Identification of testosterone-/androgen receptor-regulated genes in mouse Sertoli cells.
Identification of testosterone-/androgen receptor-regulated genes in mouse Sertoli cells.
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DOI：
10.1038/aja.2011.94
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发表时间：
2012-03
影响因子：
2.9
通讯作者：
Qiaoxia Zhang;Xiao-Yan Zhang;Zhen-Ming Zhang;Wei Lu;Ling Liu;Gang Li;Z. Cai;Y. Gui;Chawnshang Chang
中科院分区：
文献类型：
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作者：
Qiaoxia Zhang;Xiao-Yan Zhang;Zhen-Ming Zhang;Wei Lu;Ling Liu;Gang Li;Z. Cai;Y. Gui;Chawnshang Chang
雄激素和雄激素受体 (AR) 在男性精子发生和生育能力中发挥着重要作用，但支持细胞中详细的雄激素/AR 信号仍不清楚。为了鉴定支持细胞中的 AR 靶基因，我们分析了支持细胞中缺乏 AR 的小鼠 (S-AR−/y) 及其同窝野生型 (WT) 小鼠的睾丸基因表达谱。数字基因表达分析发现，与 WT 小鼠相比，S-AR−/y 小鼠睾丸中有 2276 个基因下调，2865 个基因上调。为了进一步确定支持细胞内的差异，我们首先构建了稳定转染AR的支持细胞系TM4（命名为TM4/AR），并发现雄激素未能反式激活支持细胞中的AR。有趣的是，额外瞬时转染 AR-cDNA 导致 TM4/AR 细胞产生显着的雄激素反应，其雄激素敏感性比 TM4 细胞高 10 倍。在表现出最大雄激素反应的条件下，我们分析了基因表达，发现在睾酮存在下瞬时转染 AR-cDNA 使 2313 个基因的表达水平改变了两倍以上。在这些基因中，在 S-AR−/y 小鼠睾丸和 TM4/AR 细胞中发现了 603 个雄激素/AR 调节基因，其中 164 个上调和 439 个下调。利用信息学分析，应用基因本体论分析这些雄激素/AR调节基因，以预测雄激素/AR在精子发生过程中的潜在作用。总之，在 S-AR−/y 小鼠睾丸和 TM4/AR 细胞中进行基因分析可能有助于我们更好地了解支持细胞中的雄激素/AR 信号及其对精子发生的影响。
Androgen and androgen receptor (AR) play important roles in male spermatogenesis and fertility, yet detailed androgen/AR signals in Sertoli cells remain unclear. To identify AR target genes in Sertoli cells, we analyzed the gene expression profiles of testis between mice lacking AR in Sertoli cells (S-AR−/y) and their littermate wild-type (WT) mice. Digital gene expression analysis identified 2276 genes downregulated and 2865 genes upregulated in the S-AR−/y mice testis compared to WT ones. To further nail down the difference within Sertoli cells, we first constructed Sertoli cell line TM4 with stably transfected AR (named as TM4/AR) and found androgens failed to transactivate AR in Sertoli TM4 and TM4/AR cells. Interestingly, additional transient transfection of AR-cDNA resulted in significant androgen responsiveness with TM4/AR cells showing 10 times more androgen sensitivity than TM4 cells. In the condition where maximal androgen response was demonstrated, we then analyzed gene expression and found the expression levels of 2313 genes were changed more than twofold by transient transfection of AR-cDNA in the presence of testosterone. Among these genes, 603 androgen-/AR-regulated genes, including 164 upregulated and 439 downregulated, were found in both S-AR−/y mice testis and TM4/AR cells. Using informatics analysis, the gene ontology was applied to analyze these androgen-/AR-regulated genes to predict the potential roles of androgen/AR in the process of spermatogenesis. Together, using gene analysis in both S-AR−/y mice testis and TM4/AR cells may help us to better understand the androgen/AR signals in Sertoli cells and their influences in spermatogenesis.