受体胚龄对鸡原始生殖细胞移植后归巢的影响

【目的】探究受体胚龄对鸡原始生殖细胞(Primordial germ cell,PGC)移植后归巢的影响,确定最佳回注时间窗口,为提高转基因鸡的生产效率提供参考依据。【方法】从广西黄羽鸡胚分离获得PGC,体外培养并进行转基因操作后,移植回注到不同胚龄的受体鸡胚中,移植5 d后分离性腺拍照观察,对比受体鸡胚孵育至14HH~17HH时对PGC迁移归巢到性腺的影响。【结果】分别在14HH~17HH胚龄时移植回注PGC,移植5 d后的鸡胚死亡率为30.0%~60.9%,但不同移植胚龄间的差异不显著(P〉0.05)。根据鸡胚性腺中的GFP阳性细胞数可划分为6个等级(I~VI),且随着移植胚龄的增加,PG...

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Published in南方农业学报 Vol. 47; no. 6; pp. 1014 - 1018
Main Author 陈美娟 陈东阳 谢龙 陆振萍 杨蒙蒙 莫丽芬 卢克焕 陆阳清
Format Journal Article
LanguageChinese
Published 广西大学 亚热带农业生物资源保护与利用国家重点实验室/广西高校动物繁殖与动物生物技术重点实验室,南宁,530005 2016
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ISSN2095-1191
DOI10.3969/j:issn.2095-1191.2016.06.1014

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Summary:【目的】探究受体胚龄对鸡原始生殖细胞(Primordial germ cell,PGC)移植后归巢的影响,确定最佳回注时间窗口,为提高转基因鸡的生产效率提供参考依据。【方法】从广西黄羽鸡胚分离获得PGC,体外培养并进行转基因操作后,移植回注到不同胚龄的受体鸡胚中,移植5 d后分离性腺拍照观察,对比受体鸡胚孵育至14HH~17HH时对PGC迁移归巢到性腺的影响。【结果】分别在14HH~17HH胚龄时移植回注PGC,移植5 d后的鸡胚死亡率为30.0%~60.9%,但不同移植胚龄间的差异不显著(P〉0.05)。根据鸡胚性腺中的GFP阳性细胞数可划分为6个等级(I~VI),且随着移植胚龄的增加,PGC迁移效率呈下降趋势。14HH胚龄移植的鸡胚性腺GFP阳性细胞集中在V级(40.0%),15HH胚龄移植的集中在III级(42.1%),16HH和17HH胚龄移植的则集中在II级(45.4%和54.5%),GFP阳性细胞含量最高等级(VI级)仅出现在14HH胚龄移植的鸡胚性腺中。【结论】随着受体胚龄的增加,PGC迁移效率呈下降趋势,但鉴于实际操作的难易程度,确定PGC移植回注的最佳时间窗口为14HH鸡胚(孵育50 h)。
Bibliography:CHEN Mei-juan, CHEN Dong-yang, XIE Long, LU Zhen-ping, YANG Meng-meng, MO Li-fen, LU Ke-huan, LU Yang-qing(State Key Laboratory for Conservation and Utilization of Subtropical Agro-bioresources, Guangxi University/Guangxi High Education Laboratory for Animal Reproduction and Biotechnology, Nanning 530005, China)
45-1381/S
[Objective]The present experiment was conducted to investigate effect of recipient embryo age on homing of chicken primordial germ cell(PGC ) after transplantation and determine optimal reinjection time window, in order to provide basis for productivity of transgenic chicken. [ Method ] PGCs were isolated from embryos of native yellow feather chicken in Guangxi, and cultured in vitro, then genetically modified, finally transferred back into recipient embryos. On the 5th days after transplantation, the gonads of recipient embryos were isolated and imaged to compare effect of recipient embryo age (lgHH-17HH) on PGC migration to gonads. [ Result ]The mortality rate of embryo ranged from 30.0% to
ISSN:2095-1191
DOI:10.3969/j:issn.2095-1191.2016.06.1014