KINDAI UNIVERSITY


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MATSUMOTO Kazuya

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FacultyDepartment of Genetic Engineering / Graduate School of Biology-Oriented Science and Technology
PositionProfessor
Degree
Commentator Guidehttps://www.kindai.ac.jp/meikan/488-matsumoto-kazuya.html
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Last Updated :2020/09/14

Education and Career

Education

  •  - 1989 03 , Kyoto University
  •  - 1989 , Kyoto University, Graduate School, Division of Agriculture
  •  - 1986 03 , Kyoto University
  •  - 1983 03 , Utsunomiya University, Faculty of Agriculture
  •  - 1983 , Utsunomiya University, Faculty of Agriculture

Research Activities

Research Areas

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

Research Interests

  • Molecular Developmental Biology, Reproductive Biology

Published Papers

  • Signs of biological activities of 28,000-year-old mammoth nuclei in mouse oocytes visualized by live-cell imaging., 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, Scientific reports, 9(1), 4050, Mar. 2019 , Refereed
  • Evaluation of antioxidant status and oxidative stress markers in follicular fluid for human in vitro fertilization outcome., Nishihara T, Matsumoto K, Hosoi Y, Morimoto Y, Reproductive medicine and biology, Reproductive medicine and biology, 17(4), 481 - 486, Oct. 2018 , Refereed
  • 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, 関西畜産学会報, (175), 1 - 9, Mar. 2018 , Refereed
  • Ubiquitin-proteasome system modulates zygotic genome activation in early mouse embryos and influences full-term development., 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, The Journal of reproduction and development, 64(1), 65 - 74, Feb. 2018 , Refereed
  • Large-scale proteomic analysis of Japanese Black cattle: Part III. Identification of protein biomarker candidates for assessment of carcass traits and meat quality characteristics, 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, 日本畜産学会報, 86(2), 141 - 152, May 25 2015 , Refereed
  • Large-scale proteomic analysis of Japanese Black cattle: Part II. Assessment of gender and genetic background effects, 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, 日本畜産学会報, 83(3), 281 - 290, Aug. 25 2012 , Refereed
  • Large-scale proteomic analysis of Japanese Black cattle: Part II. Assessment of gender and genetic background effects, 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, 日本畜産学会報, 83(3), 281 - 290, Aug. 25 2012
  • Involvement of the circadian clock into the cyclicity of the mammalian females, 天野 朋子, 入谷 明, 松本 和也, Japanese Society for Chronobiology, Japanese Society for Chronobiology, 16, 31 - 41, 2010
  • Functional expression of a humanized gene for an omega-3 fatty acid desaturase from scarlet flax in transfected bovine adipocytes and bovine embryos cloned from the cells., 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, Biochimica et biophysica acta, 1791(3), 183 - 190, Mar. 2009 , Refereed
  • Developing an integrated database system for the large-scale proteomic analysis of Japanese Black cattle, 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, 日本畜産学会報, 79(4), 467 - 481, Nov. 25 2008 , Refereed
  • Developing an integrated database system for the large-scale proteomic analysis of Japanese Black cattle, 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, 日本畜産学会報, 79(4), 467 - 481, Nov. 25 2008
  • Investigation of in vitro fertilization and early embryonic vitrification in JAX │MICE C57BL/6J, 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, 実験動物技術, 40(2), 87 - 90, Dec. 2005 , Refereed
  • Fine surface structure of bovine acrosome-intact and reacted spermatozoa observed by atomic force microscopy., Saeki K, Sumitomo N, Nagata Y, Kato N, Hosoi Y, Matsumoto K, Iritani A, J Reprod Dev, J Reprod Dev, 51(2), 293 - 298, Apr. 2005 , Refereed
  • Activation with ethanol improves embryo development of ICSI-derived oocytes by regulation of kinetics of MPF activity., Fujinami N, Hosoi Y, Kato H, Matsumoto K, Saeki K, Iritani A, J Reprod Dev., J Reprod Dev., 50(2), 171 - 178, Apr. 2004 , Refereed
  • Fertilization by Intracytplasmic Sperm Injection and Subsequent Embryo Development In Vitro Blastocysts In Japanese Monkey(Macaca fuscata), 細井 美彦, 松本 和也, 佐伯 和弘, 入谷 明, Journal of Mammalian Ova Research, Journal of Mammalian Ova Research, 20(1), 34 - 40, Jan. 2003 , Refereed
    Summary: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。
  • Sequence of a cDNA encoding a novel basic protein expressed in rabbit placenta, H. Miyamoto, K. Matsumoto, Y. Hosoi, K. Saeki, A. Matsushiro, A. Iritani, Japanese Journal of Fertility and Sterility, Japanese Journal of Fertility and Sterility, 45, 347 - 350, Jan. 01 2000 , Refereed
    Summary: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.
  • [Transgenic rat]., Matsumoto K, Tanpakushitsu kakusan koso. Protein, nucleic acid, enzyme, Tanpakushitsu kakusan koso. Protein, nucleic acid, enzyme, 40(14), 2223 - 2233, Oct. 1995 , Refereed
  • [Production of transgenic mice from in vitro fertilized eggs cryopreserved by ultrarapid freezing]., Anzai M, Nakagata N, Matsumoto K, Ishikawa T, Takahashi Y, Miyata K, Jikken Dobutsu., Jikken Dobutsu., 43(3), 445 - 448, Jul. 1994 , Refereed
  • [Cryopreservation of in vitro fertilized embryos from transgenic rat by ultrarapid freezing]., Anzai M, Nakagata N, Matsumoto K, Takahashi A, Takahash Y, Miyata K, Jikken Dobutsu., Jikken Dobutsu., 43(2), 247 - 250, Apr. 1994 , Refereed
  • [Cryopreservation of transgenic mouse embryos by ultrarapid freezing]., Anzai M, Nakagata N, Matsumoto K, Takahasi A, Takahasi Y, Miyata K, Jikken Dobutsu., Jikken Dobutsu., 42(3), 467 - 470, Jul. 1993 , Refereed
  • [Cryopreservation of spermatozoa of a transgenic mouse]., Nakagata N, Matsumoto K, Anzai M, Takahashi A, Takahashi Y, Matsuzaki Y, Miyata K, Jikken Dobutsu., Jikken Dobutsu., 41(4), 537 - 540, Oct. 1992 , Refereed
  • Peroxiredoxin as a functional endogenous antioxidant enzyme in pronuclei of mouse zygotes, 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, Journal of Reproduction and Development, 64(2), 161 - 171, 2018 , Refereed
    Summary: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.
  • Histone H3 Methylated at Arginine 17 Is Essential for Reprogramming the Paternal Genome in Zygotes, 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 REPORTS, 20(12), 2756 - 2765, Sep. 2017 , Refereed
    Summary: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.
  • Reprogramming towards totipotency is greatly facilitated by synergistic effects of small molecules, 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, BIOLOGY OPEN, 6(4), 415 - 424, Apr. 2017 , Refereed
    Summary: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.
  • The Clock mutation reduces reproductive performance of mice by affecting the implantation capacity: Maternal Clock mutation is not the only factor affecting implantation, Tomoko Amano, Masayuki Anzai, Kazuya Matsumoto, THERIOGENOLOGY, THERIOGENOLOGY, 86(7), 1670 - 1684, Oct. 2016 , Refereed
    Summary: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.
  • Production of somatic cell nuclear transfer embryos using in vitro-grown and in vitro-matured oocytes in rabbits, Hironobu Sugimoto, Yuta Kida, Noriyoshi Oh, Kensaku Kitada, Kazuya Matsumoto, Kazuhiro Saeki, Takeshi Taniguchi, Yoshihiko Hosoi, ZYGOTE, ZYGOTE, 23(4), 494 - 500, Aug. 2015 , Refereed
    Summary: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.
  • Possible Role of ZPAC, Zygote-specific Proteasome Assembly Chaperone, During Spermatogenesis in the Mouse, 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, JOURNAL OF REPRODUCTION AND DEVELOPMENT, 60(3), 179 - 186, Jun. 2014 , Refereed
    Summary: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.
  • Oral melatonin supplementation improves oocyte and embryo quality in women undergoing in vitro fertilization-embryo transfer, Takuji Nishihara, Shu Hashimoto, Keijiro Ito, Yoshiharu Nakaoka, Kazuya Matsumoto, Yoshihiko Hosoi, Yoshiharu Morimoto, GYNECOLOGICAL ENDOCRINOLOGY, GYNECOLOGICAL ENDOCRINOLOGY, 30(5), 359 - 362, May 2014 , Refereed
    Summary: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.
  • Development of interspecies cloned embryos reconstructed with rabbit (Oryctolagus cuniculus) oocytes and cynomolgus monkey (Macaca fascicularis) fibroblast cell nuclei, 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, ZYGOTE, 21(4), 358 - 366, Nov. 2013 , Refereed
    Summary: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.
  • Nicotinamide: a Class III HDACi Delays In Vitro Aging of Mouse Oocytes, Ah Reum Lee, Satoshi Kishigami, Tomoko Amano, Kazuya Matsumoto, Teruhiko Wakayama, Yoshihiko Hosoi, JOURNAL OF REPRODUCTION AND DEVELOPMENT, JOURNAL OF REPRODUCTION AND DEVELOPMENT, 59(3), 238 - 244, Jun. 2013 , Refereed
    Summary: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.
  • Functional Analysis of Nocturnin, a Circadian Deadenylase, at Maternal-to-zygotic Transition in Mice, 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, JOURNAL OF REPRODUCTION AND DEVELOPMENT, 59(3), 258 - 265, Jun. 2013 , Refereed
    Summary: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.
  • GSE Is a Maternal Factor Involved in Active DNA Demethylation in Zygotes, 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, PLOS ONE, 8(4), e60205, Apr. 2013 , Refereed
    Summary: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.
  • Dynamics and regulation of lysine-acetylation during one-cell stage mouse embryos, Keigo Matsubara, Ah Reum Lee, Satoshi Kishigami, Akio Ito, Kazuya Matsumoto, Hongfang Chi, Norikazu Nishino, Minoru Yoshida, Yoshihiko Hosoi, BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS, BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS, 434(1), 1 - 7, Apr. 2013 , Refereed
    Summary: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.
  • Mouse zygote-specific proteasome assembly chaperone important for maternal-to-zygotic transition, 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, BIOLOGY OPEN, 2(2), 170 - 182, Feb. 2013 , Refereed
    Summary: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.
  • The ubiquitin-proteasome system in the maternal-to-zygotic transition, Seung-Wook Shin, Shigeo Murata, Kazuya Matsumoto, Journal of Mammalian Ova Research, Journal of Mammalian Ova Research, 30(3), 79 - 85, 2013
    Summary: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.
  • Donor Cells at the G1 Phase Enhance Homogeneous Gene Expression Among Blastomeres in Bovine Somatic Cell Nuclear Transfer Embryos, Daisaku Iwamoto, Aya Kasamatsu, Atsushi Ideta, Manami Urakawa, Kazuya Matsumoto, Yoshihiko Hosoi, Akira Iritani, Yoshito Aoyagi, Kazuhiro Saeki, CELLULAR REPROGRAMMING, CELLULAR REPROGRAMMING, 14(1), 20 - 28, Feb. 2012 , Refereed
    Summary: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.
  • Predicting three-way interactions of proteins from expression profiles based on correlation coefficient, Etsuko Inoue, Sho Murakami, Takatoshi Fujiki, Takuya Yoshihiro, Atsushi Takemoto, Haruka Ikegami, Kazuya Matsumoto, Masaru Nakagawa, IPSJ Transactions on Bioinformatics, IPSJ Transactions on Bioinformatics, 5, 34 - 43, 2012 , Refereed
    Summary: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.
  • Identification and characterization of an oocyte factor required for development of porcine nuclear transfer embryos, 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, PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 108(17), 7040 - 7045, Apr. 2011 , Refereed
    Summary: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.
  • Deposition of Acetylated Histones by RNAP II Promoter Clearance May Occur at Onset of Zygotic Gene Activation in Preimplantation Mouse Embryos, 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, JOURNAL OF REPRODUCTION AND DEVELOPMENT, 56(6), 607 - 615, Dec. 2010 , Refereed
    Summary: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
  • Inhibition of the Ubiquitin-proteasome System Leads to Delay of the Onset of ZGA Gene Expression, 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, JOURNAL OF REPRODUCTION AND DEVELOPMENT, 56(6), 655 - 663, Dec. 2010 , Refereed
    Summary: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
  • Expression analysis of circadian genes in oocytes and preimplantation embryos of cattle and rabbits, 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, ANIMAL REPRODUCTION SCIENCE, 121(3-4), 225 - 235, Sep. 2010 , Refereed
    Summary: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.
  • Potential Existence of Stem Cells With Multiple Differentiation Abilities to Three Different Germ Lineages in Mouse Neurospheres, Toshiyuki Takehara, Takeshi Teramura, Yuta Onodera, Satoshi Kishigami, Kazuya Matsumoto, Kazuhiro Saeki, Kanji Fukuda, Yoshihiko Hosoi, STEM CELLS AND DEVELOPMENT, STEM CELLS AND DEVELOPMENT, 18(10), 1433 - 1440, Dec. 2009 , Refereed
    Summary: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.
  • Enhancement of histone acetylation by trichostatin A during in vitro fertilization of bovine oocytes affects cell number of the inner cell mass of the resulting blastocysts, Shuntaro Ikeda, Atsuhiro Tatemizo, Daisaku Iwamoto, Shunji Taniguchi, Yoichiro Hoshino, Tomoko Amano, Kazuya Matsumoto, Yoshihiko Hosoi, Akira Iritani, Kazuhiro Saeki, ZYGOTE, ZYGOTE, 17(3), 209 - 215, Aug. 2009 , Refereed
    Summary: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.
  • Proteomic Analysis of the Mouse Ovary in Response to Two Gonadotropins, Follicle-Stimulating Hormone and Luteinizing Hormone, 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, JOURNAL OF REPRODUCTION AND DEVELOPMENT, 55(3), 316 - 326, Jun. 2009 , Refereed
    Summary: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.
  • Mouse Androgenetic Embryonic Stem Cells Differentiated to Multiple Cell Lineages in Three Embryonic Germ Layers In Vitro, 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, JOURNAL OF REPRODUCTION AND DEVELOPMENT, 55(3), 283 - 292, Jun. 2009 , Refereed
    Summary: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.
  • Abnormal DNA Methylation of the Oct-4 Enhancer Region in Cloned Mouse Embryos, 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, MOLECULAR REPRODUCTION AND DEVELOPMENT, 76(4), 342 - 350, Apr. 2009 , Refereed
    Summary: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.
  • Expression and Functional Analyses of Circadian Genes in Mouse Oocytes and Preimplantation Embryos: Cry1 Is Involved in the Meiotic Process Independently of Circadian Clock Regulation, 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, BIOLOGY OF REPRODUCTION, 80(3), 473 - 483, Mar. 2009 , Refereed
    Summary: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.
  • Isolation and Culture of Rabbit Primordial Germ Cells, 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, JOURNAL OF REPRODUCTION AND DEVELOPMENT, 54(5), 352 - 357, Oct. 2008 , Refereed
    Summary: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.
  • Cis-acting elements (E-box and NBE) in the promoter region of three maternal genes (Histone H1oo, Nucleoplasmin 2, and Zygote arrest 1) are required for oocyte-specific gene expression in the mouse, 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, MOLECULAR REPRODUCTION AND DEVELOPMENT, 75(7), 1104 - 1108, Jul. 2008 , Refereed
    Summary: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.
  • Identification of ZAG1, a novel protein expressed in mouse prei rn plantation, and its putative roles in zygotic genome activation, 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, JOURNAL OF REPRODUCTION AND DEVELOPMENT, 54(3), 192 - 197, Jun. 2008 , Refereed
    Summary: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.
  • H1foo is indispensable for meiotic maturation of the mouse oocyte, 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, JOURNAL OF REPRODUCTION AND DEVELOPMENT, 53(4), 895 - 902, Aug. 2007 , Refereed
    Summary: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.
  • Mitochondrial dysfunction of in vitro grown rabbit oocytes results in preimplantation embryo arrest after activation, Hiroyuki Kanaya, Shu Hashimoto, Takeshi Teramura, Yoshiharu Morimoto, Kazuya Matsumoto, Kazuhiro Saeki, Akira Iritani, Yoshihiko Hosoi, JOURNAL OF REPRODUCTION AND DEVELOPMENT, JOURNAL OF REPRODUCTION AND DEVELOPMENT, 53(3), 631 - 637, Jun. 2007 , Refereed
    Summary: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.
  • Abnormal distribution of chromosomes in the first division of nuclear transferred mouse embryos, Miyuri Kawasumi, Masayuki Anzai, Toshiyuki Takehara, Tasuku Mitani, Hiromi Kato, Kazuhiro Saeki, Akira Iritani, Kazuya Matsumoto, Yoshihiko Hosoi, JOURNAL OF REPRODUCTION AND DEVELOPMENT, JOURNAL OF REPRODUCTION AND DEVELOPMENT, 53(3), 615 - 622, Jun. 2007 , Refereed
    Summary: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.
  • Primate embryonic stem cells proceed to early gametogenesis in vitro, 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, Cloning and Stem Cells, 9(2), 144 - 156, 2007 , Refereed
    Summary: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.
  • A mouse and embryonic stem cell derived from a single embryo, 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, Cloning and Stem Cells, 9(4), 485 - 494, 2007 , Refereed
    Summary: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.
  • Application of laser-assisted zona drilling to in vitro fertilization of cryopreserved mouse oocytes with spermatozoa from a subfertile transgenic mouse, 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, JOURNAL OF REPRODUCTION AND DEVELOPMENT, 52(5), 601 - 606, Oct. 2006 , Refereed
    Summary: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.
  • Expression and subcellular localization of GSE protein in germ cells and preimplantation embryos, Satoshi Mizuno, Youhei Sono, Toshiki Matsuoka, Kazuya Matsumoto, Kazuhiro Saeki, Yoshihiko Hosoi, Aisaku Fukuda, Yoshiharu Morimoto, Akira Iritani, JOURNAL OF REPRODUCTION AND DEVELOPMENT, JOURNAL OF REPRODUCTION AND DEVELOPMENT, 52(3), 429 - 438, Jun. 2006 , Refereed
    Summary: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.
  • Dynamic rearrangement of telomeres during spermatogenesis in mice, K Tanemura, A Ogura, C Cheong, H Gotoh, K Matsumoto, E Sato, Y Hayashi, HW Lee, T Kondo, DEVELOPMENTAL BIOLOGY, DEVELOPMENTAL BIOLOGY, 281(2), 196 - 207, May 2005 , Refereed
    Summary: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.
  • Functional expression of a Delta 12 fatty acid desaturase gene from spinach in transgenic pigs, 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, PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 101(17), 6361 - 6366, Apr. 2004 , Refereed
    Summary: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.
  • Fertilization by Intracytoplasmic Sperm Injection and Subsequent Embryo Development in Vitro to Blastocysts in Japanese Monkey (Macaca Fuscata), Yoshihiko Hosoi, Ryuzo Torii, Nahoko Fujinami, Kazuya Matsumoto, Kazuhiro Saeki, Akira Iritani, Journal of Mammalian Ova Research, Journal of Mammalian Ova Research, 20(1), 34 - 40, 2003 , Refereed
    Summary: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.
  • Molecular cloning and characterization of a novel gene specifically expressed in gonad, 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, Journal of Mammalian Ova Research, 19(3), 89 - 95, 2002 , Refereed
    Summary: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.
  • Neuronal apoptosis inhibitory protein (NAIP) may enhance the survival of granulosa cells thus indirectly affecting oocyte survival, K Matsumoto, T Nakayama, H Sakai, K Tanemura, H Osuga, E Sato, JE Ikeda, MOLECULAR REPRODUCTION AND DEVELOPMENT, MOLECULAR REPRODUCTION AND DEVELOPMENT, 54(2), 103 - 111, Oct. 1999 , Refereed
    Summary: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.
  • A therapeutic role of prolactin supplementation in ovarian stimulation for in vitro fertilization: The bromocriptine-rebound method, Masao Jinno, Yuuko Katsumata, Toshihisa Hoshiai, Yukio Nakamura, Kazuya Matsumoto, Yasunori Yoshimura, Journal of Clinical Endocrinology and Metabolism, Journal of Clinical Endocrinology and Metabolism, 82(11), 3603 - 3611, 1997 , Refereed
    Summary: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.
  • Enhancement of number of ovulations by administration of a pyrimidine compound with potentiating activity of angiogenesis in mice superovulated by PMSG and hCG, Kazuya Matsumoto, Seiki Haraguchi, Kazuchika Miyoshi, Akira Awaya, Eimei Sato, Journal of Reproduction and Development, Journal of Reproduction and Development, 43(2), 137 - 141, 1997 , Refereed
    Summary: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.
  • Activation of the Early Zygotic Genome in the Mouse Embryo, Kazuya Matsumoto, Journal of Mammalian Ova Research, Journal of Mammalian Ova Research, 12(2), 65 - 71, 1995 , Refereed
  • Synergistic effects of insulin-like growth factor II (IGF-II) with leukemia inhibiting factor (LIF) on establishment of rat pluripotential cell lines., Takahashi A, Takahashi Y, Matsumoto K, Miyata K, J Vet Med Sci., J Vet Med Sci., 57(3), 553 - 556, 1995 , Refereed
  • EVALUATION OF AN ANTISENSE RNA TRANSGENE FOR INHIBITING GROWTH-HORMONE GENE-EXPRESSION IN TRANSGENIC RATS, K MATSUMOTO, H KAKIDANI, M ANZAI, N NAKAGATA, A TAKAHASHI, Y TAKAHASHI, K MIYATA, DEVELOPMENTAL GENETICS, DEVELOPMENTAL GENETICS, 16(3), 273 - 277, 1995 , Refereed
    Summary: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.
  • ONSET OF PATERNAL GENE ACTIVATION IN EARLY MOUSE EMBRYOS FERTILIZED WITH TRANSGENIC MOUSE SPERM, K MATSUMOTO, M ANZAI, N NAKAGATA, A TAKAHASHI, Y TAKAHASHI, K MIYATA, MOLECULAR REPRODUCTION AND DEVELOPMENT, MOLECULAR REPRODUCTION AND DEVELOPMENT, 39(2), 136 - 140, Oct. 1994 , Refereed
    Summary: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.

Conference Activities & Talks

  • 黒毛和種去勢牛の枝肉形質を生体評価するバイオマーカー候補タンパク質の解析, 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 27
  • 黒毛和種牛の枝肉形質を予測するバイオマーカー候補タンパク質の肥育期間中の動態, 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 11
  • 黒毛和種牛の脂肪交雑度を予測する血清マイクロRNAの探索, 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 28
  • 黒毛和種牛の脂肪交雑度を予測するタンパク質バイオマーカーの探索, 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 27
  • 血清のプロテオーム解析による黒毛和種牛の脂肪交雑度を診断するバイオマーカーの探索, 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 05
  • ウシ僧帽筋のプロテオーム解析による筋内脂肪量に関わる新規タンパク質バイオマーカー候補の同定, 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
  • 黒毛和種牛の形質と影響タンパク質に対するSVMの検討, 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 17
  • ウシ筋肉内脂肪組織タンパク質の網羅的解析, 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 26
  • 黒毛和種とリムジン種の交雑家系(F2)のウシ僧帽筋のプロテオーム解析による経済形質バイオマーカーの探索, 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 26
  • ウシ血清中に含まれるタンパク質の網羅的解析方法の確立, 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 28
  • ウシの枝肉形質に関与するバイオマーカー候補タンパク質のパスウェイ解析, 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 28
  • ウシ筋肉中タンパク質の網羅的プロテオーム解析, 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 28
  • 黒毛和種牛の形質と影響タンパク質に対する多変量解析の検討, 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 08
  • ウシの経済形質に関与する新規バイオマーカーAnnexin A5アイソフォームの同定, 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 28
  • Expression and functional analysis of circadian genes in mouse ovaries, Europian Biological Rhythms Society,   2009 , Europian Biological Rhythms Society
  • 黒毛和牛の肉質に関連するバイオマーカーの探索, 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 27
  • ブランド牛(飛騨牛)の繁殖農家のための種雄牛選択支援システム, 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 13
  • ウシの枝肉形質を規定するバイオマーカーの探索―質量分析を用いたタンパク質アイソフォームの解析―, 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
  • 黒毛和牛の枝肉形質を診断するバイオマーカー候補タンパク質の探索:白色脂肪組織の大規模プロテオーム解析, 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
  • Expression of transcription factor Cdx2 and Oct3/4 in mouse somatic cell nuclear transfer embryos.,   2007 05
  • Analysis of global DNA methylation in bovine spermatozoa., 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, 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., 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., The 33rd Annual Conference of the International Embryo Transfer Society,   2007 01 , 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., The 33rd Annual Conference of the International Embryo Transfer Society,   2007 01 , The 33rd Annual Conference of the International Embryo Transfer Society
  • Effects of trichostatin A on development of bovine somatic cell nuclear transfer embryos., The 33rd Annual Conference of the International Embryo Transfer Society,   2007 01 , 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., The 33rd Annual Conference of the International Embryo Transfer Society,   2007 01 , The 33rd Annual Conference of the International Embryo Transfer Society
  • Recovery of viable somatic cells for nuclear transfer from bovine frozen testicles without cryoprotectant., 2007 Annual Conference of Int’l Embryo Transfer Soc,   2007 01 , 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., 2007 Annual Conference of Int’l Embryo Transfer Soc,   2007 01 , 2007 Annual Conference of Int’l Embryo Transfer Soc
  • 優良肉質を規定するバイオマーカー候補の探索:飛騨牛をモデルとして, 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., The 3rd Annual Conference of Asian Reproductive Biotechnology Society,   2006 11 , The 3rd Annual Conference of Asian Reproductive Biotechnology Society
  • Molecular Mechanism for the expression of plasmid-borne reporter gene in mouse pre-implantation embryos, 39thAnnual Meeting of the Society for the Study of Reproduction,   2006 08 , 39thAnnual Meeting of the Society for the Study of Reproduction
  • Bovine somatic cell nuclear transfer embryos: effects of the gene expression ability and DNA methylation on their development to the blastocyst stage., 39th Annual Meeting of the Society for the Study of Reproduction,   2006 08 , 39th Annual Meeting of the Society for the Study of Reproduction
  • Expression and function of rhopilin-2 gene in the mouse pre-implantaion embryo, 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., 20th IUBMB International Congress of Biochemistry and Molecular Biology and 11th FAOBMB Congress,   2006 06 , 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., The 32nd Annual Conference of the International Embryo Society,   2006 01 , 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., 2006 Annual Conference of Int'l Embryo Transfer Soc,   2006 01 , 2006 Annual Conference of Int'l Embryo Transfer Soc
  • 飛騨牛白色脂肪組織のプロテオーム解析, 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 25
  • LIMSを用いた生物資源の統合的なプロテオーム解析, 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 25
  • 飛騨牛白色脂肪組織のプロテオーム解析, 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 25
  • Characterization of bovine early G1 cells and in vitro development of embryos reconstructed with the cells., 2005 Annual Conference of Int’l Embryo Transfer Soc,   2005 01 , 2005 Annual Conference of Int’l Embryo Transfer Soc
  • Relation of intensity of gene expression in bovine reconstructed embryos to subsequent development., 2005 Annual Conference of Int’l Embryo Transfer Soc,   2005 01 , 2005 Annual Conference of Int’l Embryo Transfer Soc
  • The effct of cumulus cells during maturation on the rise in the concentration of intracellular Ca2+([Ca2+]i) of porcine oocytes induced by inositol 1,4,5-trisphosphate, International Embryo Transfer Society,   2005 01 , International Embryo Transfer Society
    Summary: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.
  • Toxicological studies employing the ES cell lines derived from B57/6 mice, The 17th Annual and International Meeting of the Japan Association for Animal Cell Technology 2004,   2004 11 , The 17th Annual and International Meeting of the Japan Association for Animal Cell Technology 2004
  • Production of C57Bl/6 embryonic stem cell derived mice using methods of ICR tetraploid aggregation and 4-cell or blastocyst injection, The 17th Annual and International Meeting of the Japan Association for Animal Cell Technology 2004,   2004 11 , The 17th Annual and International Meeting of the Japan Association for Animal Cell Technology 2004
  • In vitro development of embryos reconstructed with monkey (Macaca Fascicularis) somatic cells and rabbit enucleated oocytes, The 17th Annual and International Meeting of the Japan Association for Animal Cell Technology 2004,   2004 11 , The 17th Annual and International Meeting of the Japan Association for Animal Cell Technology 2004
  • Characterization of early G1 cells as nuclear donor for somatic cell cloning in cattle., The 3rd Int'l Symposium of the 21st Century COE of Kinki University,   2004 09 , The 3rd Int'l Symposium of the 21st Century COE of Kinki University
  • Use of bovine preadipocytes as donor cells in somatic cell cloning, The 3rd Int'l Symposium of the 21st Century COE of Kinki University,   2004 09 , The 3rd Int'l Symposium of the 21st Century COE of Kinki University
  • Functional Expression of ω3 Fatty Acid Desaturase Gene from Flax Seeds in Transformed Yeasts., The 3rd Int'l Symposium of the 21st Century COE of Kinki University,   2004 09 , The 3rd Int'l Symposium of the 21st Century COE of Kinki University
  • Efficiency of adipocyte conversion of skeletal muscle satellite cells from Japanese Black and Holstein cattle., The 3rd Int'l Symposium of the 21st Century COE of Kinki University,   2004 09 , The 3rd Int'l Symposium of the 21st Century COE of Kinki University
  • ウシ体細胞クローンはいにおける機能的遺伝子の転写調節領域のCpGメチル化, 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 20
  • Effects of cell cycle control methods for donor somatic cells on gene expression and in vitro development of bovine reconsructed embryos., The 4th Conference of the Pacific Rim Society for Fertility and Sterility,   2004 03 , The 4th Conference of the Pacific Rim Society for Fertility and Sterility
  • Kinetics of destabilized luc+ gene expression in mouse preimplantaion embryos, The 4th Conference of The Pacific Rim Society for Fertility and Sterility,   2004 03 , The 4th Conference of The Pacific Rim Society for Fertility and Sterility
  • Establishment of ES cell lines from diploid androgenetic embryo for producing germline chimera, The 4th Conference of the Pacific Rim Society for Fertility and Sterility,   2004 03 , The 4th Conference of the Pacific Rim Society for Fertility and Sterility
  • Effects of gene expression in bovine embryos reconstructed with fibroblasts transfected with luciferase gene on the subsequent development., 2004 Annual Conference of Int’l Embryo Transfer Soc,   2004 01 , 2004 Annual Conference of Int’l Embryo Transfer Soc
  • Biomechanical properties of cortical bone in rats whose growth hormone gene expression was suppressed by antisense RNA transgene,   2003 06
  • Relation between the microhardness and physical characteristics of femur in transgenic rats,   2003 03
  • Measurements of the biomechanical properties of cortical bone in transgenic rats,   2003 01
  • Molecular cloning and characterization of a novel gene specifically expressed in gonad, The 29th Annual Conference International Embryo Transfer Society(New Zealand),   2003 01 , The 29th Annual Conference International Embryo Transfer Society(New Zealand)
  • Atomic force microscopy of bovine acrosome.intact and reacted spermatozoa; their fine structure and numerical analysis, 2003 Annual Conference of Int’l Embryo Transfer Soc(New Zealand),   2003 01 , 2003 Annual Conference of Int’l Embryo Transfer Soc(New Zealand)
  • Development of rabbit preantral follicles isolated from ovaries of sexually matured doe in the scid mice, The 29th Annual Conference International Embryo Transfer Society(New Zealand),   2003 01 , The 29th Annual Conference International Embryo Transfer Society(New Zealand)
  • REPRESSION OF GENE EXPRESSION AND PRODUCTIVE TRANSLATION AT THE BEGINNING OF MOUSE DEVELOPMENT,   2002 01
  • Early development and gene expression of reconstructed embryos following fusion of bovine enucleated oocytes with somatic cells from different species,   2002 01
  • Application of an internal ribosomal entry sequence (IRES) bicistronic construct to investigate gene function in early mouse development,   2001 12
  • ANTISENSE INHIBITION OF LUCIFERASE EXPRESSION IN TRANSGENIC MOUSE PREIMPLANTATION EMBRYOS: A COMPARISON OF RNA AND OLIGODEOXYNUCLEOTIDES,   2001 01
  • Onset of gene expression in bovine embryos reconstructed with fibroblasts following reporter gene microinjection, 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)
  • 植物由来遺伝子導入マウスの作出とその解析 (2000年度日本繁殖生物学会シンポジウム講演論文), 松本 和也, Journal of Reproduction and Development,   2000 12
  • Microinjection of domestic and zoo animal spermatozoa into oocytes and their development, Hosoi Y, Torii R, Saeki K, Matsumoto K, Iritani A, Zygote,   1998 12

Misc

  • Examination of effective medium replacement interval for culture of rabbit immature oocytes, 和泉 広樹, 杉本 浩伸, 宮本 麻梨恵, 岸上 哲士, 松本 和也, 佐伯 和弘, 細井 美彦, Memoirs of Institute of Advanced Technology, Kinki University, 16, 27, 33,   2011 03 , http://ci.nii.ac.jp/naid/120005736481
    Summary:[要約] 哺乳動物の卵巣には、未成熟な卵母細胞が数多く存在する。これらを体外で培養し、成熟させる事ができれば、卵子資源として利用する事ができる。 本研究では、ウサギの未成熟卵母細胞を用いて体外発育培養(in vitro growth; IVG)の培養液交換の条件検討を行った。1996年にマウスのIVG において産仔作出に成功したEppig らの培養液の交換方法、2日に一回半量交換(half replacement /2 day; HR/2 day)に加え、毎日半量交換(half replacement /day;HR/day)、毎日全量交換(Full replacement /day; FR/day)の検討を行った。またIVG 後に体外成熟培養(in vitro maturation; IVM)、顕微授精(intracytoplasmic sperm injection; ICSI) を行った。その結果、IVG-IVM 後のM II期卵子への成熟率は他の区と比較してFR/day が高かった。前核形成率はHR/2 day と比較してHR/day は有意に高かったが、以降の発生率に差は見られなかった。また、IVG に供試した未成熟卵母細胞から胚盤胞期胚まで発生した割合は、FR/day において高かった。以上のことから、ウサギのIVG において培養液を毎日全量交換することが効果的であることが示された。 [Abstract] The mammalian ovary contains a large number of immature oocytes. If these oocytes can be cultured and obtain developmental competence, these oocytes are expected to be used as new resources for medicine. We studied medium replacement interval for in vitro growth(IVG)of rabbit immature oocytes. We employed the method of Eppig in 1996 that succeeded in production of mice and tried the half-volume replacement of medium every 2days(HR/2day). Additionally, We tried the half-volume replacement of medium(HR/day)and full-volume replacement of medium(FR/day)every day. After IVG, oocytes were cultured for in vitro maturation(IVM). Consequently, the percentage of oocytes that reached the metaphase II in FR/day was higher than that of others. After fertilization by intracytoplasmic sperm injection(ICSI), the percentage of oocytes that showed pronuclear formation in HR/day was significantly higher than that in HR/2 day, though there was no difference in developmental competence of reaching the cleavage stage and blastocyst stage for each condition. The developmental rate from the total number of oocytes used for IVG to the blastocyst stage was higher in FR/day. In conclusion, replacing the full volume of culture medium every day was found to be effective in rabbit IVG.
  • Somatic cell nuclear transfer with HDACI as a New Cloning Method, Satoshi Kishigami, Daisaku Iwamoto, Kazuhiro Saeki, Kazuya Matsumoto, Akira Iritani, Yoshihiko Hosoi, Animal Reproduction: New Research Developments, 329, 334,   2011 01 01 , https://www.scopus.com/inward/record.uri?partnerID=HzOxMe3b&scp=84892327982&origin=inward
    Summary:Somatic cell nuclear transfer (SCNT) has been diffused into a variety of researchfields including research for reprogramming and animal reproduction after Dolly thesheep, the first mammal to be cloned from an adult cell. However, since the birth ofDolly, the success rates have been inefficient in all the mammals examined as a result ofincomplete reprogramming of a somatic nucleus. The incomplete reprogramming hasbeen represented by abnormal epigenetic modifications, especially DNA methylation.Recently, we developed a new SCNT method using trichostatin A (TSA), a histonedeacetylase inhibitor (HDACi). This new SCNT with HDACi not only allows us toincrease their success rates 2-5 fold in F1 mice but also to clone outbred and inbredstrains which were "unclonable" strains before. Further, accumulating evidence suggeststhat the SCNT with HDACi can be applied to other species such as pig and bovine. Herewe introduce this new SCNT method and recent fruits. We also discuss the possiblemechanism underlying this dramatic improvement by HDACi. © 2009 Nova Science Publishers, Inc. All rights reserved.
  • The discussion on the target protein of histone deacetylase for reprogramming, Lee Ah Reum, Memoirs of Institute of Advanced Technology, Kinki University, 15, 1, 7,   2010 03 , http://ci.nii.ac.jp/naid/120005736379
    Summary:Somatic cell nuclear transfer technology enables somatic cells to achieve totipotency. In addition, pluripotency is acquired by introduction of only four transcriptional factors(Oct3/4, Sox2, Klf4, c-Myc) into somatic cells using iPS cell technology. However, one of the problems in these technologies is low efficiency of reprogramming. In previous studies, it was reported that the efficiency of reprogramming improved by histone deacetylase inhibitor (HDACi) treatment, but little is known of the molecular mechanism. The facilitation of histone acetylation by HDACi treatment relates to an increase in gene expression, and HDACi treatment also facilitates acetylation of non-histone proteins, and affects physiological activation. Thus, it is suggested that the acetylation of non-histone proteins relates to improvement of the reprogramming efficiency. In this review, we outline the mechanism of acetylation of non-histone proteins, and extract non-histone proteins possibly related to improving reprogramming fromGEO database.
  • Effect of cell cycle synchronization in rabbit somatic cell nuclear transfer, 歐 則克, 矢持 隆之, 木田 雄大, 中野 美穂, 岸上 哲士, 松本 和也, 佐伯 和弘, 入谷 明, 細井 美彦, Memoirs of Institute of Advanced Technology, Kinki University, 15, 37, 41,   2010 03 , http://ci.nii.ac.jp/naid/120005736383
    Summary:ウサギ体細胞クローンの作製において発生率の向上を目的に、核ドナーの細胞周期同期化処理を行い、ウサギ核移植後の胚発生に及ぼす影響を調べた。細胞周期同期化の影響を調べる為、対数増殖期、コンフルエント、血清飢餓培養を用いた。それぞれ胚盤胞期胚への発生率は3%(2/68)、2%(1/51) 12%(5/40)であり血清飢餓培養が最も有効であった。また発生速度を調べた結果、血清飢餓培養が他の区と比較して発生段階の速い胚が多く、脱出胚盤胞が観察された。以上の事より、血清飢餓培養により細胞周期をG0期にした細胞を核ドナーに用いることによってウサギ核移植胚の発生率の向上に有効であると示された。
  • The effect of electrical fusion method in rabbit somatic cell nuclear transfer, 木田 雄大, 矢持 隆之, 歐 則克, 中野 美穂, 岸上 哲士, 松本 和也, 佐伯 和弘, 入谷 明, 細井 美彦, Memoirs of Institute of Advanced Technology, Kinki University, 15, 53, 58,   2010 03 , http://ci.nii.ac.jp/naid/120005736385
    Summary:本実験では、核移植胚を作成する際の核移植法として主に用いられている注入法と電気融合法の二つの方法をウサギ体細胞核移植における核の移植法として用いて、どちらがウサギ体細胞核移植において有用であるかを検討した。その結果、注入法による核移植胚の卵割率、8 細胞期胚および胚盤胞期胚への発生率は、66%( 19/38)、31%( 9/38)、3%( 10/38)、電気融合法では、96%( 26/27)、89%( 24/27)、63%(17/27)で、電気融合法による核移植の方が注入法よりも胚盤胞期への発生率が有意に高かった(P<0.05)。また、胚の発生速度を比較したところ、電気融合法で得た胚の方が注入法で得た胚よりも発生速度が速くなることが示された。さらに、得られた胚盤胞期胚の細胞数をカウントしたが、有意な差は見られなかった。以上の結果より、ウサギ体細胞核移植胚の着床前発生において電気融合法の方が注入法よりも有用であることが示された。
  • Establish of embryonic stem cell derived from rabbit eggs fertilize by intra-cytoplasmic sperm injection, 中野 美穂, 杉本 浩伸, 荒田 隆志, 岸上 哲士, 松本 和也, 佐 伯 和弘, 細井 美彦, Memoirs of Institute of Advanced Technology, Kinki University, 15, 59, 65,   2010 03 , http://ci.nii.ac.jp/naid/120005736386
    Summary:胚性幹(Embryonic stem : ES)細胞は、体を構成するほぼすべての細胞へ分化することができる細胞である。そのため、ES細胞は再生医療の細胞移植材料として研究されてきた。細胞移植モデル動物は、飼育しやすく外科手術が容易で生理学的にヒトに近いことが求められる。そこで、ウサギは中型動物で大人しく室内飼育が可能なことから広くヒトモデル動物として研究に利用されてきた。さらに、マウスよりも体が大きく外科手術や術後の経過の観察も容易であることから、間葉系幹細胞を用いた目の角膜再生実験や骨形成促進実験、さらに軟骨細胞を用いた気管狭窄改善実験や関節軟骨再生実験などでヒトモデル動物として用いられてきた。これらのことから、ウサギES 細胞の樹立が可能となれば有用な細胞移植におけるヒトモデル動物及びシステムの構築になると期待できる。本実験ではウサギ卵子細胞質内精子注入法(Intra-cytoplasmic sperm injection : ICSI)により得られた胚からのES 細胞の樹立を試みた。樹立は、透明帯と栄養外胚葉を除去し内部細胞塊(Inner cell mass :ICM)をフィーダー細胞上へ播種し培養を行った。その結果、ES細胞様の初期コロニーが出現し未分化な遺伝子マーカーの発現も観察された。このことから、交配卵と同様に顕微受精胚からもES 細胞が樹立可能であることが示された。以上の事から、これらのES 細胞の樹立技術は、再生医療における細胞移植モデルのツールとして応用できると考えられる。
  • Proteome analysis of mouse preimplantation embryos, TOKORO Mikiko, KAWASUMI Miyuri, NAGAI Kohei, IKEGAMI Haruka, SATOH Manabu, SHIN Seung-Wook, NISHIKAWA Satoshi, HATANAKA Yuki, AMANO Tomoko, MITANI Tasuku, KATO Hiromi, ANZAI Masayuki, KISHIGAMI Satoshi, SAEKI Kazuhiro, HOSOI Yoshihiko, IRITANI Akira, MATSUMOTO Kazuya, Journal of mammalian ova research = 日本哺乳動物卵子学会誌, 26, 2,   2009 04 01 , http://ci.nii.ac.jp/naid/10024965056
  • The impact of EGTA as chelating calcium on oocyte activation in mice, 辻本 賀子, 岸上 哲士, 竹原 俊幸, 安齋 政幸, 松本 和也, 佐伯 和弘, 入谷 明, 細井 美彦, Memoirs of Institute of Advanced Technology, Kinki University, 14, 15, 20,   2009 03 , http://ci.nii.ac.jp/naid/120005736389
    Summary:人工的な卵子活性化法は、体細胞核移植や精子細胞を用いた産仔の作出に不可欠な技術である〔3〕。Sr<2+>を含んだ培養液は、受精の事象に似た反復的な細胞質内Ca^<2+>オシレーションを導き、マウス卵子における人工的な卵子活性化剤として広く使用されてきた。しかし、そのSr^<2+>が引き起こす卵子活性化は、Ca^<2+>を除いた培養液(Ca(-)培養液)で行われることが必要である。しかし、Ca(-)培養液を用いた場合、活性化に伴う形態異常や卵子の退行が起こるという報告もなされている。近年の研究から、金属イオンキレート剤であるグリコールエーテルジアミン四酢酸(Ethylene Glycol Tetraacetic Acid; EGTA〔9〕)を添加することにより、一般的なCa^<2+>含有培養液(Ca(+)培養液)においても、BDF1系統のマウスで効率的に活性化卵を作製できることや退行卵の頻度が低いことが明らかにされた。さらにこの活性化方法を用いたクローン胚の作出も報告されている〔1〕。しかし、BDF1系統マウス以外のマウス卵子での有用性は検証されておらず、またEGTAの添加の影響の詳細についても不明である。そこで本研究では、ICR系統マウス由来卵子を用いて、EGTAを添加したCa(+)とCa(-)培養液によって活性化を行い、活性化後の卵子の状態について比較・検討を行った。
  • Research about the development ability of in vitro grown rabbit oocytes, 宮本 有希, 是兼 真子, 杉本 浩伸, 竹原 俊幸, 伊藤 俊介, 岸上 哲士, 松本 和也, 佐伯 和弘, 入谷 明, 細井 美彦, Memoirs of Institute of Advanced Technology, Kinki University, 14, 31, 38,   2009 03 , http://ci.nii.ac.jp/naid/120005736391
    Summary:哺乳動物の卵巣内には排卵に至らない卵胞が数多く存在する。これらを体外で培養し、発生能力を有する卵子が得られれば、ヒト不妊治療等へ応用できると考えられる。体外発育培養(In vitro growth; IVG)は、卵形成、卵胞形成、卵巣の研究、稀少動物の保護、ヒト不妊治療等を目的として研究されてきた。受精能を有する卵子を得るためには小さく未熟な卵母細胞を発育させ、その後成熟させる必要がある。しかし、発育中の卵母細胞は種特異的なあるサイズに達するまで成熟できず、また種によってその発育期間が異なるためにマウス以外の動物種での研究は不十分であるのが現状である。IVGにおいて、卵子と周囲の細胞との相互作用を維持するために、卵子-顆粒膜細胞複合体 (Oocyte-Granulosa cell Complexes; OGCs)の立体構造の維持が重要となる。2004年、Hiraoらによってウシの体外発育培養培地にPolyvinylpyrrolidone (PVP) を添加することで立体構造が維持され、その後の発生率も向上することが報告された。そこで本実験では、ウサギ未成熟卵胞のIVGにおける培養条件の検討をおこない、ウサギ発育途上卵母細胞のIVG後の発生能力に関する検討をおこなった。その結果、ウサギIVGでは8日間の培養が好ましく、培地へのPVP添加がOGCsの立体構造を維持し、2%PVP添加が適している事が示唆された。またその後に、体外成熟培養 (In vitro maturation; IVM)、体外受精 (In vitro fertilization; IVF)をおこなった。IVGにより得られた卵子で前核形成が認められたことから、体外で発育させたウサギ卵母細胞が受精能を有していることが示された。Ito, Syunsuke
  • 飛騨牛由来白色脂肪組織の大規模プロテオーム解析, 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, 108th, 4,   2007 09 26 , http://jglobal.jst.go.jp/public/200902285597113656
  • Expression of transcription factor Cdx2 and Oct3/4 in mouse somatic nuclear transfer embryos., Memoirs of Institute of advanced Technology, Kinki University, 12, 33, 42,   2007 03
  • Development of an examination method of a methylation state of bull sperm DNA, Memoirs of Institute of Advanced Technology, Kinki University, 12, 43, 50,   2007 03
  • Localization of the autoimmune regulator (aire) protein in the gonads and embryonic stem cells in mice., Memoirs of Institute of Advanced Technology, Kinki University, 11, 15, 21,   2006 03
  • In vitro culture of CD9- and α6-integrin-expressingcells enriched by magnetic cell sorting from cryptorchid adult and pup testes in mice., Memoirs of Institute of Advanced technology, Kinki University, 11, 23, 34,   2006 03
  • Methylation og the 5'-upstream region og the H19 gene in mouse somatic cell, gametes, wild type and androgenetic ES cells., Memoirs of Institute of Advanced Technology, Kinki University, 11, 41, 49,   2006 03
  • Effects of sperm viability on kinetics of MPF activity of bovine oocytes following ICSI., 藤浪菜穂子, 細井美彦, 加藤博己, 松本和也, 佐伯和弘, 入谷明, 近畿大学先端技術総合研究所紀要, 9, 15, 20,   2004 02 28 , http://jglobal.jst.go.jp/public/200902257803909205
  • Correlation between follicle size and quality of oocytes from the superovulated cynomolgus monkey, Reproduction, Fertility and Development, 16, 1, 289,   2004 01
  • Application of transgenic rats for reproductive study, HORMONE FRONTIER IN GYNECOLOGY, 8, 1, 17, 23,   2001 , 招待有り
  • ウシ顕微授精はいの活性化による発生率について, 藤浪菜穂子, 細井美彦, 加藤博己, 松本和也, 佐伯和弘, 入谷明, 郁埼, 星合こう, 近畿大学生物理工学研究所紀要, 3, 39, 44,   2000 01 12 , http://jglobal.jst.go.jp/public/200902123812642924
  • Antisense inhibition of luciferase expression in transgenic mouse preimplantation embryos : a comparison of rna and oligodeoxynucleotides., Theriogenology, 55, 1, 413,   2001
  • Analysis of the role of egg integrins in sperm-egg binding and fusion, Y Takahashi, N Yamakawa, K Matsumoto, Y Toyoda, K Furukawa, E Sato, MOLECULAR REPRODUCTION AND DEVELOPMENT, 56, 3, 412, 423,   2000 07 , 10.1002/1098-2795(200007)56:3<412::AID-MRD12>3.0.CO;2-0
    Summary:Sperm-egg fusion is believed to be mediated via specific molecular interactions. Integrin alpha 6 beta 1 is a strong candidate for a sperm receptor on the egg plasma membrane. However, the ability of the egg integrin alpha 6 beta 1 to interact with molecules on intact sperm has not yet been proven. In this report, possible involvement of integrin alpha 6 beta 1 in sperm-egg interactions was examined by biochemical and immunocytochemical analyses. To identify egg molecules that specifically interact with sperm, we first incubated sperm with biotin-labeled egg surface proteins. Under this condition, solubilized proteins from eggs inhibited sperm-egg fusion. Western blot analysis under reducing conditions indicated that a major-labeled band of 135 kDa bound to sperm. An immunodepletion experiment using the anti-integrin alpha 6 antibody GoH3 indicated that the 135 kDa egg surface molecule that bound to sperm was the integrin alpha 6 subunit. To investigate the potential involvement of integrin alpha 6 beta 1 in sperm-egg fusion, we next examined the localization of integrin a6 and beta 1 subunits before and after fertilization by confocal laser microscopy. At an early stage of sperm-egg fusion, the integrin alpha 6 and beta 1 subunits were accumulated at the sperm binding site. The frequency of cluster formation was closely related to that of sperm-egg fusion, indicating that integrin receptors are accumulated by sperm destined for fusion. Taken together, these results strongly suggest that the integrin alpha 6 beta 1 is involved in sperm-egg binding leading to fusion via direct association of the integrin alpha 6 with sperm. Mol. Reprod. Dev. 56:412-423, 2000. (C) 2000 Wiley-Liss, Inc.
  • Transcriptional activity of follicle-stimulation hormone (FSH) receptor promoter in mouse ovarian granulosa cells., Theriogenology, 53, 406,   2000
  • Neuronal Apoptosis Inhibitory Protein (NAIP) May Enhance the Survival of Granulosa Cells which Indirectly Affects Oocyte Survival., indirectly affects oocyte survival. Mol. Reprod. Dev., 54, 103, 111,   1999 , 10.1002/(SICI)1098-2795(199910)54:2<103::AID-MRD1>3.0.CO;2-V
  • Effects of endogenous rat growth hormone gene expression suppressed by an antisense rna transgene on reproductive functions in transgenic rats., Mem. Res. Insti. BOST Kinki Uni., 3, 1, 9,   1999
  • Possible usage of the firefly luciferase gene as a gene expression marker in preimplantation embryos from transgenic mice., Mem. Res. Insti. BOST Kinki Uni., 2, 11, 18,   1999
  • A therapeutic role of prolactin sapplementation in ovarian sti mulatian for in vitro fertilization ; The bromocriptine-rebound method., J. Clin. Endocrinol. Metab., 82, 3603,   1997
  • Enhancement of number of evulations by administration of a pyrimidine compound with potentiating activity of angiogenesis by mice superovalated by PMSG and nCG, Kazuya MATSUMOTO, Seiki HARAGUCHI, Kazuchika MIYOSHI, Akira AWAYA, Eimei SATO, J. Reprod. Dev, 43, 2, 137, 141,   1997 , 10.1262/jrd.43.137
  • Synergistic effects of insulin-like growth factor II(IGF-II)with leukemia inhibiting factor(LIF)on establisment of rat plariopotential cell lines, Akio TAKAHASHI, Yumi TAKAHASHI, Kazuya MATSUMOTO, Kenji MIYATA, J. Vet. Med. Sci, 57, 3, 553, 556,   1995 , 10.1292/jvms.57.553
  • Evaluation of an artisense RNA trnsgene for inhibiting growth hormone gene expression in transgenic rats, Dev. Gene., 16, 3, 273, 277,   1995 , 10.1002/dvg.1020160307
  • Oneset of paternal gene actination in early mouse embryos fertilized with transgenic merse speim, Mol. Reprod. Dev., 39, 136,   1994
  • Feasibility of firefly lucifuase gene as in situ gene expression marker in alive preimplantation embryos from transgenic mice, Theriogenology,   1994
  • Growth retardation in rats whose growth hormone gene expression was suppressed by antisense RNA transgene, 36, 53,   1993
  • Production of identical twins by separating two-cell rat embryos., Gamete Res., 22, 257, 263,   1989
  • Effective production of monozygotic identical goat twin from 3 split embryos., Mem. Coll. Agric. Kyoto Univ., 131, 1, 6,   1988

Research Grants & Projects

  • Study on programmed cell death in germ cells.
  • Study on the developmental regulation of mammalian preimplantation embryo