Genetic reconstruction of mouse spermatogonial stem cell self-renewal in vitro by Ras-cyclin D2 activation.
Spermatogonial stem cells (SSCs) undergo self-renewal division and support spermatogenesis. Although several cytokines coordinate to drive SSC self-renewal, little is known about the mechanisms underlying this process. We investigated the molecular mechanism by reconstructing SSC self-renewal in vitro without exogenous cytokines. Activation of Ras or overexpression of cyclins D2 and E1, both of which were induced by Ras, enabled long-term self-renewal of cultured spermatogonia. SSCs with activated Ras responded properly to differentiation signals and underwent spermatogenesis, whereas differentiation was abrogated in cyclin transfectants after spermatogonial transplantation. Both Ras- and cyclin-transfected cells produced seminomatous tumors, suggesting that excessive self-renewing stimulus induces oncogenic transformation. In contrast, cells that overexpressed cyclin D1 or D3 failed to make germ cell colonies after transplantation, which indicated that cyclin expression pattern is an important determinant to long-term SSC recolonization. Thus, the Ras-cyclin D2 pathway regulates the balance between tissue maintenance and tumorigenesis in the SSC population.
Lee
Jiyoung
J
Department of Molecular Genetics, Graduate School of Medicine, Kyoto University, Kyoto 606-8501, Japan.
Kanatsu-Shinohara
Mito
M
Morimoto
Hiroko
H
Kazuki
Yasuhiro
Y
Takashima
Seiji
S
Oshimura
Mitsuo
M
Toyokuni
Shinya
S
Shinohara
Takashi
T
D000818:Animals; D002453:Cell Cycle; D002478:Cells, Cultured; D056742:Cyclin D2; D016213:Cyclins / Q000235:genetics* / Q000378:metabolism; D016207:Cytokines / Q000378:metabolism; D011905:Genes, ras / Q000502:physiology; D008297:Male; D051379:Mice; D013093:Spermatogonia / Q000166:cytology* / Q000254:growth & development; D033581:Stem Cell Transplantation; D013234:Stem Cells / Q000166:cytology* / Q000378:metabolism; D013736:Testicular Neoplasms / Q000235:genetics; D013737:Testis / Q000166:cytology / Q000378:metabolism; D014162:Transfection; D018631:ras Proteins / Q000235:genetics* / Q000378:metabolism
C486196:Ccnd2 protein, mouse; D056742:Cyclin D2; D016213:Cyclins; D016207:Cytokines; D018631:ras Proteins