Depletion of Primordial Germ Cells (PGCs) by X-irradiation to Extraembryonic Region of Chicken Embryos and Expression of Xenotransplanted Quail PGCs
Depletion of Primordial Germ Cells (PGCs) by X-irradiation to Extraembryonic Region of Chicken Embryos and Expression of Xenotransplanted Quail PGCs
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DOI：
10.2141/jpsa.46.136
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发表时间：
2009-04-01
影响因子：
1.5
通讯作者：
Ono, Tamao
中科院分区：
文献类型：
--
作者：
Atsumi, Yusuke;Yazawa, Shigenobu;Ono, Tamao
通过移植原始生殖细胞（PGCs）产生生殖系嵌合体需要将供体的PGCs掺入受体胚胎的性腺组织中。我们研究了软X射线照射与铅（12-3 × 0.25毫米，类似于0.1克）屏蔽的应用程序，以适当的鸡鹌鹑生殖系嵌合体的生产胚胎的效用。在13 - 17期照射120 s（类似于7.2戈伊）期间屏蔽的鸡胚显示出35%（106/301）的孵化率。而未屏蔽胚胎的孵化率为26%（27/105）。在第13期照射的胚胎中，有或无屏蔽的性腺PGCs的相对数量分别为13%和5%。分别为未辐照对照的值。在第13或14阶段有或无屏蔽照射的鸡胚和用含PGCs的鹌鹑胚胎血输注的鸡胚，在第30阶段的左侧性腺中每个都显示出类似于130个供体PGCs相对群体。通过Southern杂交和PCR检测，异种移植的幼鱼表现出供体来源的PGCs。因此，在第13阶段将鸡胚暴露于类似于7.2戈伊的X射线中，并将铅屏蔽物应用于胚胎本身，是耗尽内源性生殖细胞和产生鸡-鹌鹑生殖系嵌合体的可行方法。
The generation of germline chimeras by the transfer of primordial germ cells (PGCs) requires incorporation of the PGCs of the donor into the gonadal tissue of the recipient embryo. We investigated the utility of soft x-irradiation with application of a lead (12-3 x 0.25 mm, similar to 0.1 g) shield to the embryo proper for the production of chicken-quail germline chimeras. Chicken embryos shielded during irradiation for 120 s (similar to 7.2 Gy) at stages 13 to 17 showed a hatchability of 35% (106/301). whereas the hatchability of unshielded embryos was 26% (27/105). The relative Population of gonadal PGCs Lit stage 30 for embryos irradiated at stage 13 with or Without shielding was 13 and 5%. respectively, of the value for nonirradiated controls. Chicken embryos irradiated at stages 13 or 14 with or Without shielding and transfused with quail embryonic blood containing PGCs each exhibited similar to 130 relative population of donor PGCs in the left gonad at stage 30. Xenotransplanted hatchlings exhibited donor-derived PGCs as detected by Southern hybridization and PCR. Exposure of chicken embryos to similar to 7.2 Gy of x-radiation at stage 13 with the application of a lead shield to the embryo proper is thus a feasible approach to depletion of endogenous germ cells and the production of chicken-quail germline chimeras.