KINDAI UNIVERSITY


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ANZAI Masayuki

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FacultyInstitute of Advanced Technology / Graduate School of Biology-Oriented Science and Technology
PositionAssociate Professor
Degree
Commentator Guidehttps://www.kindai.ac.jp/meikan/502-anzai-masayuki.html
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Last Updated :2020/11/25

Education and Career

Academic & Professional Experience

  •   2014 04 ,  - 現在, Institute of Advanced Technology, Kindai University
  •   2011 04 ,  - 2014 03 , Institute of Advanced Technology, Kindai University

Research Activities

Research Areas

  • Life sciences, Laboratory animal science

Research Interests

  • RNAi

Published Papers

  • 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
  • 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.
  • Study on fertilization and embryo culture in without albumin HTF medium from using L-Carnitine., 16, 43 - 50, Mar. 2011 , Refereed
  • Establishment of in situ hybridization technique with mouse single ovum that uses QuantiGene ViewRNA, 16, 35 - 42, Mar. 2011 , Refereed
  • Short term storage of Mouse epididymal spermatozoa by Antifreeze protein addition at cold temperature, 16, 51 - 58, Mar. 2011 , Refereed
  • Investigation of zona drilled by laser anf in vitro fertilization of oocytes that matured in vitro with C57BL/6 strain, 15, 27 - 36, Mar. 2010 , Refereed
  • Short term strage of Wistar rat epididymal spermatozoa at cold temperature, 15, 17 - 26, Mar. 2010 , Refereed
  • The develpoment capacity of oocytes that matured in vitro with C57BL/6 MICE, 44(2), 43 - 48, Dec. 2009 , Refereed
  • Study of fertilization and developmant of Wistar rat eggs in vitro, 147, 1 - 8, Mar. 2009
  • In vitro fertilization using cryopreserved laser-microdissected oocytes on the inbred mouse strains, MIYAJI Shiori, ANZAI Masayuki, KODA Yuna, YANAGI Miho, NAKASHIMA Tatsuyuki, KAWABE Toshiaki, KANEKO Takehito, NAKAGATA Naomi, 43(1), 25 - 29, Jun. 2008 , Refereed
  • Expression of transcription factor Cdx2 and Oct3/4 in mouse somatic nuclear transfer embryos., Memoirs of Institute of advanced Technology, Kinki University, Memoirs of Institute of advanced Technology, Kinki University, 12, 33 - 42, Mar. 2007
  • Identification and characterization of the 5 '-flanking region of three mouse maternal genes (Histone H1oo, Nucleoplasmin 2, and Zygote arrest 1): Transcriptional activity in mouse oocytes, K. Tsunemoto, K. Matsumoto, M. Anzai, M. Hayakumo, T. Amano, T. Mitani, H. Kato, Y. Hosoi, K. Saeki, A. Iritani, REPRODUCTION FERTILITY AND DEVELOPMENT, REPRODUCTION FERTILITY AND DEVELOPMENT, 19(1), 257 - 258, 2007 , Refereed
  • Methylation og the 5'-upstream region og the H19 gene in mouse somatic cell, gametes, wild type and androgenetic ES cells., 11, 41 - 49, Mar. 2006
  • Investigation of in vitro fertilization and early embryonic vitrification in JAX | MICE C57BL/6J, ANZAI Masayuki, KASUYA Yoshie, HOSOI Yoshihiko, MATSUMOTO Kazuya, SAEKI Kazuhiro, IRITANI Akira, 40(2), 87 - 90, Dec. 2005 , Refereed
  • Methylation of the 5’-upstream region of the H19 gene in mouse somatic cell, gametes, wild type and androgenetic ES cells, Reproduction, Fertility and Development, Reproduction, Fertility and Development, 17(1,2), 261 - 262, Jan. 2005
  • Analysis of DNA Methylation Pattern in Mouse Early Embryos by Bisulfite-Sequencing Method, Yuichi Unno, Miyuri Kawasumi, Kazuya Matsumoto, Tomoko Amano, Kazuhiro Saeki, Yoshihiko Hosoi, Akira Iritani, Hiromi Kato, Kazuya Matsumoto, Masayuki Anzai, Tasuku Mitani, Kazuhiro Saeki, Yoshihiko Hosoi, Akira Iritani, Journal of Mammalian Ova Research, Journal of Mammalian Ova Research, 22(4), 241 - 245, 2005 , Refereed
    Summary:Recently, with the acetylation of histone and the modification of chromatin structure, the methylation of cytosine residue within CpG dinucleotides in genomic DNA sequence attracts many researchers' attention as one of major epigenetic regulation systems of gene expression. There are several methods (immunofluorescence with 5-methylcytosine specific antibody and methylationsensitive restriction enzyme-PCR method, etc.) to analyze the methylation of cytosine residue. Bisulfitesequencing method, which is one of methods for analyzing the methylation of cytosine residue in genomic DNA sequence, has advantages of high sensitivity and analyzing the methylation of cytosine residue directly. In this note, the detailed procedure of bisulfite-sequencing method for mouse early Preimplantation embryos is described. © 2005, JAPANESE SOCIETY OF OVA RESEARCH. All rights reserved.
  • [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
  • Developmental competence of interspecies cloned embryos produced using cells from large Japanese field mice (Apodemus speciosus) and oocytes from laboratory mice (Mus musculus domesticus)., Rika Azuma, Yuki Hatanaka, Seung-Wook Shin, Hitoshi Murai, Minoru Miyashita, Masayuki Anzai, Kazuya Matsumoto, The Journal of reproduction and development, The Journal of reproduction and development, 66(3), 255 - 263, Jun. 12 2020 , Refereed
    Summary:The large Japanese field mouse (Apodemus speciosus) is endemic to Japan and may be used as an animal model for studies related to environmental pollution, medical science, and basic biology. However, the large Japanese field mouse has low reproductive ability due to the small number of oocytes ovulated per female. To produce experimental models, we investigated the in vitro developmental potential of interspecies somatic cell nuclear transfer (iSCNT) embryos produced by fusing tail tip cells from the large Japanese field mouse with enucleated oocytes from laboratory mice (Mus musculus domesticus). Only a small number of iSCNT embryos developed to the 4-cell (0-4%) and blastocysts (0-1%) stages under sequential treatment using trichostatin A (TSA) and vitamin C (VC) supplemented with deionized bovine serum albumin (d-BSA). This sequential treatment led to the reduction in H3K9 trimethylation and did not affect H3K4 trimethylation in at least the 2-cell stage of the iSCNT embryos. Moreover, iSCNT embryos that received tail tip cells with exposure treatment to ooplasm from cell fusion to oocyte activation or VC treatment prior to cell fusion did not exhibit significant in vitro development improvement compared to that of each control group. This suggests that large Japanese field mice/laboratory mice iSCNT embryos that received sequential treatment using TSA and VC with d-BSA may have slightly better developmental potential beyond the 4-cell stage. Our results provide insights into the reprogramming barriers impeding the wider implementation of iSCNT technology.
  • Development of feline embryos produced by Piezo-actuated intracytoplasmic sperm injection of elongated spermatids., Yasunori Tsujimoto, Kana Fujiki, Md Emtiaj Alam, Masaya Tsukamoto, Rika Azuma, Ryoji Kanegi, Masayuki Anzai, Toshio Inaba, Kikuya Sugiura, Shingo Hatoya, The Journal of reproduction and development, The Journal of reproduction and development, 65(3), 245 - 250, Jun. 14 2019 , Refereed
    Summary:Piezo-actuated intracytoplasmic sperm injection (Piezo-ICSI) is used as an efficient in vitro fertilization method with various animals. With this method, elongated spermatids are collected from testicular tissues and are easier to obtain from animals that unexpectedly die than ejaculate sperm. Additionally, elongated spermatid injection often results in the development of embryos and offspring. To develop assisted reproductive techniques (ARTs) for domestic cats, we examined the effects of oocyte activation on cleavage and embryo development after Piezo-ICSI with motile sperm (experiment 1) and after Piezo-ICSI with either testicular sperm or elongated spermatids (experiment 2). In experiment 1, the proportions of cleaved embryos, morulas, and blastocysts following Piezo-ICSI with ethanol activation were significantly higher (P < 0.05) than in the non-activated groups. However, the proportion of blastocysts and the blastocyst quality did not differ significantly (P > 0.05) between the ethanol-activated and non-activated groups. In experiment 2, the cleavage frequencies of oocytes after Piezo-ICSI of testicular sperm or elongated spermatids and ethanol activation were higher (P < 0.05) than that of oocytes in the non-activated group, but the occurrence of blastocyst formation and quality of blastocysts did not differ between the activated and non-activated groups. In summary, cat embryos can be produced by Piezo-actuated microinjection of elongated spermatids. Ethanol activation increased the frequency of cleavage, but it affected neither the occurrence of blastocyst development nor the quality of blastocysts. These results represent an expansion in the repertoire of ARTs that are potentially applicable to both domestic and endangered species of cats.
  • Combinational Treatment of Trichostatin A and Vitamin C Improves the Efficiency of Cloning Mice by Somatic Cell Nuclear Transfer., Rika Azuma, Kei Miyamoto, Mami Oikawa, Masayasu Yamada, Masayuki Anzai, Journal of visualized experiments : JoVE, Journal of visualized experiments : JoVE, (134), Apr. 26 2018 , Refereed
    Summary:Somatic cell nuclear transfer (SCNT) provides a unique opportunity to directly produce a cloned animal from a donor cell, and it requires the use of skillful techniques. Additionally, the efficiencies of cloning have remained low since the successful production of cloned animals, especially mice. There have been many attempts to improve the cloning efficiency, and trichostatin A (TSA), a histone deacetylase inhibitor, has been widely used to enhance the efficiency of cloning. Here, we report a dramatically improved cloning method in mice. This somatic cell nuclear transfer method involves usage of Hemagglutinating virus of Japan Envelope (HVJ-E), which enables easy manipulation. Moreover, the treatment using two small molecules, TSA and vitamin C (VC), with deionized bovine serum albumin (dBSA), is highly effective for embryonic development. This approach requires neither additional injection nor genetic manipulation, and thus presents a simple, suitable method for practical use. This method could become a technically feasible approach for researchers to produce genetically modified animals from cultured cells. Furthermore, it might be a useful way for the rescue of endangered animals via cloning.
  • 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.
  • 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.
  • Cryopreservation of a small number of human sperm using enzymatically fabricated, hollow hyaluronan microcapsules handled by conventional ICSI procedures, Kazuhisa Tomita, Shinji Sakai, Mehdi Khanmohammadi, Takayuki Yamochi, Shu Hashimoto, Masayuki Anzai, Yoshiharu Morimoto, Masahito Taya, Yoshihiko Hosoi, Journal of Assisted Reproduction and Genetics, Journal of Assisted Reproduction and Genetics, 33(4), 501 - 511, Apr. 2016 , Refereed
    Summary:© 2016, Springer Science+Business Media New York. Purpose: We investigated whether enzymatically fabricated hyaluronan (HA) microcapsules were feasible for use in the cryopreservation of a small number of sperm. Methods: HA microcapsules were fabricated using a system of water-immiscible fluid under laminar flow. Three sperm were injected into a hollow HA microcapsule using a micromanipulator. Capsules containing injected sperm were incubated in a freezing medium composed of sucrose as the cryoprotectant and then placed in a Cryotop® device and plunged into liquid nitrogen. After thawing, the capsule was degraded by hyaluronidase, and the recovery rate of sperm and their motility were investigated. Results: The HA microcapsule measuring 200 μm in diameter and with a 30-μm thick membrane was handled using a conventional intracytoplasmic sperm injection (ICSI) system, and the procedure involved the injection of sperm into the capsule. The HA microcapsules containing sperm were cryopreserved in a Cryotop® device and decomposed by the addition of hyaluronidase. The recovery rate of sperm after cryopreservation and degradation of HA microcapsules was sufficient for use in clinical practice (90 %). Conclusions: Hollow HA microcapsules can be used for the cryopreservation of a small number of sperm without producing adverse effects on sperm quality.
  • 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.
  • 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.
  • The influence of reduced glutathione in fertilization medium on the fertility of in vitro-matured C57BL/6 mouse oocytes, Y. Ishizuka, M. Nishimura, K. Matsumoto, M. Miyashita, T. Takeo, N. Nakagata, Y. Hosoi, M. Anzai, THERIOGENOLOGY, THERIOGENOLOGY, 80(5), 421 - 426, Sep. 2013 , Refereed
    Summary:It is well known that IVM oocytes show a decreased potential for fertility and development compared with in vivo-matured oocytes. In this study, we added reduced glutathione (GSH) to the fertilization medium during IVF to investigate its effect on the fertility and early embryo development of IVM oocytes. The fertilization rate for IVM oocytes and fresh sperm increased with the addition of GSH (0, 1.0, and 2.0 mM: 51%, 76%, and 70%). Moreover, the addition of GSH to the fertilization medium also improved the developmental potential compared with the control sample (0 mM). In addition, we performed IVF using IVM oocytes and frozen/thawed sperm that had been cryopreserved in a mouse bank. Results indicated a marked increase in the fertilization rate when 1.0 mM GSH was added to the fertilization medium compared with when no GSM was used (0.0 mM GSH: 2% (3/195); 1.0 mM GSH: 33% (156/468)). Furthermore, the fertilization rate improved dramatically via zona drilling using laser equipment (52%; 267/516), whereas normal offspring were obtainsed after transferring embryos created via IVF using IVM oocytes and frozen/thawed sperm. This is the first report in which offspring have been obtained via IVF using IVM oocytes and frozen/thawed sperm. (c) 2013 Elsevier Inc. All rights reserved.
  • 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.
  • 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.
  • EFFECT OF Dnmt1p mRNA KNOCK DOWN ON Dnmt1 PROTEIN TRANSLATION IN MOUSE TESTIS, H. Kato, R. Kitamura, H. Yamaguchi, Y. Numata, T. Kijima, M. Anzai, T. Mitani, K. Matsumoto, K. Saeki, Y. Hosoi, A. Iritani, REPRODUCTION FERTILITY AND DEVELOPMENT, REPRODUCTION FERTILITY AND DEVELOPMENT, 24(1), 202 - 202, 2012 , Refereed
  • In Vivo Application of an RNAi Strategy for the Selective Suppression of a Mutant Allele, Takayuki Kubodera, Hiromi Yamada, Masayuki Anzai, Shinga Ohira, Shigefumi Yokota, Yukihiko Hirai, Hideki Mochizuki, Takashi Shimada, Tasuku Mitani, Hidehiro Mizusawa, Takanori Yokota, HUMAN GENE THERAPY, HUMAN GENE THERAPY, 22(1), 27 - 34, Jan. 2011 , Refereed
    Summary:Gene therapy for dominantly inherited diseases with small interfering RNA (siRNA) requires mutant allele-specific suppression when genes in which mutation causes disease normally have an important role. We previously proposed a strategy for selective suppression of mutant alleles; both mutant and wild-type alleles are inhibited by most effective siRNA, and wild-type protein is restored using mRNA mutated to be resistant to the siRNA. Here, to prove the principle of this strategy in vivo, we applied it to our previously reported anti-copper/zinc superoxide dismutase (SOD1) short hairpin RNA (shRNA) transgenic (Tg) mice, in which the expression of the endogenous wild-type SOD1 gene was inhibited by more than 80%. These shRNA Tg mice showed hepatic lipid accumulation with mild liver dysfunction due to downregulation of endogenous wild-type SOD1. To rescue this side effect, we generated siRNA-resistant SOD1 Tg mice and crossed them with anti-SOD1 shRNA Tg mice, resulting in the disappearance of lipid accumulation in the liver. Furthermore, we also succeeded in mutant SOD1-specific gene suppression in the liver of SOD1(G93A) Tg mice, a model for amyotrophic lateral sclerosis, using intravenously administered viral vectors. Our method may prove useful for siRNA-based gene therapy for dominantly inherited diseases.
  • 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.
  • Differentiation diversity of mouse parthenogenetic embryonic stem cells in chimeric mice, Yuta Onodera, Takeshi Teramura, Madoka Ozawa, Toshiyuki Takehara, Tasuku Mitani, Masayuki Anzai, Norimasa Sagawa, Chiaki Hamanishi, Yoshihiko Hosoi, Kanji Fukuda, THERIOGENOLOGY, THERIOGENOLOGY, 74(1), 135 - 145, Jul. 2010 , Refereed
    Summary:Recent studies have illustrated multiple differentiation potentials of embryonic stem cells (ESCs), derived from parthenogenetic embryos, to various kinds of cells (all three embryonic germ layers). However, differentiation diversity of the parthenogenetic ESCs (PgESCs) in vivo remains to be elucidated. In the present study, we established mouse PgESC-lines and observed their contribution diversity in vivo by producing chimeric mice using embryos possessing single nucleotide polymorphisms of mitochondrial DNA (mtDNA) as hosts. Based on southern blot analysis using specific probes to detect the SNPs on mtDNA, PgESC-derived mtDNA were contained in many organs such as brain, lung, and heart of the chimeric mouse. We concluded that PgESCs contributed to various internal organs in vivo, and that they were also stably maintained in adult animals. (C) 2010 Elsevier Inc. All rights reserved.
  • Short-Term Storage and Transport at Cold Temperatures of 2-Cell Mouse Embryos Produced by Cryopreserved Sperm, Toru Takeo, Tomoko Kondo, Yukie Haruguchi, Kiyoko Fukumoto, Yoshiko Nakagawa, Yumi Takeshita, Yuko Nakamuta, Shuuji Tsuchiyama, Norihiko Shimizu, Takanori Hasegawa, Motohito Goto, Hitoshi Miyachi, Masayuki Anzai, Rie Fujikawa, Koji Nomaru, Takehito Kaneko, Yoshiaki Itagaki, Naomi Nakagata, JOURNAL OF THE AMERICAN ASSOCIATION FOR LABORATORY ANIMAL SCIENCE, JOURNAL OF THE AMERICAN ASSOCIATION FOR LABORATORY ANIMAL SCIENCE, 49(4), 415 - 419, Jul. 2010 , Refereed
    Summary:At refrigerated temperatures, mouse embryos can maintain developmental ability for short periods. Previously, we succeeded in transporting vitrified and warmed 2-cell mouse embryos while maintaining developmental ability at refrigerated temperatures for 50 h. Transport of nonfrozen embryos is an easier and more useful means of exchanging genetically engineered mice between laboratories than is transport of cryopreserved embryos. Here we examined the developmental ability of transported 2-cell embryos that were produced through in vitro fertilization using cryopreserved sperm. Results show that 2-cell embryos produced by cryopreserved sperm can develop into blastocysts after cold storage for 24, 48, and 72 h. Transported 2-cell embryos produced by cryopreserved sperm yielded a favorable number of pups in all of the receiving laboratories after transport lasting 48 to 52 h. In summary, cold storage and transport of 2-cell embryos derived from cryopreserved sperm at refrigerated temperatures provides a novel means of transporting genetically engineered mice as an alternative to the transport of cryopreserved embryos and sperm.
  • FIBROBLAST GROWTH FACTOR 4 PROMOTES THE DEVELOPMENT OF SOMATIC CELL NUCLEAR TRANSFER EMBRYOS IN MICE, T. Mitani, M. Morita, M. Anzai, Y. Nishiyama, K. Moriki, H. Kawamura, H. Kato, K. Saeki, Y. Hosoi, A. Iritani, REPRODUCTION FERTILITY AND DEVELOPMENT, REPRODUCTION FERTILITY AND DEVELOPMENT, 22(1), 193 - 194, 2010 , Refereed
  • RECOVERY OF CELL NUCLEI FROM 15 000-YEAR-OLD MAMMOTH TISSUES AND INJECTION INTO MOUSE ENUCLEATED MATURED OOCYTES, H. Kato, M. Anzai, T. Mitani, M. Morita, Y. Nishiyama, A. Nakao, K. Kondo, P. A. Lazarev, T. Ohtani, Y. Shibata, A. Iritani, REPRODUCTION FERTILITY AND DEVELOPMENT, REPRODUCTION FERTILITY AND DEVELOPMENT, 22(1), 189 - 189, 2010 , Refereed
  • Recovery of cell nuclei from 15,000 years old mammoth tissues and its injection into mouse enucleated matured oocytes, Hiromi Kato, Masayuki Anzai, Tasuku Mitani, Masahiro Morita, Yui Nishiyama, Akemi Nakao, Kenji Kondo, Petr A. Lazarev, Tsuyoshi Ohtani, Yasuyuki Shibata, Akira Iritani, PROCEEDINGS OF THE JAPAN ACADEMY SERIES B-PHYSICAL AND BIOLOGICAL SCIENCES, PROCEEDINGS OF THE JAPAN ACADEMY SERIES B-PHYSICAL AND BIOLOGICAL SCIENCES, 85(7), 240 - 247, Jul. 2009 , Refereed
    Summary:Here, we report the recovery of cell nuclei front 14,000-15,000 years old mammoth tissues and the injection of those nuclei into mouse enucleated matured oocytes by somatic cell nuclear transfer (SCNT). From both skin and muscle tissues, cell nucleus-like structures were successfully recovered. Those nuclei were then injected into enucleated oocytes and more than half of the oocytes were able to survive. Injected nuclei were not taken apart and remained its nuclear structure. Those oocytes did not show disappearance of nuclear membrane or premature chromosome condensation (PCC) at 1 hour after injection and did not form pronuclear-like structures at 7 hours after injection. As half of the oocytes injected with nuclei derived from frozen-thawed mouse bone marrow cells were able to form pronuclear-like structures, it might be possible to promote the cell cycle of nuclei from ancient animal tissues by suitable pre-treatment in SCNT. This is the first report of SCNT with nuclei derived from mammoth tissues.
  • 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.
  • Birth of mice from vitrified/warmed 2-cell embryos transported at a cold temperature, Toru Takeo, Takehito Kaneko, Yukie Haruguchi, Kiyoko Fukumoto, Hiromi Machida, Mika Koga, Yoshiko Nakagawa, Yumi Takeshita, Toyokazu Matsuguma, Shuuji Tsuchiyama, Norihiko Shimizu, Takanori Hasegawa, Motohito Goto, Hitoshi Miyachi, Masayuki Anzai, Ena Nakatsukasa, Koji Nomaru, Naomi Nakagata, CRYOBIOLOGY, CRYOBIOLOGY, 58(2), 196 - 202, Apr. 2009 , Refereed
    Summary:Cryopreservation of 2-cell embryos is an effective technology for storage of genetically engineered mouse strains. Transport of genetically engineered mice between laboratories has frequently been performed using such cryopreserved 2-cell embryos. However, the receiving laboratory requires proficient skills and special instruments to obtain live young from cryopreserved and transported embryos. Therefore, in this study, we tried to address the storage and transport of vitrified/warmed 2-cell embryos at a cold temperature. In cold storage experiments, the development rates of 2-cell embryos stored in M2 medium for 24,48 and 72 h into blastocysts were relatively high (83%, 63% and 43%, respectively). Although, 2-cell embryos stored in PB1 and mWM maintained the developmental potency for 24 h, the rates were markedly decreased to low levels after 48 h (PB1: 0%; mWM: 5%). In transport experiments, many pups were obtained from vitrified/warmed 2-cell embryos transported at a cold temperature in all receiving laboratories (incidence of successful development: 49%; 249/511). In summary, short-term storage and transport of vitrified/warmed 2-cell embryos in M2 medium at a cold temperature can maintain their ability to develop into live young. (C) 2008 Elsevier Inc. All rights reserved.
  • 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.
  • Silencing efficiency differs among tissues and endogenous microRNA pathway is preserved in short hairpin RNA transgenic mice, Hiroki Sasaguri, Tasuku Mitani, Masayuki Anzai, Takayuki Kubodera, Yuki Saito, Hiromi Yamada, Hidehiro Mizusawa, Takanori Yokota, FEBS LETTERS, FEBS LETTERS, 583(1), 213 - 218, Jan. 2009 , Refereed
    Summary:In short hairpin RNA (shRNA) transgenic mice, the tissue difference in gene silencing efficiency and oversaturation of microRNA (miRNA) pathway have not been well assessed. We studied these problems in our previously-reported anti-copper/zinc superoxide dismutase (SOD1) shRNA transgenic mice. Although there was a tissue difference (liver and skeletal muscle, >95%; central nervous system and lung, similar to 80%), the target gene silencing was systemic and our anti-SOD1 shRNA transgenic mice recapitulated the SOD1-null mice. Neither endogenous miRNAs nor their target gene levels were altered, indicating the preservation of endogenous miRNA pathways. We think that the shRNA transgenic mice can be utilized for gene analysis. (C) 2008 Federation of European Biochemical Societies. Published by Elsevier B. V. All rights reserved.
  • 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.
  • Expression of transcription factors specific to the trophoblast lineage in mouse somatic nuclear transfer embryos, T. Mitani, M. Nishiwaki, M. Anzai, H. Kato, Y. Hosoi, A. Iritani, REPRODUCTION FERTILITY AND DEVELOPMENT, REPRODUCTION FERTILITY AND DEVELOPMENT, 20(1), 103 - 104, 2008 , Refereed
  • Early development of reconstructed mouse embryos using bone marrow cells frozen without cryoprotectant, H. Kato, A. Nakao, M. Nishiwaki, M. Anzai, T. Mitani, K. Matsumoto, K. Saeki, Y. Hosoi, A. Iritani, REPRODUCTION FERTILITY AND DEVELOPMENT, REPRODUCTION FERTILITY AND DEVELOPMENT, 20(1), 100 - 100, 2008 , Refereed
  • 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.
  • DNA methylation profiles of upstream elements of Oct-3/4 gene in in vitro fertilization (IVF) and somatic cell nuclear-transferred (SCNT) embryos, M. Kawasumi, Y. Unno, M. Nishiwaki, K. Matsumoto, M. Anzai, T. Amano, T. Mitani, H. Kato, K. Saeki, Y. Hosoi, A. Iritani, REPRODUCTION FERTILITY AND DEVELOPMENT, REPRODUCTION FERTILITY AND DEVELOPMENT, 19(1), 143 - 143, 2007 , Refereed
  • Increase of disease duration of amyotrophic lateral sclerosis in a mouse model by transgenic small interfering RNA, Takanori Yokota, Hiroki Sasaguri, Yuki Saito, Hiromi Yamada, Toshinori Unno, Yuki Yamamoto, Takayuki Kubodera, Masayuki Anzai, Tasuku Mitani, Hidehiro Mizusawa, ARCHIVES OF NEUROLOGY, ARCHIVES OF NEUROLOGY, 64(1), 145 - 146, Jan. 2007 , Refereed
  • 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.
  • Transgenic small interfering RNA halts amyotrophic lateral sclerosis in a mouse model, Y Saito, T Yokota, T Mitani, K Ito, M Anzai, M Miyagishi, K Taira, H Mizusawa, JOURNAL OF BIOLOGICAL CHEMISTRY, JOURNAL OF BIOLOGICAL CHEMISTRY, 280(52), 42826 - 42830, Dec. 2005 , Refereed
    Summary:Many autosomal dominant diseases such as familial amyotrophic lateral sclerosis (ALS) with copper/zinc superoxide dismutase (SOD1) mutation may be induced by missense point mutations that result in the production of proteins with toxic properties. Reduction in the encoding of proteins from such mutated genes can therefore be expected to improve the disease phenotype. The duplex of 21-nucleotide RNA, known as small interfering RNA (siRNA), has recently emerged as a powerful gene silencing tool. We made transgenic (Tg) mice with modified siRNA, which had multiple mismatch alternations within the sense strand, to prevent the "shutdown phenomenon" of transgenic siRNA. Consequently, the in vivo knockdown effect of siRNA on SOD1 expression did not diminish over four generations. When we crossed these anti-SOD1 siRNA Tg mice with SOD1(G93A) Tg mice, a model for ALS, siRNA prevented the development of disease by inhibiting mutant G93A SOD1 production in the central nervous system. Our findings clearly proved the principle that siRNA-mediated gene silencing can stop the development of familial ALS with SOD1 mutation.
  • The natural history of maternal immunization against foetal platelet alloantigens, Hitoshi Ohto, S. Miura, H. Ariga, T. Ishii, K. Fujimori, S. Morita, C. Takeuchi, Y. Tohyama, T. Yamaguchi, M. Anzai, N. Ujiie, T. Okawa, A. Sato, H. Suzuki, T. Okada, A. Ishijima, S. Inoue, T. Enomoto, K. Hando, S. Imamura, M. Sugafuji, K. Takasaki, K. Ohsugi, S. Kim, Transfusion Medicine, Transfusion Medicine, 14(6), 399 - 408, Dec. 2004 , Refereed
    Summary:Foetomaternal alloimmune thrombocytopenia (FMAIT) occurs when maternal antibodies of an antigen-negative mother cause destruction of sensitized foetal platelets. In Caucasian populations, 6-12% of human platelet antigen (HPA)-1a-negative women develop anti-HPA-1a, and the incidence of clinically affected cases is estimated to be 10-20% of immunized women. This study was performed in order to elucidate the rate of maternal immunization, incidence of FMAIT and the likely outcome of the condition in Asians. Excluding two or more pregnancies during the period, serum samples from 24630 pregnant women, mainly Japanese, were screened for antibodies against platelet alloantigens by means of mixed passive haemagglutination (MPHA) (Anti-HPA-MPHA, Olympus, Tokyo). Antibodies were detected in 0-91% (223/24630) of the women's samples and the immunization rate was correlated with the number of pregnancies. Antibody specificity included anti-HPA-4b (49), anti-HPA-5a (three), anti-HPA-5b (168), anti-HPA-4b + 5b (one) and anti-Naka (CD36) (two). No alloimmunization was observed within the HPÁ-1, HPA-2, HPA-3 or HPA-6 systems. Among HPA-4b- or HPA-5b-negative women, 24% or 14% estimated, respectively, had antibodies and 26% (10/38) or 10% (12/125) of neonates, respectively, born to these mothers developed thrombocytopenia. Two neonates born to mothers having anti-HPA-4b developed generalized purpura. No cases of intracranial bleeding or death due to FMAIT were recorded. Generalized purpura due to FMAIT occurs in one in 9359 (95% CI: 1 in 77519-1 in 2591) pregnancies solely because of HPA-4b incompatibility.
  • Reversible suppression of glutamatergic neurotransmission of cerebellar granule cells in vivo by genetically manipulated expression of tetanus neurotoxin light chain, M Yamamoto, N Wada, Y Kitabatake, D Watanabe, M Anzai, M Yokoyama, Y Teranishi, S Nakanishi, JOURNAL OF NEUROSCIENCE, JOURNAL OF NEUROSCIENCE, 23(17), 6759 - 6767, Jul. 2003 , Refereed
    Summary:We developed a novel technique that allowed reversible suppression of glutamatergic neurotransmission in the cerebellar network. We generated two lines of transgenic mice termed Tet and TeNT mice and crossed the two transgenic lines to produce the Tet/TeNT double transgenic mice. In the Tet mice, the tetracycline-controlled reverse activator (rtTA) was expressed selectively in cerebellar granule cells by the promoter function of the GABA(A) receptor alpha6 subunit gene. In the TeNT mice, the fusion gene of tetanus neurotoxin light chain (TeNT) and enhanced green fluorescent protein (EGFP) was designed to be induced by the interaction of doxycycline (DOX)-activated rtTA with the tetracycline-responsive promoter. The Tet/TeNT mice grew normally even after DOX treatment and exhibited a restricted DOX-dependent expression of TeNT in cerebellar granule cells. Along with this expression, TeNT proteolytically cleaved the synaptic vesicle protein VAMP2 (also termed synaptobrevin2) and reduced glutamate release from granule cells. Both cleavage of VAMP2/synaptobrevin2 and reduction of glutamate release were reversed by removal of DOX. Among the four genotypes generated by heterozygous crossing of Tet and TeNT mice, only Tet/TeNT mice showed DOX-dependent reversible motor impairments as analyzed with fixed bar and rota-rod tests. Reversible suppression of glutamatergic neurotransmission thus can be manipulated with spatiotemporal accuracy by DOX treatment and removal. These transgenic mice will serve as an animal model to study the cerebellar function in motor coordination and learning.
  • Transport of wild mice genetic material by in vitro fertilization, cryopreservation, and embryo transfer, H Suzuki, N Nakagata, M Anzai, K Tsuchiya, M Nakura, S Yamaguchi, Y Toyoda, LABORATORY ANIMAL SCIENCE, LABORATORY ANIMAL SCIENCE, 46(6), 687 - 688, Dec. 1996 , 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.
  • Survey of Hanganutziu and Deicher antibodies in operated patients, T. Higashihara, T. Takeshima, M. Anzai, M. Tomioka, K. Matsumoto, K. Y. Nishida Kitamura, K. Okinaga, M. Naiki, International Archives of Allergy and Applied Immunology, International Archives of Allergy and Applied Immunology, 95(2-3), 231 - 235, 1991 , Refereed
    Summary:The appearance of Hanganutziu and Deicher (HD) antibody in the sera of patients suffering from various diseases, including malignancies of some organs and liver disorders, was investigated by enzyme-linked immunosorbent assay using N-glycolylneuraminyl-lactosylceramide (HD3) and 4-O-acetyl-HD3 as the antigenic molecules. More than 25% of sera from patients suffering from malignancies, cholelithiasis and liver cirrhosis had HD antibody, whereas none of 41 sera from healthy persons had HD antibody. The percentage of HD antibody-positive patients was similar in stages I, II and III of gastric cancer and recurrence cases. Antibody titers of the positive patients in each stage were also not different from those in each other stage. These results indicated that HD antigenic expression on cancerous tissue is not dependent on the cancerous malignancy. The HD antibody level was elevated after surgical removal of cancerous tissues in 5 of 6 patients examined, indicating that tumor growth absorbed the serum antibody. Serum antibody against 4-O-acetyl-HD3 was detected independently of HD3 antibody in some cases however, in most cases, correlation between the two antibody titers was observed.
  • Anti-murine IL-6 receptor antibody inhibits IL-6 effects in vivo, H. Suzuki, K. Yasukawa, T. Saito, M. Anzai, R. Goitsuka, A. Hasegawa, Y. Ohsugi, T. Taga, T. Kishimoto, Immunology Letters, Immunology Letters, 30(1), 17 - 22, 1991 , Refereed
    Summary:Thrombopoiesis, as well as antibody production, is one of the major events in which interleukin-6 (IL-6) has been reported to be involved. Polyclonal antimurine IL-6 receptor antibody was prepared to examine the effect of the antibody on these events in IL-6-treated mice. Administration of the anti-mIL-6R antibody inhibited the IL-6-induced increase in the number of platelets. Enhancement of the serum level of DNP-specific antibody by intraperitoneal injection of IL-6 was inhibited completely with simultaneous administration of the anti-mIL-6R antibody. The level of DNP-specific antibody was decreased, even below the basal value, by the higher dose of anti-mIL-6R antibody, indicating its effect also on endogenous IL-6. This work provides evidence that anti-IL-6R antibody inhibits IL-6 function in vivo, and provides an animal model of the therapeutic use of anti-IL-6R antibody for IL-6-related disease. © 1991.

Conference Activities & Talks

  • Nuclear dynamics in chromosome territories and gene loci under in vitro differentiation of mouse embryonic stem cells, 44th Annual Meeting for the Japanese Society of Developmental Biologists,   2011 05 , 44th Annual Meeting for the Japanese Society of Developmental Biologists
  • Non-B DNA segment promotes high and stable transcription activity of the transgene in mouse embryonic stem cells, 44th Annual Meeting for the Japanese Society of Developmental Biologists,   2011 05 , 44th Annual Meeting for the Japanese Society of Developmental Biologists
  • Expression of the components of chromatin remodeling factor SWR1 complex in mouse somatic nuclear transfer eggs,   2009 12
  • Dynamics of Arp family proteins and components of chromatin remodeling complexes in in vitro differentiation of mouse embryonic stem cells,   2009 12
  • FGF4 modulates the development of mouse somatic nuclear transfer embryos,   2009 12
  • Reconstitution of seminiferous tubules in xenoectopic transplantation with bovine testicular cells, 42nd Annual Meeting for the Japanese Society of Developmental Biologists,   2009 05 , 42nd Annual Meeting for the Japanese Society of Developmental Biologists
  • Expression of Bcrp1 mRNA isoforms during in vitro differentiation of mouse embryonic stem cells, 41st Annual Meeting for the Japanese Society of Developmental Biologists,   2008 05 , 41st Annual Meeting for the Japanese Society of Developmental Biologists
  • Expression of transcription factor Cdx2 and Oct3/4 in mouse somatic cell nuclear transfer embryos.,   2007 05
  • 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
  • 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
  • Expression of maternal gene promoter driven expression vectors in mouse oocytes and fertilized eggs, 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
  • 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

Misc

  • 初期胚特異的レトロトランスポゾンMERVLの活性化機構の解析, 奥野智美, 樋口智香, 神谷拓磨, 山本真理, 越智浩介, 西野亜理紗, 井橋俊哉, 辻本佳加理, 松橋珠子, 安齋政幸, 黒坂哲, 三谷匡, 山縣一夫, 細井美彦, 松本和也, 宮本圭, Journal of Reproduction and Development, 64, Suppl Japanese Issue, j125, 70-P-70,   2018 09 05 , https://jglobal.jst.go.jp/detail?JGLOBAL_ID=201802285393389434
    Summary:<p>【目的】哺乳類ゲノムの約半分は,トランスポゾン由来の配列で構成されている。特に,Murine Endogenous RetroVirus-L(MERVL)と呼ばれるレトロトランスポゾンは,マウス2細胞期胚にかけてその発現が一過的に上昇する独特な発現様式を示し,2細胞期胚特異的に発現する遺伝子群の転写活性を制御している。また,マウスES細胞においてもMERVLを発現する細胞群が存在する。しかし,MERVLの活性化機構の全容は未だ明らかにされていない。そこで本研究では,マウスES細胞を用いて,MERVLの遺伝子発現調節領域(LTR領域)の活性化が誘導される条件を検討した。【方法】マウスES細胞E14Tg2a株を用い,ヒストン脱アセチル化酵素阻害剤であるTrichostatin A(TSA)及びクロマチンのメチル化状態を変化させるVitamin C(VC)を培地に添加した。まず,10 nM TSA添加区,10 ng/µl VC添加区,10 nM TSA及び10 ng/µl VC添加区において,RT-qPCRを用いて2細胞期胚で強発現する遺伝子の転写量を比較した。さらに,2C::td Tomatoベクターをゲノム上に組み込み,MERVL-LTR領域の活性化を可視化できるES細胞を用いて,各培養条件下におけるMERVL-LTR領域活性化細胞数の割合を調べた。【結果】7日間TSAを添加した培養条件下において,2細胞期胚特異的遺伝子群の転写量が有意に上昇していた。また,TSA処理継続時間に比例してMERVL-LTR領域活性化細胞数の割合が約5%まで増加することがわかった。以上の結果より,ES細胞においてMERVL-LTR領域を活性化する培養条件を示した。また,TSA処理によるMERVL活性化促進の結果より,ゲノムワイドなクロマチン弛緩がMERVL-LTR領域の活性化につながる可能性が示唆された。現在,クロマチン脱凝縮を誘導する因子の過剰発現がMERVL活性化を促すか検討を進めている。</p>
  • レトロトランスポゾンMERVLの活性化機構の解析, 奥野智美, 山口壮輝, 樋口智香, 神谷拓磨, 山本真理, 越智浩介, 西野亜理紗, 井橋俊哉, 辻本佳加理, 坂本裕子, 松橋珠子, 安齋政幸, 黒坂哲, 三谷匡, 山縣一夫, 細井美彦, 松本和也, 宮本圭, 日本分子生物学会年会プログラム・要旨集(Web), 41st, ROMBUNNO.2P‐0413 (WEB ONLY),   2018 , https://jglobal.jst.go.jp/detail?JGLOBAL_ID=201802273880748976
  • Efficient nuclear reprogramming of somatic cells towards totipotency is supported by synergistic effects of small molecules., K. Miyamoto, Y. Tajima, K. Yoshida, M. Oikawa, R. Azuma, M. Mori, Y. Imasato, G.E. Allen, T. Tsujikawa, T. Tsukaguchi, C.R. Bradshaw, J. Jullien, K. Yamagata, K. Matsumoto, M. Anzai, H. Imai, J.B. Gurdon, M. Yamada, the 50th annual meeting of Japan Society of Developmental Biologists,   2017 05 , Refereed, 招待有り
  • 受精後のユビキチン・プロテアソーム系による母性タンパク質の分解はマウス初期胚発生に重要である, 樋口智香, 守田昂太郎, 山口壮輝, 松橋珠子, 永井宏平, 安齋政幸, 安齋政幸, 山縣一夫, 細井美彦, 宮本圭, 松本和也, 日本卵子学会誌, 2, 1, S48, S48,   2017 04 01 , http://jglobal.jst.go.jp/public/201702240011137048
  • マウス初期胚発生過程におけるH3R2me2sの役割, 守田昂太郎, 樋口智香, 山口壮輝, 松橋珠子, 松橋珠子, 永井宏平, 安齋政幸, 安齋政幸, 加藤博巳, 加藤博巳, 山縣一夫, 細井美彦, 宮本圭, 松本和也, 日本卵子学会誌, 2, 1, S47, S47,   2017 04 01 , http://jglobal.jst.go.jp/public/201702274754974238
  • The effect of Coenzyme Q10 on in vitro fertilization and subsequent preimplantation development of post-ovulatory aged oocytes in mice, UCHIBORI Sho, HIGUCHI Chika, MORITA Kohtaro, TSUKAGUCHI Tomomasa, ANZAI Masayuki, YAMAGATA Kazuo, HOSOI Yoshihiko, MIYAMOTO Kei, MATSUMOTO Kazuya, The Journal of Reproduction and Development Supplement, 108, 0, P, 10-P-10,   2015 , http://ci.nii.ac.jp/naid/130005492071
  • Availability of genetic resource using somatic cell nuclei derived from muscle tissues by expression of phosphorylated H2A.X, AZUMA Rika, MIYASHITA Minoru, NAGAI Kouhei, NAKAGAWA Takao, KAJIMOTO Mizuki, INOUE Tatsuya, HOSOI Yoshihiko, ANZAI Masayuki, The Journal of Reproduction and Development Supplement, 108, 0, P, 81-P-81,   2015 , http://ci.nii.ac.jp/naid/130005492100
  • Involvement of Peroxiredoxin (Prdx) in reducing hydrogen peroxide in pronuclei of mouse zygotes., MORITA Kohtaro, TOKORO Mikiko, HIGUCHI Chika, UCHIBORI Sho, TSUKAGUCHI Tomomasa, NAGAI Kohei, ANZAI Masayuki, YAMAGATA Kazuo, HOSOI Yoshihiko, MIYAMOTO Kei, MATSUMOTO Kazuya, The Journal of Reproduction and Development Supplement, 108, 0, P, 15-P-15,   2015 , http://ci.nii.ac.jp/naid/130005492029
  • A transient inhibition of proteasome pathway entails a delay in the onset of ZGA and impairs full term development of mice, HIGUCHI Chika, SHIMIZU Natsumi, MORITA Kohtaro, UCHIBORI Sho, TSUKAGUCHI Tomomasa, NAGAI Kohei, ANZAI Masayuki, YAMAGATA Kazuo, HOSOI Yoshihiko, MIYAMOTO Kei, MATSUMOTO Kazuya, The Journal of Reproduction and Development Supplement, 108, 0, P, 12-P-12,   2015 , http://ci.nii.ac.jp/naid/130005492035
  • マウスGV期由来卵子を用いた体外成熟後の卵子母性制御機構に関する基礎検討, SAKITA MEGUMI, KAMEI MIKU, NAKAGAWA TAKAO, NISHIMURA MANAMI, NAKAIE MASATAKA, KOBAYASHI SHINTARO, HIGASHI RIKA, MITANI TASUKU, KATO HIROMI, KISHI MASAO, HOSOI YOSHIHIKO, ANZAI MASAYUKI, 日本実験動物学会総会講演要旨集, 61st, 245,   2014 05 01 , http://jglobal.jst.go.jp/detail.php?from=API&JGLOBAL_ID=201402283057517607
  • 体外成熟培地へのアミノ酸誘導体添加がマウス未成熟卵子由来初期胚の発生に与える効果, KAMEI MIKU, SAKITA MEGUMI, NAKAGAWA TAKAO, NAKAIE MASATAKA, NISHIMURA MANAMI, HIGASHI RIKA, KOBAYASHI SHINTARO, MITANI TASUKU, KATO HIROMI, KISHI MASAO, HOSOI YOSHIHIKO, ANZAI MASAYUKI, 日本実験動物学会総会講演要旨集, 61st, 246,   2014 05 01 , http://jglobal.jst.go.jp/detail.php?from=API&JGLOBAL_ID=201402287287945670
  • Reconstitution of seminiferous tubules by xenoectopic transplantation of juvenile bovine testicular cells into the subcutis of immunodeficient mice, 喜多 章太, 森木 甲子郎, 谷口 俊仁, 安齋 政幸, 加藤 博己, 佐伯 和弘, 細井 美彦, 三谷 匡, Memoirs of Institute of Advanced Technology, Kinki University, 18, 29, 38,   2013 03 , http://ci.nii.ac.jp/naid/120005260089
    Summary:[要旨] 1994年、精原幹細胞の精細管移植によりドナー由来の精子を作製できる移植技術が開発された。さらに、2007年には幼若ブタ精巣細胞を免疫不全マウスの皮下へ移植することにより、精子形成の誘導に成功したことが報告された。そこで本研究では、絶滅危惧動物などでの精子形成分化誘導モデルの確立を目的に、ウシ精巣細胞を用いた移植実験を行った。マウス新生子(生後7日以内)、ウシ新生子(生後7 日以内)およびウシ幼若(3~5ヶ月齢)精巣細胞をマトリゲルマトリックスと混合し、免疫不全マウスの皮下に移植することで、精巣様構造の再構築と精子形成の誘導を試みた。移植後、1週~ 44週に組織学的解析を行った。さらに、生殖細胞の生存・増殖・分化について、抗VASA 抗体を用いた免疫組織化学的解析を行った。マウス新生子精巣細胞の移植では、移植後10週以降に再構築精細管および減数分裂に至る精子細胞がみられ、精子形成誘導を可能にする機能的な精細管の構築が示された。一方、新生子ならびに幼若ウシ精巣細胞の移植では、移植後10 週から管構造の再建がみられたものの、免疫組織化学的解析において再構築された精細管様構造の基底膜上に配置される生殖細胞を認めることはできなかった。以上の結果から、異種異所移植法によりウシにおいても精細管様構造が再構築されたことから, 今後移植法の改良によりウシ精子形成の誘導が可能となることが期待される。 [Abstract] Male germ cell transplantation induces donor cell-derived spermatogenesis in the testis of recipient mice. However, this technique is still available only to mice and rats. Recently, xenoectopic transplantation of testicular cells into the subcutis of immunodeficient mice has been developed as a novel technology to rebuild seminiferous tubules and to produce functional sperm. In order to develop an alternative technology for transgenesis and induction of spermatogenesis in domestic animals, we examinedxenoectopic transplantation of bovine testicular cells. Testicular cells from neonatal(within 7 days)and juvenile(3-5 months)bovine and neonatal(within 7 days)mouse testes were prepared at the concentration of 1-5 x 10^7/ml(mouse)or 1 x 10^8/ml(bovine)and then mixed with equal volume of BD Matrigel^<TM> Basement Membrane Matrix(Matrigel). The testicular cell/Matrigel suspensions were grafted into the subcutis of castrated immunodeficient male mice(BALB/c Slc-nu/nu). Morphogenesis of the grafts was examined between 1 and 44 weeks after transplantation. To examine the growth and differentiation of donor germ cells, the expression of VASA, which is expressed from gonocytes to early meiotic stages in mammalian male germ cells, was examined by immunohistochemistry. Grafts derived from mouse testicular cells regenerated complete functional seminiferous tubules 20 weeks after transplantation in which haploid spermatids were generated in the lumen. Initiation of reconstruction of seminiferous tubules from bovine testicular cells was also observed 10 weeks after transplantation. However, immunohistochemical analysis demonstrated that bovine germ cells failed to colonize along the basement membrane of reconstructed seminiferous tubules and to proliferate thereafter. Although further investigation for suitable conditions of bovine germ cell colonization and survival in the reconstructed seminiferous tubules is needed, xenoectopic transplantation would make possible to induce bovine spermatogenesis and be usefull for animal sciences, conservation of wild animals and male infertility.
  • マウス初期胚の第一分裂におけるRhophilin‐2の関与, MORITA KOTARO, HATANAKA YUKI, SHIMIZU NATSUMI, NISHIKAWA SATOSHI, NISHIHARA TAKASHI, KATO RIE, TAKEMOTO ATSUSHI, HIGUCHI CHIKA, AMANO TOMOKO, ANZAI MASAYUKI, KISHIGAMI SATOSHI, SAEKI KAZUHIRO, HOSOI YOSHIHIKO, MATSUMOTO KAZUYA, 日本はい移植学雑誌, 35, 1, 31, 31,   2013 01 21 , http://jglobal.jst.go.jp/detail.php?from=API&JGLOBAL_ID=201302232038187000
  • 近交系マウス由来未成熟卵子を用いた生殖補助技術により作出した各卵子母性制御機構に関する検討, ANZAI MASAYUKI, NISHIMURA MANAMI, AZUMA RIKA, NAKAIE MASATAKA, KAMEI MIKU, SAKITA MEGUMI, KATO HIROMI, MITANI TASUKU, HOSOI YOSHIHIKO, 日本分子生物学会年会プログラム・要旨集(Web), 36th, 1P-0669 (WEB ONLY),   2013 , http://jglobal.jst.go.jp/detail.php?from=API&JGLOBAL_ID=201402206788995589
  • マウス初期胚におけるユビキチン・プロテアソーム経路の転写制御への関与, SHIMIZU NATSUMI, HATANAKA YUKI, HIGUCHI CHIKA, SHIN SEUNG-WOOK, NISHIKAWA SATOSHI, NISHIHARA TAKUJI, KATO RIE, TAKEMOTO ATSUSHI, MORITA KOTARO, NAGAI KOHEI, AMANO TOMOKO, KISHIGAMI SATOSHI, ANZAI MASAYUKI, HOSOI YOSHIHIKO, MATSUMOTO KAZUYA, 日本分子生物学会年会プログラム・要旨集(Web), 35th, 3P-0488 (WEB ONLY),   2012 , http://jglobal.jst.go.jp/detail.php?from=API&JGLOBAL_ID=201302205140673647
  • A role of ubiquitin-proteasome system in early mouse embryos, SHIMIZU Natsumi, HIGUCHI Chika, HATANAKA Yuki, SHIN Seung-Wook, NISHIKAWA Satoshi, NISHIHARA Takuji, KATO Rie, TAKEMOTO Atsushi, MORITA Kotaro, AMANO Tomoko, KISHIGAMI Satoshi, ANZAI Masayuki, SAEKI Kazuhiro, HOSOI Yoshihiko, MATSUMOTO Kazuya, The Journal of Reproduction and Development Supplement, 105, 0,   2012 , http://ci.nii.ac.jp/naid/130005457604
  • GSE, predicted histone chaperone, involves in active DNA demethylation in early mouse embryos, HATANAKA Yuki, MORITA Kotarou, SHIMIZU Natsumi, NISHIKAWA Satoshi, NISHIHARA Takuji, KATOU Rie, TAKEMOTO Atsushi, HIGUCHI Chika, AMANO Tomoko, ANZAI Masayuki, KISHIGAMI Satoshi, SAEKI Kazuhiro, HOSOI Yoshihiko, MATSUMOTO Kazuya, The Journal of Reproduction and Development Supplement, 105, 0,   2012 , http://ci.nii.ac.jp/naid/130005457594
  • Rhophilin-2 associating with GABARAP is involved during the first cleavage of early mouse embryo under the control of RhoB., MORITA Kotarou, HATANAKA Yuki, SHIMIZU Natsumi, NISHIKAWA Satoshi, NISHIHARA Takuji, KATOU Rie, TAKEMOTO Atsushi, HIGUCHI Chika, AMANO Tomoko, ANZAI Masayuki, KISHIGAMI Satoshi, SAEKI Kazuhiro, HOSOI Yoshihiko, MATSUMOTO Kazuya, The Journal of Reproduction and Development Supplement, 105, 0,   2012 , http://ci.nii.ac.jp/naid/130005457539
  • 卵子活性化に伴うチューブリンのアセチル化の解析, 松原 圭吾, Lee Ah Reum, 鎌田 悠, 孫谷 匡輝, 奥山 紀之, 三谷 匡, 加藤 博己, 安齋 政幸, 佐伯 和弘, 松本 和也, 入谷 明, 岸上 哲士, 細井 美彦, 日本生化学会大会・日本分子生物学会年会合同大会講演要旨集, 83回・33回, 1P, 0889,   2010 12
  • 卵子および一細胞期胚におけるタンパク質のアセチル化に関する解析, MATSUBARA KEIGO, LEE AH REUM, KAMATA YU, MAGOTANI MASATERU, MITANI TASUKU, KATO HIROMI, ANZAI MASAYUKI, SAEKI KAZUHIRO, MATSUMOTO KAZUYA, IRITANI AKIRA, KISHIGAMI SATOSHI, HOSOI YOSHIHIKO, J Reprod Dev, 56, Suppl Japanese Issue, J64,   2010 08 20 , 10.14882/jrds.103.0.12.0, http://jglobal.jst.go.jp/detail.php?from=API&JGLOBAL_ID=201002250746875672
  • マウス初期胚における発生制御タンパク質の解析, 野老 美紀子, 川澄 みゆり, 永井 宏平, 池上 春香, 申 承旭, 西川 慧, 畑中 勇輝, 西原 卓志, 天野 朋子, 三谷 匡, 加藤 博巳, 安齋 政幸, 岸上 哲士, 佐伯 和弘, 細井 美彦, 入谷 明, 松本 和也, The Journal of Reproduction and Development, 56, Suppl., j120, j120,   2010 08 , 10.14882/jrds.103.0.124.0
  • Influence of insemination concentration when in vitro fertilization after zona drilled by laser with C57BL/6 in vitro maturation oocytes, NISHIMURA Manami, NISHIYAMA Yui, YANAGI Miho, KAWABE Toshiaki, MITANI Tasuku, HOSOI Yoshihiko, ANZAI Masayuki, 27, 2,   2010 04 , http://ci.nii.ac.jp/naid/10026350802
  • マウス胚性幹細胞におけるABCトランスポーター・Bcrp1mRNAアイソフォームAの転写制御領域の解析, HIRANO DAIKI, KAWAMURA HIROKO, NISHIMURA YUI, SAEKI KEITA, ANZAI MASAYUKI, KATO HIROMI, TAGUCHI YOSHITOMO, HOSOI YOSHIHIKO, IRITANI AKIRA, MITANI TASUKU, 生化学, ROMBUNNO.4P-0843,   2010 , http://jglobal.jst.go.jp/detail.php?from=API&JGLOBAL_ID=201002274122783214
  • マウス初期胚におけるDD2‐2遺伝子は2OSプロテアソームの形成に関与している, SHIN SHOKYOKU, TOKORO MIKIKO, NISHIKAWA KEI, RI KOKIN, HATANAKA YUKI, AMANO TOMOKO, MITANI TASUKU, KATO HIROMI, ANZAI MASAYUKI, KISHIGAMI SATOSHI, SAEKI KAZUHIRO, HOSOI YOSHIHIKO, IRITANI AKIRA, MATSUMOTO KAZUYA, 日本畜産学会大会講演要旨, 111th, 78,   2009 09 28 , http://jglobal.jst.go.jp/detail.php?from=API&JGLOBAL_ID=200902223084547469
  • マウスの着床前胚におけるユビキチンプロテアソーム経路による母体蛋白質変性の解析(Analysis of degraded maternal proteins by ubiquitin-proteasome pathway in mouse preimplantation embryo), 李 香欣, 申 承旭, 野老 美紀子, 西川 慧, 畑中 勇輝, 天野 朋子, 岸上 哲士, 安齋 政幸, 三谷 匡, 加藤 博己, 佐伯 和弘, 細井 美彦, 入谷 明, 松本 和也, 日本畜産学会大会講演要旨集, 111回, 78, 78,   2009 09
  • マウス初期胚におけるDD2-2遺伝子は20Sプロテアソームの形成に関与している, 申 承旭, 野老 美紀子, 西川 慧, 李 香欣, 畑中 勇輝, 天野 朋子, 三谷 匡, 加藤 博己, 安齋 政幸, 岸上 哲士, 佐伯 和弘, 細井 美彦, 入谷 明, 松本 和也, 日本畜産学会大会講演要旨集, 111回, 78, 78,   2009 09
  • プロテオミクスによるマウス着床前期胚の発生関連タンパク質の解析, 野老 美紀子, 川澄 みゆり, 永井 宏平, 池上 春香, 申 承旭, 西川 慧, 李 香欣, 畑中 勇輝, 天野 朋子, 三谷 匡, 加藤 博己, 安齋 政幸, 岸上 哲士, 佐伯 和弘, 細井 美彦, 入谷 明, 松本 和也, 日本畜産学会大会講演要旨集, 111回, 78, 78,   2009 09
  • FGF4がマウス体細胞核移植胚の発生に与える効果, MORITA MASAHIRO, ANZAI MASAYUKI, NISHIYAMA YUI, KATO HIROMI, KISHIGAMI SATOSHI, HOSOI YOSHIHIKO, MITANI TAKUMI, IRIYA AKIRA, J Reprod Dev, 55, Supplement, J84,   2009 08 25 , 10.14882/jrds.102.0.134.0, http://jglobal.jst.go.jp/detail.php?from=API&JGLOBAL_ID=200902229379782638
  • マウスES細胞の未分化維持機構においてABCトランスポーターBcrp1が与える影響, KAWAMURA HIROKO, KAWAI TOMOKO, TAGUCHI YOSHITOMO, ANZAI MASAYUKI, KATO HIROKI, HOSOI YOSHIHIKO, MITANI TADASHI, IRIYA AKIRA, J Reprod Dev, 55, Supplement, J79,   2009 08 25 , 10.14882/jrds.102.0.123.0, http://jglobal.jst.go.jp/detail.php?from=API&JGLOBAL_ID=200902291185683946
  • マウス初期胚におけるDD2-2遺伝子は20Sプロテアソームの形成に関与している, 申 承旭, 野老 美紀子, 西川 慧, 李 香欣, 畑中 勇輝, 天野 朋子, 三谷 匡, 加藤 博己, 安齋 政幸, 岸上 哲士, 佐伯 和弘, 細井 善彦, 入谷 明, 松本 和也, The Journal of Reproduction and Development, 55, Suppl., j74, j74,   2009 08
  • Sr2+が誘導するマウス卵子活性化へのカルシウムキレート剤の利用, 辻本 賀子, 岸上 哲士, 竹原 俊幸, 天野 朋子, 安齋 政幸, 加藤 博巳, 三谷 匡, 松本 和也, 佐伯 和弘, 入谷 明, 細井 美彦, The Journal of Reproduction and Development, 55, Suppl., j84, j84,   2009 08 , 10.14882/jrds.102.0.133.0
  • 若齢ウシ精巣細胞を用いたヌードマウス皮下への異種異所移植による精子形成誘導の試み, 森木 甲子郎, 喜多 章多, 藤本 佑希, 谷口 俊仁, 安齋 政幸, 加藤 博己, 松本 和也, 佐伯 和弘, 細井 美彦, 三谷 匡, 入谷 明, The Journal of Reproduction and Development, 55, Suppl., j96, j96,   2009 08 , 10.14882/jrds.102.0.220.0
  • プロテオミクスを用いたマウス初期胚における発生関連タンパク質の解析, 野老 美紀子, 川澄 みゆり, 永井 宏平, 池上 春香, 申 承旭, 西川 慧, 李 香欣, 畑中 勇輝, 天野 朋子, 三谷 匡, 加藤 博己, 安齋 政幸, 岸上 哲士, 佐伯 和弘, 細井 美彦, 入谷 明, 松本 和也, The Journal of Reproduction and Development, 55, Suppl., j142, j142,   2009 08 , 10.14882/jrds.102.0.1075.0
  • マウス初期胚でのユビキチン-プロテアソーム系による母性タンパク質の分解に関する研究, 李 香欣, 申 承旭, 野老 美紀子, 西川 慧, 畑中 勇輝, 天野 朋子, 岸上 哲士, 安齋 政幸, 三谷 匡, 加藤 博己, 佐伯 和弘, 細井 美彦, 入谷 明, 松本 和也, The Journal of Reproduction and Development, 55, Suppl., j143, j143,   2009 08
  • Birth of mice from vitrified/warmed 2-cell embryos transported at a cold temperature, NAKAGATA Naomi, TAKEO Toru, KANEKO Takehito, HARUGUCHI Yukie, FUKUMOTO Kiyoko, MACHIDA Hiromi, KOGA Mika, NAKAGAWA Yoshiko, TAKESHITA Yumi, MATSUGUMA Toyokazu, TSUCHIYAMA Shuuji, SHIMIZU Norihiko, HASEGAWA Takanori, GOTO Nobuhito, MIYACHI Hitoshi, ANZAI Masayuki, NAKATSUKASA Ena, NOMARU Kouji, 26, 2, S48,   2009 04 01 , http://ci.nii.ac.jp/naid/10024964983
  • 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, 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。
  • マウスES細胞の分化過程における未分化維持機構においてABCトランスポーターBcrp1の影響, KAWAMURA HIROKO, KAWAI TOMOKO, MORIKI KOSHIRO, TAGUCHI YOSHITOMO, ANZAI MASAYUKI, KATO HIROMI, HOSOI YOSHIHIKO, IRITANI AKIRA, MITANI TASUKU, 日本分子生物学会年会講演要旨集, 32nd, Vol.1, 191,   2009 , http://jglobal.jst.go.jp/detail.php?from=API&JGLOBAL_ID=201002212646226678
  • マウスES細胞の分化過程におけるArpファミリーならびにクロマチンリモデリング複合体構成因子の動態, MORIKI KOSHIRO, NISHIYAMA YUI, KAWAMURA HIROKO, ANZAI MASAYUKI, KATO HIROMI, HOSOI YOSHIHIKO, HARATA MASAHIKO, IRITANI AKIRA, MITANI TASUKU, 日本分子生物学会年会講演要旨集, 32nd, Vol.1, 190,   2009 , http://jglobal.jst.go.jp/detail.php?from=API&JGLOBAL_ID=201002222527720821
  • FGF4はマウス体細胞核移植胚の初期発生を促進する, MORITA MASAHIRO, ANZAI MASAYUKI, NISHIYAMA YUI, KATO HIROMI, KISHIGAMI SATOSHI, SAEKI KAZUHIRO, HOSOI YOSHIHIKO, IRITANI AKIRA, MITANI TASUKU, 日本分子生物学会年会講演要旨集, 32nd, Vol.1, 179,   2009 , http://jglobal.jst.go.jp/detail.php?from=API&JGLOBAL_ID=201002238117619833
  • トランスジェニックマウスを用いた母性遺伝子プロモーター解析, 畑中 勇輝, 中野 彰大, 常本 和伸, 安齋 政幸, 佐藤 学, 野老 美紀子, 申 承旭, 渡辺 達也, 清水 なつみ, 天野 朋子, 三谷 匡, 加藤 博己, 岸上 哲士, 細井 美彦, 佐伯 和弘, 入谷 明, 松本 和也, The Journal of Reproduction and Development, 54, Suppl., j75, j75,   2008 08 , 10.14882/jrds.101.0.213.0
  • マウス初期胚でDD2-2遺伝子は20Sプロテアソームの形成に関与している, 申 承旭, 野老 美紀子, 佐藤 学, 西川 慧, 天野 朋子, 岸上 哲士, 安齋 政幸, 三谷 匡, 加藤 博己, 佐伯 和弘, 細井 美彦, 入谷 明, 松本 和也, The Journal of Reproduction and Development, 54, Suppl., j76, j76,   2008 08
  • マウス着床前期胚における大規模プロテオーム解析, 野老 美紀子, 川澄 みゆり, 永井 宏平, 池上 春香, 佐藤 学, 申 承旭, 西川 慧, 清水 なつみ, 天野 朋子, 三谷 匡, 加藤 博己, 安齋 政幸, 岸上 哲士, 佐伯 和弘, 細井 美彦, 入谷 明, 松本 和也, The Journal of Reproduction and Development, 54, Suppl., j95, j95,   2008 08 , 10.14882/jrds.101.0.507.0
  • Promoter activity of three maternal-effect genes in transgenic mice, NAKANO Akihiro, TSUNEMOTO Kazunobu, ANZAI Masayuki, SATO Manabu, TOKORO Mikiko, SHIN Seungwook, HATANAKA Yuki, AMANO Tomoko, MITANI Tasuku, KATO Hiromi, HOSOI Yoshihiko, SAEKI Kazuhiro, IRITANI Akira, MATSUMOTO Kazuya, 25, 2,   2008 04 01 , http://ci.nii.ac.jp/naid/10021218252
  • Expression of Bcrp1 mRNA isoforms of mouse embryonic stem cells, KAWAMURA HIROKO, AMIMOTO NAOKI, TAGUCHI YOSHITOMO, ANZAI MASAYUKI, KATO HIROMI, IRITANI AKIRA, MITANI TASUKU, 近畿大学先端技術総合研究所紀要, 13, 29, 40,   2008 03 31 , http://jglobal.jst.go.jp/detail.php?from=API&JGLOBAL_ID=200902223230443387
  • Induction of three germ layer cells from parthenogenetic or androgenetic embryo origin embryonic stem cells in vitro, 小野寺 勇太, 寺村 岳士, 竹原 俊幸, 村上 秀樹, 小澤 まどか, 武内 大輝, 安齋 政幸, 加藤 博己, 三谷 匡, 松本 和也, 佐伯 和弘, 入谷 明, 佐川 典正, 細井 美彦, Memoirs of Institute of Advanced Technology, Kinki University, 13, 9, 19,   2008 03 , http://ci.nii.ac.jp/naid/120005736395
    Summary:胚性幹細胞(Embryonic Stem Cell ; ES 細胞)は自己複製能と様々な細胞へと分化することの出来る分化多能性を有している細胞である。近年、このES 細胞を用いた再生修復医療の可能性が注目されている。しかし、受精卵からES 細胞を作出するため、移植後に免疫拒絶反応を示すことが示唆されており、ES 細胞を用いた再生修復医療の1 つの課題となっている。そこで、片親性の雌性単為発生胚と雄性発生胚からES 細胞を樹立し、片親性ES 細胞の分化多能性を観察した。
  • Expression of stem and germ cell markers in juvenile bovine testis and its primary culture, 森木 甲子郎, 田津原 陽平, 谷口 俊二, 安齋 政幸, 加藤 博己, 佐伯 和弘, 入谷 明, 三谷 匡, Memoirs of Institute of Advanced Technology, Kinki University, 13, 41, 50,   2008 03 , http://ci.nii.ac.jp/naid/120005736398
    Summary:近年、マウスやラットにおいて精子形成を通して次世代の個体構築を担う唯一の幹細胞である精原幹細胞を体外で培養し、株化した生殖幹細胞(Germline Stem Cell;GS 細胞)が樹立された。しかし、ウシなどの食資源動物ではES 細胞やEG 細胞はいまだ樹立されておらず、高度な遺伝子改変を行う上で、代替えとなる幹細胞の樹立が望まれる。ウシ雄性生殖細胞の同定には、ヘマトキシリン・エオシン染色やレクチンの一種であるDBA やc-kit を用いた組織学的解析やフローサイトメトリー(FCM)による解析が主に行われてきた。これにより、ウシ精巣内のゴノサイトや精原幹細胞やA 型精原細胞が、同定されてきた。そこで本研究では、ウシにおけるゴノサイトおよび精原幹細胞の同定を目的として、RT-PCR ならびにIn situ hybridization 法により、未分化マーカーとされているOct3/4 のmRNA の検出を試みた。さらに、ゴノサイトで発現しているVASA タンパク質に注目し、月齢の異なるウシ精巣内、ならびに初代培養したin vitro でのゴノサイトの局在を免疫組織化学染色により同定した。その結果、ウシOct3/4 は、RT-PCR において分化細胞からも検出され、さらにIn situ hybridization においても、精細管内で特異的な発現は認められなかった。一方、VASA タンパク質は若齢ウシ精巣においてマウスと同様の発現パターンを示し、またウシ精巣細胞の初代培養においてもVASA 陽性細胞の増殖が認められた。これらの結果から、ウシ若齢精巣において、VASA はゴノサイトや精原幹細胞で発現し、体外培養法の開発にも有効であることが示された。
  • In vitroにおける単為発生/雄性発生胚由来ES細胞からの三胚葉由来機能細胞の獲得, ONODERA YUTA, TERAMURA TAKESHI, TAKEHARA TOSHIYUKI, FUKUNAGA NAOTO, ITO SHUNSUKE, MURAKAMI HIDEKI, ANZAI MASAYUKI, KATO HIROMI, MITANI TADASHI, MATSUMOTO KAZUYA, SAEKI KAZUHIRO, KISHIGAMI TETSUJI, IRITANI AKIRA, SAGAWA NORIMASA, HOSOI YOSHIHIKO, 再生医療, 7, 242,   2008 02 22 , http://jglobal.jst.go.jp/detail.php?from=API&JGLOBAL_ID=200902221602095710
  • 雌性単為発生胚と雄性単為発生胚由来ES細胞から誘導したインスリン産生細胞の機能性の検討, TAKEUCHI HIROKI, TERAMURA TAKESHI, KISHIGAMI TETSUJI, ANZAI MASAYUKI, KATO HIROMI, MITANI TADASHI, MATSUMOTO KAZUYA, SAEKI KAZUHIRO, SAGAWA NORIMASA, HOSOI YOSHIHIKO, 再生医療, 7, 241,   2008 02 22 , http://jglobal.jst.go.jp/detail.php?from=API&JGLOBAL_ID=200902242585690223
  • マウス着床前期胚における大規模プロテオーム解析, TOKORO MIKIKO, KAWASUMI MIYURI, NAGAI KOHEI, IKEGAMI HARUKA, SATO MANABU, SHIN SEUNG-WOOK, NISHIKAWA SATOSHI, AMANO TOMOKO, MITANI TASUKU, KATO HIROMI, ANZAI MASAYUKI, KISHIGAMI SATOSHI, SAEKI KAZUHIRO, HOSOI YOSHIHIKO, IRITANI AKIRA, MATSUMOTO KAZUYA, 生化学, 4P-0894,   2008 , http://jglobal.jst.go.jp/detail.php?from=API&JGLOBAL_ID=200902223409224866
  • 若齢ウシ雄性生殖細胞の同定ならびにヌードマウス皮下移植による精子形成誘導の試み, MORIKI KOSHIRO, KITA SHOTA, TANIGUCHI SHUNJI, ANZAI MASAYUKI, KATO HIROMI, SAEKI KAZUHIRO, HOSOI YOSHIHIKO, IRITANI AKIRA, MITANI TASUKU, 生化学, 3P-0886,   2008 , http://jglobal.jst.go.jp/detail.php?from=API&JGLOBAL_ID=200902227490032416
  • トリコスタチンA処理によるマウス体細胞核移植胚の着床初期における胚発生に関わる遺伝子発現の解析, MORITA MASAHIRO, NISHIWAKI MEGUMI, ANZAI MASAYUKI, NISHIYAMA YUI, KATO HIROMI, KISHIGAMI SATOSHI, SAEKI KAZUHIRO, HOSOI YOSHIHIKO, IRITANI AKIRA, MITANI TASUKU, 生化学, 4P-0895,   2008 , http://jglobal.jst.go.jp/detail.php?from=API&JGLOBAL_ID=200902237521150191
  • マウス初期胚で性腺特異的遺伝子DD2‐2が20Sプロテアソームの形成に及ぼす影響, SHIN SEUNG-WOOK, TOKORO MIKIKO, SATO MANABU, NISHIKAWA SATOSHI, AMANO TOMOKO, KISHIGAMI SATOSHI, ANZAI MASAYUKI, MITANI TASUKU, KATO HIROMI, SAEKI KAZUHIRO, HOSOI YOSHIHIKO, IRITANI AKIRA, MATSUMOTO KAZUYA, 生化学, 4P-0892,   2008 , http://jglobal.jst.go.jp/detail.php?from=API&JGLOBAL_ID=200902285509934742
  • マウス排卵卵子における網羅的タンパク質発現(プロテオーム)解析, 野老 美紀子, 川澄 みゆり, 永井 宏平, 池上 春香, 申 承旭, 松岡 俊樹, 佐藤 学, 天野 朋子, 三谷 匡, 加藤 博巳, 安齋 政幸, 岸上 哲士, 佐伯 和弘, 細井 美彦, 入谷 明, 松本 和也, The Journal of Reproduction and Development, 53, Suppl., j137, j137,   2007 09 , 10.14882/jrds.100.0.12047.0
  • Rhophilin-2遺伝子はマウス受精卵における第1分裂の細胞質分裂に関与している, 松岡 俊樹, 野老 美紀子, 申 承旭, 天野 朋子, 三谷 匡, 加藤 博己, 安斎 政幸, 岸上 哲士, 細井 美彦, 佐伯 和弘, 入谷 明, 松本 和也, The Journal of Reproduction and Development, 53, Suppl., j138, j138,   2007 09 , 10.14882/jrds.100.0.12048.0
  • マウス体細胞核移植胚における転写因子Cdx2とOct3/4の発現(Expression of transcription factor Cdx2 and Oct3/4 in mouse somatic cell nuclear transfer embryos), 西脇 恵, 三谷 匡, 安齋 政幸, 加藤 博己, 松本 和也, 佐伯 和弘, 細井 美彦, 入谷 明, 日本発生生物学会・日本細胞生物学会合同大会要旨集, 40回・59回, 81, 81,   2007 05
  • トランスジェニックマウス(Tgマウス)由来卵子及び着床前胚における母性遺伝子のプロモーター活性, ENDO NORIMASA, TSUNEMOTO KAZUNOBU, ANZAI MASAYUKI, MATSUOKA TOSHIKI, TOKORO MIKIKO, SHIN SEUNGWOOK, AMANO TOMOKO, MITANI TASUKU, KATO HIROMI, SAEKI KAZUHIRO, HOSOI YOSHIHIKO, IRITANI AKIRA, MATSUMOTO KAZUYA, 生化学, 2P-1309,   2007 , http://jglobal.jst.go.jp/detail.php?from=API&JGLOBAL_ID=200902252743397607
  • マウスMII期卵母細胞における網羅的タンパク質発現(プロテオーム)解析, TOKORO MIKIKO, KAWASUMI MIYURI, NAGAI KOHEI, IKEGAMI HARUKA, SHIN SEUNG-WOOK, MATSUOKA TOSHIKI, SATO MANABU, AMANO TOMOKO, MITANI TASUKU, KATO HIROMI, ANZAI MASAYUKI, KISHIGAMI SATOSHI, SAEKI KAZUHIRO, HOSOI YOSHIHIKO, IRITANI AKIRA, MATSUMOTO KAZUYA, 生化学, 2P-1307,   2007 , http://jglobal.jst.go.jp/detail.php?from=API&JGLOBAL_ID=200902268338533254
  • マウス体細胞核移植胚における栄養外胚葉分化に関する転写因子の発現, NISHIWAKI MEGUMI, ANZAI MASAYUKI, KATO HIROMI, HOSOI YOSHIHIKO, IRITANI AKIRA, MITANI TASUKU, 生化学, 1P-0991,   2007 , http://jglobal.jst.go.jp/detail.php?from=API&JGLOBAL_ID=200902297357245626
  • マウス体細胞核移植胚および体外受精胚におけるOct-3/4遺伝子転写調節領域のメチル化, 川澄 みゆり, 海野 祐一, 西脇 恵, 松本 和也, 安齋 政幸, 三谷 匡, 加藤 博己, 天野 朋子, 佐伯 和弘, 細井 美彦, 入谷 明, The Journal of Reproduction and Development, 52, Suppl., j75, j75,   2006 08 , 10.14882/jrds.99.0.62.0
  • 母性発現遺伝子(Histone H1oo(H1oo),Nucleoplasmin2(Npm2),Zygote arrest1(Zar1))のプロモーター領域の解析, 常本 和伸, 松本 和也, 安斎 政幸, 天野 朋子, 三谷 匡, 加藤 博己, 細井 美彦, 佐伯 和弘, 入谷 明, The Journal of Reproduction and Development, 52, Suppl., j88, j88,   2006 08 , 10.14882/jrds.99.0.87.0
  • Tetraploid embryo complementation法による遺伝子改変マウスの作出法の確立, 小澤 まどか, 寺村 岳士, 掛川 亮, 武内 大輝, 三原 敏敬, 安斎 政幸, 松本 和也, 佐伯 和弘, 佐川 典正, 細井 美彦, The Journal of Reproduction and Development, 52, Suppl., j120, j120,   2006 08 , 10.14882/jrds.99.0.152.0
  • ウサギEG細胞の樹立, 掛川 亮, 竹原 俊幸, 寺村 岳士, 小澤 まどか, 安斎 政幸, 松本 和也, 佐伯 和弘, 佐川 典正, 細井 美彦, The Journal of Reproduction and Development, 52, Suppl., j123, j123,   2006 08 , 10.14882/jrds.99.0.158.0
  • ホウレンソウ由来Δ12脂肪酸不飽和化酵素遺伝子導入マウスにおけるプロテオーム解析, SHINKAI YUSUKE, MATSUMOTO KAZUYA, AMANO TOMOKO, TAGUCHI YOSHITOMO, ANZAI MASAYUKI, MITANI TASUKU, KATO HIROMI, HOSOI YOSHIHIKO, SAEKI KAZUHIRO, SUZUKI IWANE, MURATA NORIO, IRITANI AKIRA, 日本分子生物学会年会講演要旨集, 28th, 540,   2005 11 25 , http://jglobal.jst.go.jp/detail.php?from=API&JGLOBAL_ID=200902213559909810
  • マウス初期はい特異的発現ベクターの開発, TSUNEMOTO KAZUNOBU, MATSUMOTO KAZUYA, ANZAI MASAYUKI, AMANO TOMOKO, MITANI TASUKU, KATO HIROMI, HOSOI YOSHIHIKO, SAEKI KAZUHIRO, IRITANI AKIRA, 日本分子生物学会年会講演要旨集, 28th, 703,   2005 11 25 , http://jglobal.jst.go.jp/detail.php?from=API&JGLOBAL_ID=200902216149803763
  • マウス初期はいにおけるrhophilin‐2遺伝子の機能解析, MATSUOKA TOSHIKI, HAYAKUMO MASANORI, SONO YOHEI, MATSUMOTO KAZUYA, AMANO TOMOKO, ANZAI MASAYUKI, MITANI TASUKU, KATO HIROMI, HOSOI YOSHIHIKO, SAEKI KAZUHIRO, IRITANI AKIRA, 日本分子生物学会年会講演要旨集, 28th, 703,   2005 11 25 , http://jglobal.jst.go.jp/detail.php?from=API&JGLOBAL_ID=200902246956838577
  • マウス尾部細胞,ES細胞,IVFはい及びNTはいにおけるOct‐3/4遺伝子転写調節領域のCpGメチル化, UMINO YUICHI, KAWASUMI MIYURI, MATSUMOTO KAZUYA, ANZAI MASAYUKI, AMANO TOMOKO, MITANI MASASHI, KATO HIROKI, SAEKI KAZUHIRO, HOSOI YOSHIHIKO, IRIYA AKIRA, J Reprod Dev, 51, Supplement, J65,   2005 08 25 , http://jglobal.jst.go.jp/detail.php?from=API&JGLOBAL_ID=200902259427986069
  • マウス初期胚で発現するrhophilin-2遺伝子の機能解析に関する研究, 松岡 俊樹, 園 洋平, 松本 和也, 天野 朋子, 安斎 政幸, 三谷 匡, 加藤 博己, 細井 美彦, 佐伯 和弘, 入谷 明, The Journal of Reproduction and Development, 51, Suppl., j57, j57,   2005 08
  • マウス尾部細胞,ES細胞,IVF胚及びNT胚におけるOct-3/4遺伝子転写調節領域のCpGメチル化, 海野 佑一, 川澄 みゆり, 松本 和也, 安斎 政幸, 天野 朋子, 三谷 匡, 加藤 博己, 佐伯 和弘, 細井 美彦, 入谷 明, The Journal of Reproduction and Development, 51, Suppl., j65, j65,   2005 08
  • 植物由来Δ12脂肪酸不飽和化酵素遺伝子導入マウスにおける網羅的遺伝子発現解析, SHINKAI YUSUKE, MATSUMOTO KAZUYA, AMANO TOMOKO, TAGUCHI YOSHITOMO, ANZAI MASAYUKI, MITANI TASUKU, KATO HIROMI, SUZUKI IWANE, MURATA NORIO, 日本分子生物学会年会プログラム・講演要旨集, 27th, 1009,   2004 11 25 , http://jglobal.jst.go.jp/detail.php?from=API&JGLOBAL_ID=200902205325166641
  • マウス初期はいにおけるはい性遺伝子発現機構へのDNAメチル化及びヒストンアセチル化の関与, YAMAMOTO YUMI, MATSUMOTO KAZUYA, AMANO TOMOKO, KURIHARA TAKASHI, ANZAI MASAYUKI, MITANI TASUKU, KATO HIROMI, HOSOI YOSHIHIKO, SAEKI KAZUHIRO, 日本分子生物学会年会プログラム・講演要旨集, 27th, 714,   2004 11 25 , http://jglobal.jst.go.jp/detail.php?from=API&JGLOBAL_ID=200902279039552902
  • マウス体細胞核移植はいにおける微小管の変化, KAWASUMI MIYURI, ANZAI MASAYUKI, KUNIEDA TAKANORI, KATO HIROMI, MITANI TASUKU, MATSUMOTO KAZUYA, SAEKI KAZUHIRO, HOSOI YOSHIHIKO, IRITANI AKIRA, 日本分子生物学会年会プログラム・講演要旨集, 27th, 537,   2004 11 25 , http://jglobal.jst.go.jp/detail.php?from=API&JGLOBAL_ID=200902292351200998
  • マウス初期はいにおける時計遺伝子群の発現解析, MATSUSHITA SATONORI, AMANO TOMOKO, MATSUMOTO KAZUYA, ANZAI MASAYUKI, MITANI MASASHI, KATO HIROKI, HOSOI YOSHIHIKO, SAEKI KAZUHIRO, IRIYA AKIRA, J Reprod Dev, 50, Supplement, J57,   2004 08 25 , http://jglobal.jst.go.jp/detail.php?from=API&JGLOBAL_ID=200902214844920620
  • マウス初期はいのはい性遺伝子発現機構におけるDNAメチル化及びヒストンアセチル化の関与, YAMAMOTO YUMI, MATSUMOTO KAZUYA, AMANO TOMOKO, KURIHARA TAKASHI, ANZAI MASAYUKI, MITANI MASASHI, KATO HIROKI, HOSOI YOSHIHIKO, SAEKI KAZUHIRO, J Reprod Dev, 50, Supplement, J59,   2004 08 25 , http://jglobal.jst.go.jp/detail.php?from=API&JGLOBAL_ID=200902283953789853
  • マウス初期胚における時計遺伝子群の発現解析, 松下 聡紀, 天野 朋子, 松本 和也, 安齋 政幸, 三谷 匡, 加藤 博己, 細井 美彦, 佐伯 和弘, 入谷 明, The Journal of Reproduction and Development, 50, Suppl., j57, j57,   2004 08
  • マウス初期胚で発現するrhophilin-2遺伝子と相互作用する因子の探索, 松岡 俊樹, 松本 和也, 天野 朋子, 安齋 政幸, 三谷 匡, 加藤 博己, 細井 美彦, 佐伯 和弘, 入谷 明, The Journal of Reproduction and Development, 50, Suppl., j58, j58,   2004 08
  • マウス初期胚の胚性遺伝子発現機構におけるDNAメチル化及びヒストンアセチル化の関与, 山本 由美, 松本 和也, 天野 朋子, 栗原 隆, 安齋 政幸, 三谷 匡, 加藤 博己, 細井 美彦, 佐伯 和弘, 入谷 明, The Journal of Reproduction and Development, 50, Suppl., j59, j59,   2004 08
  • 植物由来脂肪酸不飽和化酵素を発現させたトランスジェニックマウスにおける遺伝子挿入部位の解析, 新海 雄介, 湯浅 一之, 桐本 真治, 松本 和也, 天野 朋子, 安斎 政幸, 三谷 匡, 加藤 博己, 田口 善智, 細井 美彦, 佐伯 和弘, 入谷 明, The Journal of Reproduction and Development, 50, Suppl., j94, j94,   2004 08
  • ウシ体細胞クローン胚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, 安齋 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集, 103回, 112, 112,   2004 03
  • Correlation between follicle size and quality of oocytes from the superovulated cynomolgus monkey, Reproduction, Fertility and Development, 16, 1, 289,   2004 01
  • Short-Term Storage and Transport at Cold Temperatures of 2-Cell Mouse Embryos Produced by Cryopreserved Sperm, Toru Takeo, Tomoko Kondo, Yukie Haruguchi, Kiyoko Fukumoto, Yoshiko Nakagawa, Yumi Takeshita, Yuko Nakamuta, Shuuji Tsuchiyama, Norihiko Shimizu, Takanori Hasegawa, Motohito Goto, Hitoshi Miyachi, Masayuki Anzai, Rie Fujikawa, Koji Nomaru, Takehito Kaneko, Yoshiaki Itagaki, Naomi Nakagata, JOURNAL OF THE AMERICAN ASSOCIATION FOR LABORATORY ANIMAL SCIENCE, 49, 4, 415, 419,   2010 07
    Summary:At refrigerated temperatures, mouse embryos can maintain developmental ability for short periods. Previously, we succeeded in transporting vitrified and warmed 2-cell mouse embryos while maintaining developmental ability at refrigerated temperatures for 50 h. Transport of nonfrozen embryos is an easier and more useful means of exchanging genetically engineered mice between laboratories than is transport of cryopreserved embryos. Here we examined the developmental ability of transported 2-cell embryos that were produced through in vitro fertilization using cryopreserved sperm. Results show that 2-cell embryos produced by cryopreserved sperm can develop into blastocysts after cold storage for 24, 48, and 72 h. Transported 2-cell embryos produced by cryopreserved sperm yielded a favorable number of pups in all of the receiving laboratories after transport lasting 48 to 52 h. In summary, cold storage and transport of 2-cell embryos derived from cryopreserved sperm at refrigerated temperatures provides a novel means of transporting genetically engineered mice as an alternative to the transport of cryopreserved embryos and sperm.
  • Oocyte Activation in Mice Using Strontium with Calcium-Selective Chelators, Yoshiko Tsujimoto, Satoshi Kishigami, Toshiyuki Takehara, Masayuki Anzai, Tasuku Mitani, Hiromi Kato, Tomoko Amano, Kazuya Matsumoto, Kazuhiro Saeki, Akira Iritani, Yoshihiko Hosoi, BIOLOGY OF REPRODUCTION, 191, 191,   2009
  • Viability and DNA fragmentation of mouse frozen bone marrow cells without cryoprotectant and its availability for nuclear donor in somatic cell nuclear transfer, H. Kato, A. Nakao, M. Nishiwaki, M. Anzai, T. Mitani, A. Iritani, REPRODUCTION IN DOMESTIC ANIMALS, 43, 191, 192,   2008 07
  • Expression of the genes implicated in the development of the trophoblast lineage in mouse somatic cell nuclear transfer embryos, T. Mitani, M. Nishiwaki, M. Morita, K. Moriki, Y. Nishiyama, H. Kato, M. Anzai, A. Iritani, REPRODUCTION IN DOMESTIC ANIMALS, 43, 194, 194,   2008 07
  • FERTILITY IN HOMOZYGOUS TRANSGENIC RATS BEARING ANTISENSE RAT GROWTH-HORMONE GENE, K MATSUMOTO, M ANZAI, N NAKAGATA, K MIYATA, E SATO, Y TOYODA, BIOLOGY OF REPRODUCTION, 52, 142, 142,   1995
  • PRODUCTION OF TRANSGENIC MICE FROM IN-VITRO FERTILIZED-EGGS CRYOPRESERVED BY ULTRARAPID FREEZING, M ANZAI, N NAKAGATA, K MATSUMOTO, T ISHIKAWA, Y TAKAHASHI, K MIYATA, EXPERIMENTAL ANIMALS, 43, 3, 445, 448,   1994 07
    Summary:In vitro fertilized mouse eggs (C57BL/6N), followed by ultrarapid freezing were used for production of transgenic mice by microinjection of the chicken beta-actin promoter-driven the firefly luciferase cDNA (beta act-Luc). Following micromanipulation, the survival rates of the cryopreserved eggs and of the fresh in vitro fertilized eggs (control) were 70.8% (131/185) and 71.9% (159/221), respectively. After transferring them into oviducts of psudopregnant recipients on Day 1, 13.6% (17/125) of the cryopreserved eggs developed to live offspring and 14.1% (21/149) of fresh eggs did so. It was confirmed by Southern blotting analysis that each two transgenic mice were produced from the cryopreserved eggs (12%, 2/17) and the fresh eggs (10%, 2/21). All of transgenic mice produced from both eggs showed the expression of the luciferase gene. These results indicate that the in vitro fertilized eggs cryopreserved by ultrarapid freezing, can be, easily and conveniently, used for generation of transgenic mice.
  • CRYOPRESERVATION OF IN-VITRO FERTILIZED EMBRYOS FROM TRANSGENIC RAT BY ULTRARAPID FREEZING, M ANZAI, N NAKAGATA, K MATSUMOTO, A TAKAHASHI, Y TAKAHASH, K MIYATA, EXPERIMENTAL ANIMALS, 43, 2, 247, 250,   1994 04
    Summary:We cryopreserved pronuclear stage-embryos from the Transgenic rats produced by introducing the rat GH antisense transgene, by ultrarapid freezing. The embryos were obtained by fertilization in vitro with the sperm of homozygous males for the transgene. After thawing of a part of cryopreserved embryos (97 embryos), the 40 embryos (41%) were morphologically normal, and were transferred to oviducts of recipients. All four live young (10%) obtained (one male and three females) preserved the transgene and exhibited dwarfism. We also produced the homozygous transgenic rats after brother-sister mating of those young. These results demonstrated the possibility of the cryopreservation of embryos by ultrarapid freezing for sustaining transgenic rat line.
  • CRYOPRESERVATION OF TRANSGENIC MOUSE EMBRYOS BY ULTRARAPID FREEZING, M ANZAI, N NAKAGATA, K MATSUMOTO, A TAKAHASI, Y TAKAHASI, K MIYATA, EXPERIMENTAL ANIMALS, 42, 3, 467, 470,   1993 07
    Summary:Two transgenic mice lines were produced by introducing the rat GH antisense transgene and the chicken beta-actin promoter/firefly luciferase hybrid gene in our labolatory. We cryopreserved the transgenic embryos, obtained by fertilization in vitro with the sperm of hemizygous males for the transgene, by the ultrarapid freezing. The survival rates of cryopreserved 2-cell embryos were high (>70%) at thawing in both lines and 53% and 16% of cryopreserved 2-cell embryos, respectively, developed to live young after transferring to oviducts of recipients. Each transgene was detected at about 40% of the live young from two transgenic lines. These results indicated that the cryopreservation of embryos by ultrarapid freezing was valuable for sustaining transgenic mouse lines without genetic contaminations.
  • CRYOPRESERVATION OF SPERMATOZOA OF A TRANSGENIC MOUSE, N NAKAGATA, K MATSUMOTO, M ANZAI, A TAKAHASHI, Y TAKAHASHI, Y MATSUZAKI, K MIYATA, EXPERIMENTAL ANIMALS, 41, 4, 537, 540,   1992 10
    Summary:Spermatozoa of a homozygous tranasgenic mouse, in which the firefly luciferase gene was expressed under the control of beta-actin promoter, were frozen at -196-degrees-C. One fourth of the frozen sperm was later thawed and used for in vitro fertilization. Thirty-six of 65 oocytes (55.4%) developed to the 2-cell stage. All the 2-cell embryos were transferred to the oviducts of pseudopregnant recipients and 23 young (63.9%, 23/36) were born. All of young analyzed carried the transgene and showed the luciferase gene expression.

Research Grants & Projects

  • Ministry of Education, Culture, Sports, Science and Technology, Grants-in-Aid for Scientific Research(基盤研究(C)), Application to somatic cell nuclear transfer technology and the development of advanced reproductive technology engineering corresponding to the conservation of mouse genetic resources
  • Ministry of Education, Culture, Sports, Science and Technology, Grants-in-Aid for Scientific Research(新学術領域研究(研究課題提案型)), Development of genetically engineered animals by use of non-B DNA structure for highly and stably expression of transgene