Capillary morphogenesis gene (CMG)‐1 is among the genes differentially expressed in mouse male germ line stem cells and embryonic stem cells
Capillary morphogenesis gene (CMG)‐1 is among the genes differentially expressed in mouse male germ line stem cells and embryonic stem cells
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DOI：
10.1002/mrd.20504
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发表时间：
2006-08
影响因子：
2.5
通讯作者：
Ryu-Suke Fujino;Y. Ishikawa;Kiyoko Tanaka;Mito Kanatsu-Shinohara;K. Tamura;H. Kogo;T. Shinohara
中科院分区：
文献类型：
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作者：
Ryu-Suke Fujino;Y. Ishikawa;Kiyoko Tanaka;Mito Kanatsu-Shinohara;K. Tamura;H. Kogo;T. Shinohara
我们最近建立了一种在长期培养中扩增雄性生殖干细胞（GS）而不丧失其生精能力的技术。为了深入了解这些细胞的遗传程序，我们使用DNA微阵列比较了GS细胞和胚胎干细胞（ES）的mRNA表达谱。与ES细胞相比，我们发现了79个在GS细胞中上调的基因，包括联会复合物蛋白-1，在无精子症样中缺失，泛素结合酶E2 B和泛素羧基末端水解酶L1，所有这些都对精子发生具有重要的功能。此外，我们鉴定了编码毛细血管形态发生基因（CMG）-1的小鼠直系同源物的cDNA。CMG-1转录本主要在小鼠睾丸的精原细胞和精母细胞中产生。在小鼠精母细胞来源的细胞系GC-2spd（ts）中，通过靶向特异性短干扰RNA减弱CMG-1表达时，细胞形态发生变化，细胞周期蛋白D2的表达消失。使用小鼠细胞周期蛋白D2基因上游的基因组区域的报告基因测定揭示，这种下调发生在转录水平。我们在转染的COS 7细胞的细胞核中检测到FLAG标记的CMG-1蛋白，这表明CMG-1可能在细胞周期蛋白D2基因的转录调控中发挥独特的作用。本研究中发现的GS基因表达上调，将为今后精原干细胞的研究和男性生殖细胞分化的早期阶段提供有用的信息。摩尔Reprod. Dev. 955-966，2006。© 2006 Wiley利斯公司
We recently established a technique to expand male germ line stem (GS) cells in long‐term culture without losing their spermatogenic capacity. To gain insight into the genetic program of these cells, we compared the mRNA expression profile of GS cells with that of embryonic stem (ES) cells using DNA microarrays. We found 79 genes that were upregulated in GS cells compared to ES cells, including synaptonemal complex protein‐1, deleted in azoospermia‐like, ubiquitin‐conjugating enzyme E2B, and ubiquitin carboxy‐terminal hydrolase L1, all of which are functionally important for spermatogenesis. In addition, we identified a cDNA encoding the mouse ortholog of capillary morphogenesis gene (CMG)‐1. CMG‐1 transcripts were predominantly produced in spermatogonia and spermatocytes in mouse testis. When CMG‐1 expression was attenuated in a mouse spermatocyte‐derived cell line, GC‐2spd(ts), by a target‐specific short interfering RNA, the morphology of the cells was changed and the expression of cyclin D2 was abrogated. A reporter assay using a genomic region upstream of the mouse cyclin D2 gene revealed that this downmodulation occurs at the transcriptional level. We detected FLAG‐tagged CMG‐1 protein in the nuclei of transfected COS7 cells, suggesting that CMG‐1 may play a unique role in the transcriptional regulation of the cyclin D2 gene. The upregulated GS genes identified in this study will provide useful information for the future investigation of spermatogonial stem cells and the early phase of male germ cell differentiation. Mol. Reprod. Dev. 955–966, 2006. © 2006 Wiley‐Liss, Inc.