Restoration of various species of Galliformes using chickens

使用鸡恢复各种鸡形目动物

• 批准号: 13460132
• 负责人: ONO Tamao
• 金额: $ 10.82万
• 依托单位: Shinshu University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 2001
• 资助国家: 日本
• 起止时间: 2001 至 2003
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-13460132/
• 关键词: chicken; quail; Blue-breasted quail; embryogenesis; primordial germ cells; chimera; PCR; in situ hybridization

In order to rescue the endangered species, and production and preservation of variable breeds in bird, interspecific transfers of primoridal germ cells (PGCs) have been performed. We developed the protocol to obtain a large amount of PGCs from gonads of chicken and quail embryos using PGC specific monoclonal antibodies (anti-SSEA1 and QCR1) and magnetism activated cell sorting (MACS). MACS treatment greatly increased the population ratio of avian PGCs in gonadal cells, resulting improved gonadal migration in recipient embryos. We have worked toward practical use of PCR and in situ hybridization to identify the species specific cell (genome) type among chickens, quail and blue-breasted quail. PGCs of chick embryos were transferred into quail embryos, and the chicken-genome-specific PCR product was observed in the semen of chick-PGC-transferred chimeric quail. We made a progeny test. Females of the putative chimeras were artificially inseminated with chicken semen. We obtained hatchlings but PCR analysis showed that they were hybrid between chicken sperm and donor derived quail ovum. Silky PGCs were introduced to White Leghorn chickens and the prpgeny test is underway.

为了拯救濒危物种，培育和保存鸟类的变异品种，进行了原生生殖细胞（PGCs）的种间转移。我们开发了利用PGC特异性单克隆抗体（抗ssea1和QCR1）和磁激活细胞分选（MACS）从鸡和鹌鹑胚胎性腺中获得大量PGCs的方案。MACS处理大大增加了禽类生殖腺细胞中PGCs的种群比例，从而改善了受体胚胎的性腺迁移。我们已经在鸡、鹌鹑和蓝胸鹌鹑的物种特异性细胞（基因组）类型的鉴定中开展了实际应用PCR和原位杂交的工作。将鸡胚的PGCs转移到鹌鹑胚中，在鸡胚- PGCs转移的嵌合鹌鹑精液中观察到鸡基因组特异性PCR产物。我们做了一个子代测试。假定的嵌合体的雌性用鸡精液人工授精。我们得到了雏鸟，但PCR分析表明它们是鸡精子和供体鹌鹑卵子的杂交产物。丝质PGCs已被引入白来港鸡，目前正在进行子代试验。

项目成果

Obata, K, Koide, M., Nagata, K., Iio, A., Yazawa, S., Ono, T., Yamada, Y., Tuan, R.S., Yokota, M.: "Role of FK506-binding protein 12 in development of the chick embryonic heart."Biochemical and Biophysical Research Communications. 283(3). 621-626 (2001)

Obata，K，Koide，M.，Nagata，K.，Iio，A.，Yazawa，S.，Ono，T.，Yamada，Y.，Tuan，R.S.，Yokota，M.：“FK506 结合蛋白的作用

Li, H.C., Kagami, H., Matsui, K., Ono, T.: "Restriction of proliferation of primordial germ cells by the irradiation of Japanese quail embryos with soft X-rays."Comparative Biochemistry and Physiology Part A. 130(1). 133-140 (2001)

Li，H.C.，Kagami，H.，Matsui，K.，Ono，T.：“用软X射线照射日本鹌鹑胚胎来限制原始生殖细胞的增殖。”比较生物化学和生理学第A.130部分（

Obata, K., Koide, M., Nagata, K., Iio, A., Yazawa, S., Ono, T., Sasaki, S., Yamada, Y., Tuan, R.S., Yokota, M.: "Effects of FK506 and rapamycin on formation of the neural tube in chick embryos."Animal Science Journal. 73(3). 229-234 (2002)

Obata, K.、Koide, M.、Nagata, K.、Iio, A.、Yazawa, S.、Ono, T.、Sasaki, S.、Yamada, Y.、Tuan, R.S.、Yokota, M.：“

Hrabia, A.(筆頭著者): "Fertilization and development of quail oocytes after intracytoplasmic sperm injection"Biology of Reproduction. 69(5). 1651-1657 (2003)

Hrabia, A.（第一作者）：“胞浆内精子注射后鹌鹑卵母细胞的受精和发育”《生殖生物学》69(5) 1651-1657 (2003)。

Ono, T.: "Ex ovo culture of quail embryos and its application for embryo manipulation."Animal Science Journal. 72(5). 361-371 (2001)

Ono, T.：“鹌鹑胚胎的卵外培养及其在胚胎操作中的应用。”动物科学杂志。