Clonal origin of germ cell colonies after spermatogonial transplantation in mice
Clonal origin of germ cell colonies after spermatogonial transplantation in mice
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DOI：
10.1095/biolreprod.106.051193
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发表时间：
2006-07-01
影响因子：
3.6
通讯作者：
Shinohara, Takashi
中科院分区：
文献类型：
--
作者：
Kanatsu-Shinohara, Mito;Inoue, Kimiko;Shinohara, Takashi
精子发生起源于少量精原干细胞，这些干细胞可以在移植到不育受体睾丸后重新启动精子发生并产生生殖细胞集落。尽管之前的几项研究表明生殖细胞集落的单细胞起源，但仅提供了间接证据。在这项研究中，我们使用逆转录病毒测试了克隆起源假说，逆转录病毒可以特异性标记单个精原干细胞。精原干细胞在体外用表达增强型绿色荧光蛋白的逆转录病毒感染，然后移植到不育的受体小鼠中。从单个菌落中回收活的单倍体生殖细胞，并将其显微注射到卵中以产生后代。五个群体总共产生了 45 个后代，其中 23 个（51%）后代是转基因的。 Southern印迹分析表明，单菌落的转基因后代携带共同的整合位点，不同菌落的转基因后代的整合位点有所不同。这些结果提供了生殖细胞集落由单个精原干细胞发育而来的证据。
Spermatogenesis originates from a small number of spermatogonial stem cells that can reinitiate spermatogenesis and produce germ cell colonies following transplantation into infertile recipient testes. Although several previous studies have suggested a single-cell origin of germ cell colonies, only indirect evidence has been presented. in this investigation, we tested the clonal origin hypothesis using a retrovirus, which could specifically mark an individual spermatogonial stem cell. Spermatogonial stem cells were infected in vitro with an enhanced green fluorescence protein-expressing retrovirus and subsequently transplanted into infertile recipient mice. Live haploid germ cells were recovered from individual colonies and were microinjected into eggs to create offspring. In total, 45 offspring were produced from five colonies, and 23 (51%) of the offspring were transgenic. Southern blot analysis indicated that the transgenic offspring from the single colony carried a common integration site, and the integration site was different among the transgenic offspring from different colonies. These results provide evidence that germ cell colonies develop from single spermatogonial stem cells.