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This Article Full Text Full Text (PDF) All Versions of this Article: 75/1/68    most recent biolreprod.106.051193v1 Alert me when this article is cited Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Kanatsu-Shinohara, M. Articles by Shinohara, T. Search for Related Content PubMed PubMed Citation Articles by Kanatsu-Shinohara, M. Articles by Shinohara, T. Agricola Articles by Kanatsu-Shinohara, M. Articles by Shinohara, T.

Clonal Origin of Germ Cell Colonies after Spermatogonial Transplantation in Mice¹

Horizontal Medical Research Organization,³ Department of Molecular Genetics,⁴ Center for Medical Education,⁵ Graduate School of Medicine, Kyoto University, Kyoto 606-8507, Japan RIKEN,⁶ Bioresource Center, Ibaraki 305-0074, Japan

ABSTRACT

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.

developmental biology, gametogenesis, Sertoli cells, spermatogenesis, testis

¹ Supported by the Ministry of Education, Culture, Sports, Science, and Technology (MEXT) of Japan and by grants from CREST and the Human Science Foundation (Japanese) and also supported in part by Special Coordination Funds for Promoting Science and Technology from MEXT.

² Correspondence: Mito Kanatsu-Shinohara, Horizontal Medical Research Organization, Graduate School of Medicine, Kyoto University 53 Shogoin, Kawahara-cho, Sakyo-ku, Kyoto 606-8507, Japan. FAX: 81 75 751 4169; mshinoha{at}virus.kyoto-u.ac.jp

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