Improvement of microinsemination techniques with primary spermatocytes of mice based on live-cell imaging analysis

• Project/Area Number: 23500507
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: Laboratory animal science
• Research Institution: The Institute of Physical and Chemical Research
• Principal Investigator: OGONUKI Narumi 独立行政法人理化学研究所, バイオリソースセンター, 専任技師 (40373287)
• Project Period (FY): 2011 – 2013
• Project Status: Completed (Fiscal Year 2013)
• Budget Amount: ¥4,940,000 (Direct Cost: ¥3,800,000、Indirect Cost: ¥1,140,000); Fiscal Year 2013: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000); Fiscal Year 2012: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000); Fiscal Year 2011: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000)
• Keywords: マウス / 一次精母細胞 / 顕微授精 / ライブセルイメージング / 発生工学

Research Abstract

Microinsemination is a technique to deliver male germ cells directly into oocytes, and is a very effective technique to fertilize oocytes consistently, even male germ cells that are unable to fertilize oocytes under conventional in vovo and in vitro conditions. Although offspring can be obtained by primary spermatocyte injection in mice, its efficiency is still low (2-4%) compared with that by spermatid (30%) or by spermatozoa (40-50%). To know whether the poor full-term development, primary spermatocyte injected oocytes were monitored by live-cell imaging techniques. The gap of the chromosome division timing was observed between male and female chromosomes. And a histone, which are globally deacetylated in mammalian oocytes during meiosis, was remained acetylated in primary spermatocyte injected oocytes. Thus, inadequate histone deacetylation may be caused chromosome abnormalities and low oocyte developments.

Research Products

• [Journal Article] Oligo-astheno-teratozoospermia in mice lacking ORP4, a sterol-binding protein in the OSBP-related protein family (2014)
  • Author(s): Udagawa O, Ito C, Ogonuki N, Sato H, Lee S, Tripuvanuntakul P, Ichi I, Uchida Y, Nishimura T, Murakami M, Ogura A, Inoue T, Toshimori K, Arai H
  • Journal Title: Genes Cells Volume: 19 Pages: 13-27
  • Peer Reviewed
• [Journal Article] RNA sequencing-based identification of aberrant imprinting in cloned mice (2014)
  • Author(s): Okae H, Matoba S, Nagashima T, Mizutani E, Inoue K, Ogonuki N, Chiba H, Funayama R, Tanaka S, Yaegashi N, Nakayama K, Sasaki H, Ogura A, Arima T
  • Journal Title: Hum Mol Genet Volume: 23 Pages: 992-1001
  • Peer Reviewed
• [Journal Article] A heterozygous mutation of GALNTL5 affects male infertility with impairment of sperm motility (2014)
  • Author(s): Takasaki N, Tachibana K, Ogasawara S, Matsuzaki H, Hagiuda J, Ishikawa H, Mochida K, Inoue K, Ogonuki N, Ogura A, Noce T, Ito C, Toshimori K, Narimatsu H.
  • Journal Title: Proc Natl Acad Sci USA Volume: 111 Issue: 3 Pages: 1120-1125
  • DOI: 10.1073/pnas.1310777111
  • Peer Reviewed
• [Journal Article] RNA sequencing-based identification of aberrant imprinting in cloned mice. (2014)
  • Author(s): Okae H, Matoba S, Nagashima T, Mizutani E, Inoue K, Ogonuki N, Chiba H, Funayama R, Tanaka S, Yaegashi N, Nakayama K, Sasaki H, Ogura A, Arima T.
  • Journal Title: Hum Mol Genet. Volume: 23 Issue: 4 Pages: 992-1001
  • DOI: 10.1093/hmg/ddt495
  • Peer Reviewed
• [Journal Article] Oligo-astheno-teratozoospermia in mice lacking ORP4, a sterol-binding protein in the OSBP-related protein family. (2014)
  • Author(s): Udagawa O, Ito C, Ogonuki N, Sato H, Lee S, Tripvanuntakul P, Ichi I, Uchida Y, Nishimura T, Murakami M, Ogura A, Inoue T, Toshimori K, Arai H.
  • Journal Title: Genes Cells. Volume: 19 Issue: 1 Pages: 13-27
  • DOI: 10.1111/gtc.12105
  • Peer Reviewed
• [Journal Article] Regulation of pluripotency in male germline stem cells by Dmrt1 (2013)
  • Author(s): Takashima S, Hirose M, Ogonuki N, Ebisuya M, Inoue K, Kanatsu-Shinohara M, Tanaka T, Nishida E, Ogura A, Shinohara T
  • Journal Title: Genes Dev Volume: 27 Pages: 1949-1958
  • Peer Reviewed
• [Journal Article] ROS are required for mouse spermatogonial stem cell self-renewal (2013)
  • Author(s): Morimoto H, Iwata K, Ogonuki N, Inoue K, Ogura A, Kanatsu-Shinohara M, Morimoto T, Yabe-Nishimura C, Shinohara T
  • Journal Title: Cell Stem Cell Volume: 12 Pages: 774-786
  • NAID: 120005285490
  • Peer Reviewed
• [Journal Article] t-SNARE Syntaxin2 (STX2) is implicated in intracellular transport of sulfoglycolipids during meiotic prophase in spermatogenesis (2013)
  • Author(s): Fujiwara Y, Ogonuki N, Inoue K, Ogura A, Handel MA, Noguchi J, Kunieda T
  • Journal Title: Biol Reprod88 Volume: 141 Pages: 1-9
  • Peer Reviewed
• [Journal Article] Mouse cloning using a drop of peripheral blood (2013)
  • Author(s): Kamimura S, Inoue K, Ogonuki N, Hirose M, Oikawa M, Yo M, Ohara O, Miyoshi H, Ogura A
  • Journal Title: Biol Reprod 89 Volume: 24 Pages: 1-6
  • Peer Reviewed
• [Journal Article] RNAi-mediated knockdown of Xist does not rescue the impaired development of female cloned mouse embryos (2013)
  • Author(s): Oikawa M, Matoba S, Inoue K, Kamimura S, Hirose M, Ogonuki N, Shiura H, Sugimito M, Abe K, Ishino F, Ogura A
  • Journal Title: J Reprod Dev Volume: 59 Pages: 231-237
  • NAID: 10031184751
  • Peer Reviewed
• [Journal Article] Development of microinsemination techniques for laboratory animals and its practical applications (2013)
  • Author(s): Ogonuki, N
  • Journal Title: J. Mamm. Ova Res. Volume: 30 Pages: 159-167
  • NAID: 130004436875
  • Peer Reviewed
• [Journal Article] RNAi-mediated knockdown of Xist does not rescue the impaired development of female cloned mouse embryos. (2013)
  • Author(s): Oikawa M, Matoba S, Inoue K, Kamimura S, Hirose M, Ogonuki N, Shiura H, Sugimito M, Abe K, Ishino F, Ogura A.
  • Journal Title: J Reprod Dev Volume: vol
  • NAID: 10031184751
  • Peer Reviewed
• [Journal Article] Takashi Shinohara. Reconstitution of mouse spermatogonial stem cell niches in culture (2012)
  • Author(s): Kanatsu-Shinohara M, Inoue K, Takashima S, Takehashi M, Ogonuki N, Morimoto H, Nagasawa T, Ogura A
  • Journal Title: Cell Stem Cell Volume: 11 Pages: 567-578
  • Peer Reviewed
• [Journal Article] Testis tissue explantation cures spermatogenic failure in c-Kit ligand mutant mice (2012)
  • Author(s): Sato T, Yokonishi T, Komeya M, Katagiri K, Kubota Y, Matoba S, Ogonuki N, Ogura A, Yoshida S, Ogawa T
  • Journal Title: Proc Natl Acad Sci USA Volume: 109 Pages: 16934-16938
  • Peer Reviewed
• [Journal Article] Reconstitution of mouse spermatogonial stem cell niches in culture (2012)
  • Author(s): Kanatsu-Shinohara, M., Inoue, K., Takashima, S.,Takehashi, M.,Ogonuki, N., Morimoto, H., Nagasawa, T., Ogura, A. and Shinohara, T
  • Journal Title: Cell Stem Cell Volume: 11(4) Issue: 4 Pages: 567-578
  • DOI: 10.1016/j.stem.2012.06.011
  • Peer Reviewed
• [Journal Article] Testis tissue explantation cures spermatogenic failure in c-Kit ligand mutant mice (2012)
  • Author(s): T.Sato, T.Yokonishi, M.Komeya, K.Katagiri, Y.Kubota, S.Matoba, N.Ogonuki, A.Ogura, S.Yoshida and T.Ogawa
  • Journal Title: Proc. Natl. Acad. Sci. USA Volume: 109 Issue: 42 Pages: 16934-16938
  • DOI: 10.1073/pnas.1211845109
  • Peer Reviewed
• [Journal Article] RNAi-mediated knockdown of Xist can rescue the impaired postimplantation development of cloned mouse embryos (2011)
  • Author(s): Matoba S, Inoue K, Kohda T, Sugimioto M, Mizutani E, Ogonuki N, Nakamura T, Abe K, Nakano T, Ishino F, and Ogura A
  • Journal Title: Proc Natl Acad Sci USA Volume: 108 Pages: 20621-20626
  • Peer Reviewed
• [Journal Article] In vitro production of fertile sperm from murine spermatogonial stem cell lines (2011)
  • Author(s): T. Sato, K. Katagiri, T. Yokonishi, Y. Kubota, K. Inoue, N. Ogonuki, S. Matoba, A. Ogura, T. Ogawa
  • Journal Title: Nat Commun Volume: 2 Issue: 1 Pages: 472-472
  • DOI: 10.1038/ncomms1478
  • Peer Reviewed
• [Journal Article] Intracytoplasmic sperm injection induces transcriptome perturbation without any transgenerational effect (2011)
  • Author(s): Kohda T, Ogonuki N, Inoue K, Furuse T, Kaneda H, Suzuki T, Kaneko-Ishino T, Wakayama T, Wakana S, Ogura A, Ishino F
  • Journal Title: Biochem Biophys Res Commun Volume: 410 Pages: 282-288
  • Peer Reviewed
• [Journal Article] In vitro production of functional sperm in cultured neonatal mouse testes (2011)
  • Author(s): Sato T, Katagiri K, Gohbara A, Inoue K, Ogonuki N, Ogura A, Kubota Y, & Ogawa T
  • Journal Title: Nature Volume: 471 Pages: 504-507
  • Peer Reviewed
• [Journal Article] Birth of normal mice following round spermatid injection without artificial oocyte activation (2011)
  • Author(s): Ogonuki N, Inoue K, Ogura A
  • Journal Title: J Reprod Dev Volume: 57 Pages: 534-538
  • NAID: 10029593452
  • Peer Reviewed
• [Journal Article] Jr. Production of mouse embryonic stem cell lines from maturing oocytes by direct conversion of meiosis into mitosis (2011)
  • Author(s): Fulka H, Hirose M, Inoue K, Ogonuki N, Wakisaka N, Matoba S, Ogura A, Mosko T, Kott T Fulka, J
  • Journal Title: Stem Cells Volume: 29 Pages: 517-527
  • Peer Reviewed
• [Journal Article] Serum- and Feeder-Free Culture of Mouse Germline Stem Cells (2011)
  • Author(s): Kanatsu-Shinohara M, Inoue K, Ogonuki N, Morimoto H, Ogura A, Shinohara T
  • Journal Title: Biol. Reprod Volume: 84 Pages: 97-105
  • Peer Reviewed
• [Journal Article] RNAi-mediated knockdown of Xist can rescue the impaired postimplantation development of cloned mouse embryos. (2011)
  • Author(s): Matoba S, Inoue K, Kohda K, Sugimioto M, Mizutani E, Ogonuki N, Nakamura T, Abe K, Nakano T, Ishino F, and Ogura A.
  • Journal Title: Proc. Natl. Acad. Sci. USA Volume: 108 Pages: 20621-20626
  • Peer Reviewed
• [Journal Article] Intracytoplasmic sperm injection induces transcriptome perturbation without any transgenerational effect. (2011)
  • Author(s): Kohda T, Ogonuki N, Inoue K, Furuse T, Kaneda H, Suzuki T, Kaneko-Ishino T, Wakayama T, Wakana S, Ogura A, Ishino F.
  • Journal Title: Biochem. Biophys. Res. Commun. Volume: 410 Pages: 282-288
  • Peer Reviewed
• [Journal Article] In vitro production of functional sperm in cultured neonatal mouse testes. (2011)
  • Author(s): Sato T, Katagiri K, Gohbara A, Inoue K, Ogonuki N, Ogura A, Kubota Y, & Ogawa T.
  • Journal Title: Nature Volume: 471 Pages: 504-507
  • Peer Reviewed
• [Journal Article] Birth of normal mice following round spermatid injection without artificial oocyte activation. (2011)
  • Author(s): Ogonuki N, Inoue K, Ogura A.
  • Journal Title: J. Reprod. Dev. Volume: 57 Pages: 534-538
  • NAID: 10029593452
  • Peer Reviewed
• [Journal Article] Production of mouse embryonic stem cell lines from maturing oocytes by direct conversion of meiosis into mitosis. (2011)
  • Author(s): Fulka H, Hirose M, Inoue K, Ogonuki N, Wakisaka N, Matoba S, Ogura A, Mosko T, Kott T Fulka, J. Jr.
  • Journal Title: Stem Cells Volume: 29 Pages: 517-527
  • Peer Reviewed
• [Journal Article] Serum- and Feeder-Free Culture of Mouse Germline Stem Cells. (2011)
  • Author(s): Kanatsu-Shinohara M, Inoue K, Ogonuki N, Morimoto H, Ogura A, Shinohara T.
  • Journal Title: Biol. Reprod. Volume: 84 Pages: 97-105
  • Peer Reviewed
• [Presentation] 簡便凍結組織の精子・精細胞による受精 (2014)
  • Author(s): 越後貫成美
  • Organizer: 第32回日本受精着床学会総会・学術講演会
  • Place of Presentation: 東京、ハイアットリージェンシー東京
• [Presentation] Nuclear transfer provides a new picture of the X chromosome inactivation cycle (2013)
  • Author(s): 及川真実, ほか
  • Organizer: 26th Mouse Molecular Genetics
  • Place of Presentation: ケンブリッジ
  • Year and Date: 2013-09-20
• [Presentation] 卵子由来の Xist のインプリントは着床期マウス胚から完全に消失する (2013)
  • Author(s): 及川真実, ほか
  • Organizer: 第106回日本繁殖生物学会
  • Place of Presentation: 東京
  • Year and Date: 2013-09-12
• [Presentation] Production of cloned mice from a drop of peripheral blood (2013)
  • Author(s): 上村悟氏, ほか
  • Organizer: The 10th Annual Conference of the Asian Reproductive Biotechnology Society
  • Place of Presentation: ベトナム
  • Year and Date: 2013-08-22
• [Presentation] Xist ノックダウン処理は雌クローン胚の発生能を改善しない (2013)
  • Author(s): 及川真実, ほか
  • Organizer: 第54回日本卵子学会
  • Place of Presentation: 東京
  • Year and Date: 2013-05-26
• [Presentation] 核移植技術による Xist遺伝子のインプリント確立時期の解析 (2012)
  • Author(s): 及川真実, ほか
  • Organizer: 第35回日本分子生物学会
  • Place of Presentation: 福岡
• [Presentation] 核移植技術を利用したXist 遺伝子のインプリント確立時期の解析 (2012)
  • Author(s): 及川真実, ほか
  • Organizer: 第105回日本繁殖生物学会
  • Place of Presentation: つくば
• [Presentation] Transcriptome perturbation induced by intracytoplasmic sperm injection in the mouse (2011)
  • Author(s): 幸田尚, ほか
  • Organizer: 第34回日本分子生物学会
  • Place of Presentation: 横浜
• [Presentation] RNAi ノックダウンシステムを利用した Xist 発現抑制による体細胞クローン胚の発生能改善 (2011)
  • Author(s): 的場章悟, ほか
  • Place of Presentation: 盛岡
• [Presentation] 人為的活性化処理なしでの凍結円形精子細胞の顕微注入によるマウス産仔の作出 (2011)
  • Author(s): 越後貫成美, ほか
  • Organizer: 第58回日本実験動物学会
  • Place of Presentation: 東京
• [Presentation] 人為的卵子活性化処理なしでの凍結円形精子細胞の顕微注入によるマウス産仔の作出 (2011)
  • Author(s): 越後貫成美、井上貴美子、小倉淳郎
  • Organizer: 第58回日本実験動物学会
  • Place of Presentation: タワーホール船堀（東京）
• [Remarks]
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• [Remarks] 理化学研究所
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• [Remarks] 遺伝工学基盤後術室
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• [Remarks] 遺伝工学基盤技術室ホームページ
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