Transcription profiling of testis from Sertoli cell-selective androgen receptor knockout mice and wild type mice to unravel the molecular mechanisms mediating the effects of androgens on spermatogenesis

To unravel the molecular mechanisms mediating the effects of androgens on spermatogenesis, testicular gene expression was compared in mice with a Sertoli cell-selective androgen receptor knockout (SCARKO) and littermate controls on postnatal d 10. At this age testicular cell composition is still comparable in SCARKOs and controls. Microarray analysis identified 692 genes with significant differences in expression. A more than 2-fold up- or downregulation by androgen action in Sertoli cells was observed for 28 and 6 genes respectively. The biological relevance of the ""strongly"" upregulated genes was supported by the finding that several of them were previously described to be androgen-regulated or essential for spermatogenesis. Serine protease inhibitors were overrepresented in the same subgroup suggesting a role for androgens in cell junction dynamics and tissue restructuring events during spermatogenesis. A time course experiment (d8-d20), followed by cluster analysis allowed the identification of typical expression patterns of differentially expressed testicular genes during initiation of spermatogenesis. Three genes with a pattern closely resembling that of Pem, a prototypal androgen-regulated gene in Sertoli cells, were selected for confirmation by RT-PCR and further analysis. The data confirm that the SCARKO model allows identification of novel androgen-regulated genes in the testis. This particular series represents all data from d 10. The additional expression data from the time course (d8-d20) is represented by series GSE2259 (Testicular gene expression in SCARKO mice during prepuberty). Keywords = SCARKO Keywords = testis Keywords = spermatogenesis Keywords = androgens Keywords = microarray Keywords = Pem Keywords = serine protease inhibitor为阐明介导雄激素对精子发生作用的分子机制，本研究对出生后第10天的塞尔托利细胞（Sertoli cell）选择性雄激素受体敲除（SCARKO）小鼠及其同窝对照小鼠的睾丸基因表达水平进行了比较分析。该时间点下，SCARKO小鼠与对照小鼠的睾丸细胞组成仍无显著差异。芯片（microarray）分析共鉴定出692个表达存在显著差异的基因，其中在塞尔托利细胞中受雄激素调控出现2倍以上上调和下调的基因分别为28个和6个。部分"显著上调"的基因此前已被报道为雄激素调控基因或精子发生必需基因，这佐证了本数据集的生物学相关性。该上调基因亚组中丝氨酸蛋白酶抑制剂（serine protease inhibitor）的富集度显著升高，提示雄激素在精子发生过程中参与细胞连接动态调控与组织重构事件。通过对出生后第8天至第20天的小鼠开展时间梯度实验，并结合聚类分析，我们鉴定出精子发生起始阶段睾丸差异表达基因的典型表达谱。选取3个表达谱与塞尔托利细胞中典型雄激素调控基因Pem高度相似的基因，通过逆转录聚合酶链反应（RT-PCR）进行表达验证与后续分析。本研究数据证实，SCARKO模型可用于鉴定睾丸中新型雄激素调控基因。本数据集涵盖出生后第10天的全部实验数据，而出生后第8天至第20天的时间梯度表达数据则收录于数据集GSE2259（《青春期前SCARKO小鼠睾丸基因表达分析》）。关键词：塞尔托利细胞选择性雄激素受体敲除（SCARKO）、睾丸、精子发生、雄激素、芯片（microarray）、Pem、丝氨酸蛋白酶抑制剂（serine protease inhibitor）