Studies on development of avian embryo after ICSI

• Project/Area Number: 19580325
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Applied animal science
• Research Institution: Nagoya University
• Principal Investigator: SHIMADA Kiyoshi Nagoya University, 生命農学研究科, 名誉教授 (40065579)
• Co-Investigator(Kenkyū-buntansha): 小野 珠乙 信州大学, 農学部, 教授 (10177264)
• Co-Investigator(Renkei-kenkyūsha): ONO Tamao 信州大学, 農学部, 教授 (10177264)
• Project Period (FY): 2007 – 2008
• Project Status: Completed (Fiscal Year 2008)
• Budget Amount: ¥3,640,000 (Direct Cost: ¥2,800,000、Indirect Cost: ¥840,000); Fiscal Year 2008: ¥1,170,000 (Direct Cost: ¥900,000、Indirect Cost: ¥270,000); Fiscal Year 2007: ¥2,470,000 (Direct Cost: ¥1,900,000、Indirect Cost: ¥570,000)
• Keywords: 細胞内精子顕微注入(ICSI) / 精子由来卵子活性化因子 / phospholipase PLCζ / cRNA / 胚盤葉 / 外来遺伝子GFP / トランスジェニックウズラ / PLCzeta / ICSI / GFP / ウズラ / 胚盤葉発生 / 顕微授精(ICSI) / phospholipase Czeta (PLCzeta) / ストロンチウム / Ca^<2+> / 発生率

Research Abstract

非ウィルス法による鳥類トランスジェニック作出は、成功率が極めて低い。よって本研究は、細胞質内精子顕微注射法(Intracytoplasmic Sperm Injection, ICSI)による遺伝子導入法を試みた。外来遺伝子としてGreen Fluorescent Protein(GFP)を単一射出精子とPLCzeta cRNAと同時に注入し、ウズラ胚にGFPを発現させるた。pCX-E GFPプラスミドとPLCzeta cRNAをウズラ未受精卵に同時に顕微注入し、24時間の体外培養(System Q1a)と48時間の追加体外培養の結果、外来遺伝子GFPを胚盤葉で発現していることが観察できた。

Research Products

• [Journal Article] Identification of differentially expressed genes involved in the regression and development of Mullerian duct of chicken (2009)
  • Author(s): Ha Y, Tsukada A, Saito N, Shimada K
  • Journal Title: International Journal of Developmental Biology 52 Pages: 1135-1141
  • Peer Reviewed
• [Journal Article] Expressionof P450arom, AMH and Erα mRNA in gonads of turkey, duck and goose within one week of age (2008)
  • Author(s): Koba N, Ohfuji T, Ha Y, Mizushima S, Tsukada A, Saito N, Shimada K.
  • Journal Title: Journal of Poultry Science 45 Pages: 215-219
  • Peer Reviewed
• [Journal Article] Developmental enhancement of intracytoplasmic sperm injection(ICSI)-generated quail embryos by phospholipase Cζ (2008)
  • Author(s): Mizushima S, Takagi S, Ono T, Atsumi Y, Tsukada A, Saito N, Shimada K.
  • Journal Title: Journal of Poultry Science 45 Pages: 152-158
  • Peer Reviewed
• [Journal Article] Profiles of mRNA expression of FOXL2, P450arom, DMRT1, AMH. P450c17, SF1, Era and AR, in relation to gonadal sex differentiation in duck embryo (2008)
  • Author(s): Koba N, Ohfuji T, Ha Y, Mizushima S, Tsukada A, Saito N, Shimada K.
  • Journal Title: Journal of Poultry Science 45 Pages: 132-138
  • Peer Reviewed
• [Journal Article] Effects of aromatase inhibitor(Fadrozole)-induced sex-reversal on gonadal differentiation and mRNA expression of P450arom, AMH and Erα in quail embryos and growth in posthaching quail (2008)
  • Author(s): Koba N, Mori M, Ha Y, Mizushima S, Tsukada A, Saito N, Shimada K
  • Journal Title: Journal of Poultry Science 45 Pages: 116-124
  • Peer Reviewed
• [Journal Article] Possible role of calcium on oocyte development after intracytoplasmic sperm injection in quail(Coturnix japonica) (2008)
  • Author(s): Mizushima S, Takagi S, Ono T, Atsumi Y, Tsukada A, Saito N, Shimada K
  • Journal Title: Journal of Experimental Zoology 307 Pages: 647-53
  • Peer Reviewed
• [Journal Article] Profiles of mRNA expression of FOXL2, P450arom, DMRT1, AMH. P450c17, SF1, ERa and AR, in relation to gonadal sex differentiation in duck embryo. (2008)
  • Author(s): Koba N, Ohfuji T, Ha Y, Mizushima S, Tsukada A, Saito N, Shimada K.
  • Journal Title: Journal of Poultry Science 45 Pages: 132-138
  • Peer Reviewed
• [Journal Article] Developmental enhancement of intracytoplasmic sperm injection (ICSI)-generated quail embryos by phospholipase Cζ (2008)
  • Author(s): Mizushima S, Takagi S, Ono T, Atsumi Y, Tsukada A, Saito N, Shimada K.
  • Journal Title: Journal of Poultry Science 45 Pages: 152-158
  • Peer Reviewed
• [Journal Article] Effects of aromatase inhibitor (Fadrozole)-induced sex-reversal on gonadai differentiation and mRNA expression of P450axm, AMH and ERa in quail embryos and growth in posthaching quail. (2008)
  • Author(s): Koba N, Mori M, Ha Y, Mizushima S, Tsukada A, Saito N, Shimada K.
  • Journal Title: Journal of Poultry Science 45 Pages: 116-124
  • Peer Reviewed
• [Journal Article] Histocompatible chicken inbred lines : Homogeneities in the major histocompatibility complex antigens of the GSP, GSN/1, PNP/DO and BM-C inbred lines assessed by hemagglutination, mixed lymphocyte reaction and skin transplantation (2007)
  • Author(s): Valdez Jr MB, Mizutani M, Fujihara A Yazawa H, Yamagata T, Shimada K, Namikawa T
  • Journal Title: Experimental Animals 56 Pages: 329-338
  • NAID: 10019776577
  • Peer Reviewed
• [Journal Article] Z-chromosome specific primers for chicken-quail hybrid blastoderm (2007)
  • Author(s): Takagi S, Ono T, Tsukada A, Atsumi Y, Mizushima S, Saito N, Shimada K.
  • Journal Title: Journal of Poultry Science 44 Pages: 209-212
  • NAID: 130004446236
  • Peer Reviewed
• [Journal Article] Fertilization and blastoderm development of quail oocytes after intracytoplasmic injection of chicken sperm bearing the W chromosome (2007)
  • Author(s): Takagi S, Ono T, Tsukada A, Atsumi Y, Mizushima S, Saito N, Shimada K
  • Journal Title: Poultry Science 86 Pages: 937-943
  • Peer Reviewed
• [Journal Article] Fertilizing ability of chicken sperm bearing W chromosome (2007)
  • Author(s): Takagi S, Tsukada A, Saito N, Shimada K.
  • Journal Title: Poultry Science 86 Pages: 730-737
  • Peer Reviewed
• [Journal Article] Possible role of calcium on oocyte development after intracytoplasmicsperm injection in quail(Conturnix japonica) (2007)
  • Author(s): Mizushima, S., et. al.
  • Journal Title: J Expt Zool 307A, Pages: 647-653
  • Peer Reviewed
• [Journal Article] Fertilizing ability of chicken sperm bearing the W chromosome (2007)
  • Author(s): Takagi, S., et. al.
  • Journal Title: Poultry Science 86 Pages: 731-738
  • Peer Reviewed
• [Journal Article] Fertilization and blastoderm development of quail oocytes after intracytoplasmic injection of chicken sperm bearing the W chromosome (2007)
  • Author(s): Takagi, S., et. al.
  • Journal Title: Poultry Science 86 Pages: 937-943
  • Peer Reviewed
• [Presentation] ウズラ卵子DNase I活性に及ぼす精子数の効果 (2009)
  • Author(s): 杉本宰甫、水島秀成、古寺敏子、島田清司、小野珠乙、木村一郎
  • Organizer: 日本家禽学会
  • Place of Presentation: 藤沢市
  • Year and Date: 2009-03-28
• [Presentation] 顕微授精ウズラ卵cRNA注入の効果 (2008)
  • Author(s): 水島秀成、島田清司
  • Organizer: 日本家禽学会
  • Place of Presentation: 水戸市
  • Year and Date: 2008-03-28
• [Presentation] Profiles of mRNA expression of sex-differentiation-related genes in relation to gonadal sexual differentiation in duck (2007)
  • Author(s): Naomi Koba, Shusei Mizushima, Akira Tsukada, Noboru Saito
  • Organizer: APPC
  • Place of Presentation: Bangkok, Thailand
  • Year and Date: 2007-02-06