Introduction of foreign primordial germ cells into quail and its expression in the next generation.

外来原始生殖细胞引入鹌鹑及其在下一代中的表达。

• 批准号: 06660351
• 负责人: ONO Tamao
• 金额: $ 1.28万
• 依托单位: Shinshu University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (C)
• 财政年份: 1994
• 资助国家: 日本
• 起止时间: 1994 至 1995
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-06660351/
• 关键词: Quail; Chicken; Embryonic development; Transgenesis; Primordial germ cells; Blastoderm; Embryo culture; gonad

Production of offspring derived from transferred foreign primordial germ cells (PGCs) will be highly attractive strategy for genetic improvement of domestic animals including poultry. In this study we transfused circulating PGCs of quail embryos to chick embryos and vice versa. Treated embryos were cultured in vitro in surrogate chicken egg-shells until reached to stage 29 of H&H and then observed the population of foreign PGCs settled in the embryonic gonads of host animals. The endogenous and exogenous PGCs were immunohistochemically stained with chick PGCs-positive 2C9 and quail PGCs-positive QCR1 monoclonal antibodies. For double immunohistochemistry paraffin sectioned embryonic gonads were treated with 2C9, visualized with DAB,removed the antibody from the antigen with 0.1M glycine-HC1 (pH2.2) , treated with QCR1 and visualized with 1-cloro-4-naphthol. Transferred quail PGCs were settled in the gonads of all the surviving chick embryos (5/10) and shared 3-19% of total population of PGCs in the gonad. In the reverse case, transfer of chick PGCs to quail embryos, chick PGCs were also settled in the gonads of all the surviving embryos (8/10) and shared 1-10% of of total population of PGCs.

利用外源原始生殖细胞（PGCs）生产后代将是包括家禽在内的家畜遗传改良的极具吸引力的策略。在本研究中，我们将鹌鹑胚胎的循环PGCs输注到鸡胚胎中，反之亦然。将处理后的胚胎放在代孕鸡的蛋壳中体外培养，直到第29期，然后观察外来PGCs在宿主动物的胚胎性腺中定居的情况。用鸡PGCs阳性的2C9和鹌鹑PGCs阳性的QCR1单克隆抗体对内源性和外源性PGCs进行免疫组织化学染色。双免疫组织化学切片胚胎性腺用2C9处理，DAB显像，用0.1M甘氨酸- hc1 （pH2.2）去除抗原上的抗体，用QCR1处理，1-氯-4-萘酚显像。移植的鹌鹑PGCs分布在所有存活的鸡胚的生殖腺中（5/10），占生殖腺PGCs总数的3-19%。相反，将雏鸡PGCs移植到鹌鹑胚胎中，雏鸡PGCs也定居在所有存活胚胎的性腺中（8/10），占总数的1-10%。

项目成果

Ono,T.et al.,: "Production of quail chimeras by transfer of early blastoderm cells : plumage chimeras and a germline chimera without plumage mixture." Japanese Poultry Science. 32. 252-256 (1995)

Ono,T.等人：“通过转移早期胚层细胞生产鹌鹑嵌合体：羽毛嵌合体和没有羽毛混合物的种系嵌合体。”

Ono,T.et al.,: "Culture of naked quail(conturnix coturnix japonica)ova in vitro for avian transgenesis: Culture from the single cell stage to hatching with pH adjusted chicken thick albumen." Comparative Biochemistry and Physiology. 113A(in press). (1996)

Ono,T.等人：“体外培养裸鹌鹑（conturnix coturnix japonica）进行禽类转基因：用调节 pH 值的鸡浓蛋白从单细胞阶段培养到孵化。”

Ono, T., Murakami, T., Tanabe, Y., Mizutani, M., Mochii, M.and Eguchi, G.: "Culture of naked quail (coturnix coturnix japonica) ova in vitro for avian transgenesis : Culture from the single-cell stage to hatching with pH-adjusted chicken thick albumen." C

小野，T.，村上，T.，田边，Y.，水谷，M.，Mochii，M.和江口，G.：“体外培养裸鹌鹑（coturnix coturnix japonica）卵用于禽类转基因：来自

Ono,T.et al.,: "Gene transfer into circulating primordial germ cells of quail embryos." Experimental Animals. 44. 275-278 (1995)

Ono,T.等人：“将基因转移到鹌鹑胚胎的循环原始生殖细胞中。”

Ono, T., Muto, S., Matsumoto, T., Mochii, M.and Eguchi, G.: "Gene transfer into circulating primordial germ cells of quail embryos." Experimental Animals. 44. 275-278 (1995)

Ono, T.、Muto, S.、Matsumoto, T.、Mochii, M. 和 Eguchi, G.：“基因转移到鹌鹑胚胎的循环原始生殖细胞中。”