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 series represents the time course data from d 8, d 10, d 12, d 16 and d 20. The expression data from the additional SCARKO vs control comparison on d 10 is represented by series GSE2260 (Testicular gene expression in SCARKO mice at day 10). Keywords = SCARKO Keywords = testis Keywords = spermatogenesis Keywords = androgens Keywords = microarray Keywords = Pem Keywords = serine protease inhibitor为阐明介导雄激素调控精子发生过程的分子机制，本研究对出生后第10天的支持细胞选择性雄激素受体敲除（Sertoli cell-selective androgen receptor knockout, SCARKO）小鼠与同窝对照小鼠的睾丸基因表达谱开展了比较分析。该日龄下，SCARKO小鼠与对照小鼠的睾丸细胞组成仍较为相似。基因芯片（microarray）分析共鉴定出692个表达存在显著差异的基因。其中，受支持细胞雄激素调控且表达上调、下调幅度超过2倍的基因分别有28个和6个。针对“强上调”基因的生物学相关性验证结果显示，其中多个基因此前已被报道为雄激素调控基因，或是精子发生所必需的基因。丝氨酸蛋白酶抑制剂（serine protease inhibitor）在该上调亚组中显著富集，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。本研究对出生后第8天至第20天的小鼠开展时间进程实验，并通过聚类分析鉴定出精子发生启动阶段差异表达睾丸基因的典型表达模式。我们选取了3个表达模式与支持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式反应（RT-PCR）进行验证并开展后续分析。本研究数据证实，SCARKO模型可用于鉴定睾丸组织中新型的雄激素调控基因。本数据集包含出生后第8、10、12、16及20天的时间进程表达数据。额外的出生后第10天SCARKO小鼠与对照小鼠的比较表达数据，已收录于系列数据集GSE2260《SCARKO小鼠出生第10天的睾丸基因表达》。关键词：SCARKO、睾丸、精子发生、雄激素、基因芯片、Pem、丝氨酸蛋白酶抑制剂