Germline Replacement by Transfer of Primordial Germ Cells into Partially Sterilized Embryos in the Chicken
Germline Replacement by Transfer of Primordial Germ Cells into Partially Sterilized Embryos in the Chicken
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DOI：
10.1095/biolreprod.110.083923
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发表时间：
2010-07-01
影响因子：
3.6
通讯作者：
Tagami, Takahiro
中科院分区：
文献类型：
--
作者：
Nakamura, Yoshiaki;Usui, Fumitake;Tagami, Takahiro
我们报告了一种新技术，可以用鸡体内的供体生殖细胞几乎完全替代受体种系。在孵化前将溶解在缓释乳液中的白消安注射到受精蛋的蛋黄中。研究发现，100 μg 的剂量在减少胚胎性腺中内源性原始生殖细胞 (PGC) 数量（对照数量的 0.6%）和孵化率（36.4%）方面提供了最佳结果。这用于制备部分绝育的胚胎，作为外源 PGC 移植的受体。免疫组织化学分析显示，白消安处理胚胎中供体 PGC 的比例显着高于对照组（98.6% vs. 6.4%）。遗传交叉测试分析显示，白消安处理鸡的种系传播率显着高于对照组（99.5% vs. 6.0%）。在 11 个嵌合体中，有 7 个仅产生来自供体的后代，这表明这些嵌合体在受体的性腺中仅产生来自供体的配子。这种新颖的种系替换技术为研究种系分化、产生转基因个体和保护鸟类遗传资源提供了强大的工具。
We report a novel technique for almost complete replacement of the recipient germline with donor germ cells in the chicken. Busulfan solubilized in a sustained-release emulsion was injected into the yolk of fertile eggs before incubation. A dose of 100 mu g was found to provide the best outcome in terms of reducing the number of endogenous primordial germ cells (PGCs) in embryonic gonads (0.6% of control numbers) and hatchability (36.4%). This was applied for preparing partially sterilized embryos to serve as recipients for the transfer of exogenous PGCs. Immunohistochemical analysis showed that the proportion of donor PGCs in busulfan-treated embryos was significantly higher than in controls (98.6% vs. 6.4%). Genetic cross-test analysis revealed that the germline transmission rate in busulfan-treated chickens was significantly higher than in controls (99.5% vs. 6.0%). Of 11 chimeras, 7 produced only donor-derived progenies, suggesting that these produced only donor-derived gametes in the recipient's gonads. This novel germline replacement technique provides a powerful tool for studying germline differentiation, for generating transgenic individuals, and for conserving genetic resources in birds.