WCU Biomodulation Major, Department of Agricultural Biotechnology and Research Institute for Agriculture and Life Sciences, Seoul National University, Seoul, Korea.
In Vitro Cell Dev Biol Anim. 2010 Dec;46(10):828-33. doi: 10.1007/s11626-010-9361-8. Epub 2010 Nov 6.
The production of chimeric birds is an important tool for the investigation of vertebrate development, the conservation of endangered birds, and the development of various biotechnological applications. This study examined whether gamma (γ)-irradiation depletes endogenous primordial germ cells and enhances the efficiency of somatic chimerism in chickens. An optimal irradiation protocol for stage X embryos was determined after irradiation at various doses (0, 100, 300, 500, 600, 700, and 2,000 rad). Exposure to 500 rad of γ-irradiation for 73 s significantly decreased the number of primordial germ cells (P < 0.0001). Somatic chimera hatchlings were then produced by transferring blastodermal cells from a Korean Oge into either an irradiated (at 500 rad) or intact stage X White Leghorn embryo. An analysis of feather color pattern and polymerase chain reaction-based species-specific amplification of various tissues of the hatchlings confirmed chimerism in most organs of the chick produced from the irradiated recipient; a lesser degree of chimerism was observed in the non-irradiated control recipient. In conclusion, the exposure of chick embryos to an optimized dose of γ-irradiation effectively depleted germ cells and yielded greater somatic chimerism than non-irradiated control embryos. This technique can be applied to interspecies reproduction or the production of transgenic birds.
嵌合鸟的产生是研究脊椎动物发育、保护濒危鸟类和开发各种生物技术应用的重要工具。本研究探讨了 γ-射线辐射是否会耗尽内源性原始生殖细胞并提高鸡的体细胞核移植嵌合效率。在不同剂量(0、100、300、500、600、700 和 2000 拉德)下对 X 期胚胎进行辐射后,确定了 X 期胚胎的最佳辐射方案。暴露于 500 拉德的 γ-射线辐射 73 秒可显著减少原始生殖细胞的数量(P < 0.0001)。然后,将来自韩国 Oge 的胚泡细胞转移到经 500 拉德辐射或完整 X 期白来亨鸡胚胎中,产生体细胞核移植嵌合雏鸡。对雏鸡的羽毛颜色模式和聚合酶链反应(PCR)的各种组织的种特异性扩增分析证实,大多数来自辐射受体的雏鸡的器官中存在嵌合现象;在未辐射的对照受体中观察到较低程度的嵌合现象。总之,将鸡胚胎暴露于优化剂量的 γ-射线辐射可有效耗尽生殖细胞,并产生比未辐射对照胚胎更高的体细胞核移植嵌合率。这项技术可应用于种间繁殖或转基因鸟的生产。