しい知見の獲得が期待される。今後、本研究成果の生体評価システムが肉用牛の生産現場に実装された場合、農家が持つ経験値に依存した農業経営から科学的根拠に基づく農業経営への転換の基盤構築に貢献できる。 2年目の成果として、初年度に基盤構築した、日本の地域性の異なる3地域と全国レベルの肉用牛を扱っている研究機関の協力を得て、供試された出荷時の肉用牛(黒毛和種去勢)の枝肉成績の情報と肥育期間中のウシ血清サンプルに基づいた肉用牛の産肉形質の発育状態や肥育状態を生体評価するシステムを用いて、血清バイオマーカータンパク質の情報を基盤とした産肉制御の分子機構の統合的理解を目指して分子間パスウェイネットワーク解析ソフト(IPA)を導入し、産肉形質の形成過程における血清バイオマーカータンパク質の動態から、脂肪交雑の発育シミュレーションモデルを作成した。
  • 肉用牛産肉形質のAI生体評価法の現場実装事業
    日本中央競馬会:畜産振興事業
    Date (from‐to) : 2019/04 -2022/03 
    Author : 松本 和也
  • 肉用牛の産肉形質の生体肥育診断システムの事業化検証
    国立研究開発法人科学技術振興機構:研究成果展開事業(大学発新産業創出プログラム(START))社会還元加速プログラム(SCORE)
    Date (from‐to) : 2020/10 
    Author : 松本和也
  • バイオマーカー解析技術を活用した肉用牛枝肉形質の生体評価手法の確立事業
    日本中央競馬会:畜産振興事業
    Date (from‐to) : 2016/04 -2019/03 
    Author : 松本 和也
  • Japan Society for the Promotion of Science:Grants-in-Aid for Scientific Research Grant-in-Aid for Challenging Exploratory Research
    Date (from‐to) : 2016/04 -2018/03 
    Author : Matsumoto Kazuya
     
    In this study, relations between carcass traits and exosomal miRNA expression profiles in serum were analyzed to identify circulating exosomal miRNA biomarkers for predicting carcass traits and meat quality characteristics of Japanese Black cattle by bioinformatics analysis. Eleven secreted microRNAs were listed as biomarker candidates as a result of microRNA microarray analysis using serum of fattening cattle. Subsequently, reproducibility was confirmed for the two biomarker candidate microRNAs by the association analysis between the amount of blood microRNA quantified by real-time PCR analysis and the carcass traits. Those selected microRNAs were inferred to affect the expression of genes related to differentiation and development of cells constituting muscle tissue by computational searches of existing public databases.
  • エクソソームmiRNAマーカーによる黒毛和種肥育牛の枝肉形質の生体評価は可能か?
    (独)日本学術振興会:科学研究費助成事業 挑戦的萌芽研究
    Date (from‐to) : 2016/04 -2018/03 
    Author : 松本 和也
  • 受精卵のエピジェネティク・リプログラミングへのプロテアソーム分解系の関与機構解明
    (独)日本学術振興会:科学研究費助成事業 基盤研究(B)
    Date (from‐to) : 2013/04 -2017/03 
    Author : 松本 和也
  • Japan Society for the Promotion of Science:Grants-in-Aid for Scientific Research Grant-in-Aid for Challenging Exploratory Research
    Date (from‐to) : 2014/04 -2016/03 
    Author : MATSUMOTO Kazuya; NAGAI Kouhei
     
    In this study, to identify circulating exosomal miRNA biomarkers for predicting carcass traits and meat quality characteristics of Japanese Black cattle by bioinformatics analysis, we first compared exosomal miRNA expression profiles of serum between low and high merit steers in main carcass traits. We then inferred the involvement of the lower- or higher-expressed exosomal miRNAs in metabolic pathways by computational searches of existing public databases (miRDB, and PANTHER). As a result, our study suggests the possibility of at least eight identified exosomal miRNAs as biomarker candidates for predicting carcass traits and meat quality characteristics during fattening of Japanese Black cattle.
  • 黒毛和種肥育牛の肥育期間中に枝肉形質を推定する血中分泌型miRNAマーカーの探索
    (独)日本学術振興会:科学研究費助成事業 挑戦的萌芽研究
    Date (from‐to) : 2014/04 -2016/03 
    Author : 松本 和也
  • 黒毛和種肥育牛の生産性向上とその安定化を実現する新たな肥育診断技術の開発
    日本中央競馬会:畜産振興事業
    Date (from‐to) : 2013/04 -2016/03 
    Author : 松本 和也
  • Japan Society for the Promotion of Science:Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (B)
    Date (from‐to) : 2011/04 -2015/03 
    Author : MINAMI Naojiro; MATSUMOTO Kazuya; TSUKAMOTO Satoshi
     
    Smyd3 is a histone H3 lysine 4 (H3K4) di- and tri-methyltransferase that forms a transcriptional complex with RNA polymerase II and CHD1, which recognizes trimethylated histone H3 lysine 4, is a protein belonging to the family of ATPase-dependent chromatin remodeling factors. In the present study, we investigated the effects of RNA interference (RNAi)-mediated repression on the development of mouse embryos. In Smyd3-, and Chd1-knockdown embryos, the percentage of inner cell mass (ICM)-derived colony formation and trophectoderm (TE)-derived cell attachment was significantly decreased, resulting in a reduction in the number of viable offspring. Furthermore, the expression of Pou5f1, Nanog, and Cdx2, was dramatically decreased in both Smyd3- and Chd1-knockdown embryos. Moreover, in Chd1-knockdown embryos, expression of Hmgpi, which regulate Pou5f1, Nanog, and Cdx2, was also significantly suppressed at zygotic gene activation (ZGA).
  • 哺乳動物初期胚における全能性の分子制御機構の解明
    (独)日本学術振興会:科学研究費助成事業 挑戦的萌芽研究
    Date (from‐to) : 2011/04 -2013/03 
    Author : 松本 和也
  • 黒毛和種の優良経済形質バイオマーカータンパク質の機能解明とモニタリング評価システムの開発
    (独)農業・食品産業技術総合研究機構 生物系特定産業技術研究支援センター(生研センター):イノベーション創出基礎的研究推進事業(発展型)
    Date (from‐to) : 2009/04 -2012/03 
    Author : 松本 和也
  • Japan Society for the Promotion of Science:Grants-in-Aid for Scientific Research Grant-in-Aid for Challenging Exploratory Research
    Date (from‐to) : 2011 -2012 
    Author : MATSUMOTO Kazuya
     
    After fertilization, erasure of the oogenic program and reprogramming by establishing the embryonic programs into totipotent zygote are co-ordinately regulated. However, molecular mechanisms underlying the reprogramming process are not well understood. Understanding of the involvement of proteins in the degradation of maternal mRNA and proteins, and also the onset of ZGA at the maternal-to-zygotic transition helps elucidate molecular mechanisms governing the remodeling of the oocyte into the totipotent zygote and may also have implications for regulation of pluripotency.
  • Japan Society for the Promotion of Science:Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (C)
    Date (from‐to) : 2008 -2010 
    Author : MITANI Tasuku; SAEKI Kazuhiro; MATSUMOTO Kazuya; TAGUCHI Yoshitomo
     
    This study examined the function of ABC transporter, Bcrp1, on the differentiation of mouse ES cells. Bcrp1 defines the side population cell phenotype, which indicates the presumptive functional regulator of stem cells. The results may be summarized as follows : (1) the alternative transcription of Bcrp1 mRNA isoforms and analysis of the cis-element, (2) the upregulation of Oct3/4 gene by Bcrp1 overexpression and its induction of ES cells into endodermal cells, and (3) the possibility of Bcrp1 associating with the products of tumor suppressor gene with Bcrp1 protein.
  • Japan Society for the Promotion of Science:Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (C)
    Date (from‐to) : 2007 -2009 
    Author : HOSOI Yoshihiko; SAEKI Kazuhiro; MATSUMOTO Kazuya
     
    There are a lot of immature follicles in the ovary. However, advances of a follicular development depend on the age, hormonal status and nourishment state of individuals. Therefore a number of full mature follicles is usually limited without superovulation. It is thought that we can produce the next generations efficiently if immature follicles in the ovary can be available. As for the rabbit oocyte in an immature follicle, it is not ready to be fertilized normally. It should be matured in appropriate culture systems to acquire an ability of the normal fertilization and development. In this proposal, we tried to develop a system which make it possible to produce full mature rabbit oocytes in vitro.
  • Japan Society for the Promotion of Science:Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (B)
    Date (from‐to) : 2006 -2009 
    Author : SAEKI Kazuhiro; HOSOI Yoshihiko; MATSUMOTO Kazuya; MITANI Tasuku; TAGUCHI Yoshitomo
     
    Conjugated linoleic acid has been reported to be effective for cancers and obesity. In this study, a gene for an isomerase for conjugated linoleic acid (PAISOM) was introduced into bovine cells. We obtained cells stably transfected with PAISOM gene. We also successfully produced cloned embryos from the PAISOM-cells. However, Gas chromatography of fatty acids in the PAISOM-cells did not indicate the conjugated linoleic acid, but we detected a peak unknown but likely to be conjugated arachidonic acid that were elongated from conjugated linoleic acid.
  • 高品質肉質を持つ和牛のバイオマーカーの探索
    国立研究開発法人 科学技術振興機構:地域イノベーション創出総合支援事業
    Date (from‐to) : 2004 -2009
  • Japan Society for the Promotion of Science:Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (C)
    Date (from‐to) : 2006 -2007 
    Author : MITANI Tasuku; SAEKI Kazuhiro; MATSUMOTO Kazuya; TAGUCHI Yoshitomo
     
    In this research, we aimed to define the role of ABC transporter Bcrpl in the mechanisms of maintenance of pluripotency in mouse embryonic stem cells. Expression profile of Bcrpl protein and mRNA in undifferentiated and differentiated ES cells was examined by flowcytometry and RTP-CR analysis. Moreover, we developed modified dual luciferase reporter assay system to evaluate RNAi efficacy. 1. Correlation of the expression of Bcrpl with Oct3/4 in mouse ES cells. Flowcytometry analysis showed the expression of Bcrpl positively correlated with Oct3/4, a transcription factor regulating undifferentiated status, in mouse ES cells. Moreover, the expression of Bcrpl as well as Oct3/4 in ES cells decreased after in intro differentiation. 2. Expression of Bcrpl mRNA isoforms in ES cells. Mouse Bcrpl mRNA has three isoforms (termed isoform A, B and C) and the differential expression of Bcrpl mRNA isoforms is alternatively regulated in the stem cells and somatic cells. The expression pattern of Bcrpl mRNA isoforms in various tissues in adult mice and undifferentiated ES cells was analyzed by RT-PCR. The transcript of isoform B was strongly expressed in all tissues and ES cells. The isoform A was also expressed in the most tissues and ES cells but isoform C was alternatively transcribed and was quite low in ES cells. After induction of in vitro differentiation, all Bcrpl mRNA isoforms immediately decreased and various differentiation markers appeared thereafter. 3. Development of modified dual luciferase reporter assay system to evaluate RNAi efficacy. In order to establish knockdown ES cell lines against each isoforms of Bcrpl mRNA for functional analysis, we developed modified dual luciferase reporter assay system to evaluate RNAi efficacy. Reporter-based luciferase assay system represented positive correlation with Western blot analysis in the reduction of target protein by shRNA expression vector in mouse ES cells.
  • Japan Society for the Promotion of Science:Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (B)
    Date (from‐to) : 2003 -2005 
    Author : SAEKI Kazuhiro; HOSOI Yoshihiko; MATSUMOTO Kazuya; MITANI Tasuku
     
    The purpose of this study is to produce cattle containing healthy polyunsaturated fatty acids that are not biosynthesized by mammals. To achieve this purpose, we developed a method to produce transgenic cattle carrying a gene for a fatty acid desaturase taken from plants that are able to newly biosynthesize the polyunsaturated fatty acids. We obtained an FAD3 cDNA which encoded an ω3 fatty acid desaturase from flax seeds that contained the highest rate of α-linolenic acid (18:3n-3) contents in land plants. New synthesis of 18:3n-3 was observed when the cDNA was introduced into yeast and examined their fatty acid composition. Furthermore, a transfection method of a foreign gene stably into mammalian primary cultured cell was established. Using this method, pβ-act/FAD3/IRES/EGFP(neor) was transfected into bovine fibroblast cells and mouse 3T3-L1 cells, and then the functional expression was examined by determination of composition rates of 18:3n-3. The FAD3 gene derived from flax seeds expressed functionally in mammalian cultured cells, because the ratio of the 18:3n-3 increased in the transfected cells with the FAD3 gene compared with that in wild-type cells. However, the ratio in the transfected cells were low and only 0.4% (wild type : 0.2%). Therefore, the DNA sequence of the FAD3 that was a plant type was changed to the mammalian-type DNA sequence according to the mammalian codon usage. Then we obtained a new FAD3 gene optimized in mammalian cells (FAD3opt), and expression vector (pCAG/FAD3opt/IRES/EGFP(neor)) was produced. Strong expression of EGFP was observed in the mouse 3T3-L1 cells transfected with pCAG/FAD3opt/IRES/EGFP(neor). In addition, we have obtained a bovine primary fibroblast cell lines carrying pCAG/FAD3opt/IRES/EGFP(neor). We now examine the fatty acid composition in the transfected cells. We also examined the relation of gene expression in bovine cloned embryos to their further development, to establish an effective method for production of cloned calves with the transgenic cells as donor cells. We examined gene expression in embryos reconstructed with bovine fibroblasts carrying a luciferase expression vector (pβ-act/luc+/IRES/EGFP(neor)). As a result, when intensity of LUC+ expression was examined in transgenic cloned embryos 60 hours post fusion (hpf), strongly expressed-embryos developed at a high rate. Moreover, when The embryos were then classified as being Luc-positive, mosaic, or Luc-negative depending on whether all, some or no blastomeres were luminescent, respectively, the developmental rates to the blastocyst stage of positive embryos were higher than those of mosaic embryos. Any negative embryos did not developed to blastocysts. Then, we examined DNA methylation levels in the embryos by immunostaining with a 5-methyl cytosine antibody. The methylation level of the Luc-positive embryos was lower than that of the Luc-negative embryos. These results indicated that the developmental capacity of NT embryos could be correlated positively to the gene expression ability and negatively to the DNA methylation level at 4- to 8-cell stage.
  • Japan Society for the Promotion of Science:Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (C)
    Date (from‐to) : 2003 -2005 
    Author : MITANI Tasuku; SAEKI Kazuhiro; HOSOI Yoshihiko; MATSUMOTO Kazuya
     
    In order to develop a novel method to produce genetically-modified food animals, we aimed to separate somatic stem cells in various tissues applicable for donor cells for producing cloned mice. We also examined an establishment of ES cells from nuclear transferred embryos and their ability to differentiate. Moreover, we examined manipulating an expression of certain target gene. 1.Multipotential amniotic epithelial cells (AECs) were cultured and characterized their properties. AECs showed moderate expression of MHC class I and II and the expression of those molecules by IFN-? was only slightly induced. It was notably that AECs expressed some major hepatic genes. Spermatogonial stem cells (SSC) were enriched from testes using MACS for CD9 or ?6-integrin. SSCs could be cultured in vitro for more than 2 months with expression of CD9, ?6-integrin and Oct4 genes. 2.As aire-deficient mice showed insufficiency of gametogenesis, the expression of aire was examined in gonads and ES cells. AIRE proteins were partially distributed in immature thymus, ovary and ES cells and localized in the nuclei forming nuclear dots. 3.AECs from EGFP-transgenic mice were examined for nuclear transfer. Using the reconstituted embryos which developed to the blastocyst stage, ES cells were established. ES cells were also established from parthenotes and were confirmed their ability to differentiate in vitro and in vivo. 4.Embryonic fibroblasts, cumulus cells, ES cells and AECs were used for producing cloned mouse. Cloned mouse derived from cumulus cells was produced. Factors affecting developmental ability of reconstituted embryos and conditions for improvement of their development were investigated. 5.RNAi-expression vector for SOD1, mutant protein of which induces ALS, was introduced into ES cells and using its stable transfectant, knockdown mice for SOD1 were produced. By crossing this SOD1-siRNA transgenic mouse with human ALS model mouse, siRNA prevented the development ALS.
  • Japan Society for the Promotion of Science:Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (C)
    Date (from‐to) : 2000 -2002 
    Author : SAEKI Kazuhiro; MATSUMOTO Kazuya; HOSOI Yoshihiko
     
    The following research was done for the production of transforming growth factor 8 gene (TGF8) knock-out cattle using a chimeric DNA/RNA oligo-nucleotide. 1) To investigate effects of cell cycle of the recipient cells, bovine fibroblasts and cumulus cells, on the gene expression, p-β-act/luc^+/IRES/EGFP/neo^r was injected into the cell nuclei. The luminescence was highly detected when cells were in a confluent state, but not after serum starvation. To examine the efficiency of gene transformation of the cells, gene introduction methods, DNA-microinjection, etectroporation and a transfection reagent were used for introduction of the p-β-act/luc^+/IRES/EGFP/neo^r into the bovine somatic cells. The gene was transfected most effectively by use of the transfection reagent by detecting EGFP fluorescence. Furthermore, in the transfected cells, the fluorescence was further detected after selection by G418, and in nuclear transplanted-embryos with the transfected. These indicated that the gene may be integrared in genome of the cells. From the data, the use of the transfection reagent was the most effective to introduce the exogenous gene into bovine somatic cells. 2) A point mutation of cyh2 encoding L29 which is a ribosomal protein in yeast induce cychloheximide tolerance. We examined whether the similar mutation of the homologous gene encoding L27, a mammalian ribosomal protein induced the cychloheximide tolerance during cell culture. We designed the chimeric DNA/RNA oligo-nucleotide which induced the mutation of L27 gene, and introduced the oligo-nucleotide into bovine and mouse fibroblasts by the transfection reagent The cells were cultured under cychloheximide for 10 days. All of the not-transfected and vecter-transfected cells were degenerated 10 days after culture with cychloxeximide, but bovine and mouse cells transfected with the oligo-nucleotide were survived even with cychloheximide for 10 days. The results showed the transfection of the oligo-nucleotide might induce the point-mutation of the L27 gene of the cells and consequently the cells acquired the cychloheximide tolerance. 3) We designed the the chimeric DNA/RNA oligo-nucleotide which induced point-mutation of bovine GDF8 gene. The use of the oligo-nucleotide may provide the alternative method to produce the KO animals.
  • Japan Society for the Promotion of Science:Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (C)
    Date (from‐to) : 2000 -2002 
    Author : HOSOI Yoshihiko; MATSUMOTO Kazuya; SAEKI Kazuhiro; IRITANI Akira
     
    During fertilization, sperm entry triggers a series of intracellular calcium oscillations critical to oocyte activation. This activation is prerequisite to the resumption and completion of meiosis, subsequent pronuclear formation and DNA synthesis. Intracytoplasmic sperm injection (ICSI) has been applied to human infertility treatment. This technique could be also useful for determining the roles of various sperm components in fertilization. Although ICSI is usually a highly successful treatment for severe male factor infertility, some cases repeatedly experience low ICSI fertilization rates (<50%). Possible causes include failure of oocyte activation resulting from sperm and/or oocyte defects. Previous studies have shown that oocytes injected with round spermatids (Hosoi et al., 2000) can be activated by the calcium ionophore A23187. Various artificial treatments mimic sperm-triggered events and induce parthenogenetic development in MII oocytes. It suggests that additional stimulation helps development of fertilized eggs under insufficient condition. In considering the application of intracytoplasmic sperm injection (ICSI) for endangered species and infertility patients, in many cases only not adequate gametes are expected to be used for reproduction. In conclusion, our study showed that oocytes activation factors was very important for artificial fertilization.
  • 日本学術振興会:科学研究費助成事業 萌芽的研究
    Date (from‐to) : 1998 -1999 
    Author : 佐伯 和弘; 松本 和也; 細井 美彦
     
    ウシ形質転換胚を受卵雌に移植する前に選別するため、ルシフェラーゼ遺伝子をウシ胚に注入し初期発生過程での胚の発光検出の有効性を検討している。すでに、遺伝子注入胚における発現は授精後60時間以降開始することを報告している。しかしながら、120時間以降その発現率が低下することから、初期の発現はほとんどが一過性と思われた。安定的な発現を検出するため、今回、まず、遺伝子注入胚の胚盤胞期での発現検出を試みた。二ワトリβアクチン/ホタルルシフェラーゼ(β-act/lUc)融合遺伝子を、体外成熟・受精で作製したウシ胚前核に注入した。これらのの胚を7日間培養して胚盤胞期まで発生させ、イメ一ジングフォトンカウンターを用いて胚の発光を観察した。1,402個の胚にβ-act/lUc遺伝子を注入したところ、63個(4.5%)が胚盤胞に発生したが、発育胚において発光は観察されなかった。今回遺伝子発現が全く検出されなかったことは、用いたβ-act/lUc遺伝子のウシ胚内でめ発現効率が低いことも考えられた。そこで、SV40由来プロモータ/エンハンサーを有する改良型ルシフェラーゼ(SV/luc+)遺伝子およびニワトリβアクチンプロモータを有する改良型ルシフェラーゼ(β-act/lUc+)遺伝子をウシ胚に注入し、その発現効率を検討し、効率よく発現するレポータ遺伝子を検索した。遺伝子は、プラスミドを用いた。SV/luc+およびβ-act/lUc+注入胚は、授精後60時間以降で発光が検出され、発光した胚の割合は、それぞれ、28.5および20.0%といずれも高率だった。また、発光強度は、以前用いたβ-act/lUcより強かった。以上より、より効率に生物発光を検出するには、改良型ルシフェラーゼ遺伝子が適していると思われた。


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