Presumptive pre-Sertoli cells express genes involved in cell proliferation and cell signalling during a critical window in early testis differentiation
Presumptive pre-Sertoli cells express genes involved in cell proliferation and cell signalling during a critical window in early testis differentiation
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DOI：
10.1002/mrd.20722
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发表时间：
2007-12-01
影响因子：
2.5
通讯作者：
Silversides, David W.
中科院分区：
文献类型：
--
作者：
Cory, Aron T.;Boyer, Alexandre;Silversides, David W.
在哺乳动物中，雄性生殖嵴的前支持细胞是第一个显示性别特异性分化和差异基因表达的细胞类型。驱动前支持细胞分化和最终睾丸形成的遗传级联反应开始被解开，但仍存在许多问题。更好地了解前支持细胞的转录组后立即性别决定是必不可少的，以进一步了解这一分化过程。使用在杂交小鼠/猪SRY启动子（HybSRYp-RFP）控制下表达红色荧光蛋白（RFP）的小鼠模型来纯化来自胚胎第12.0天（e12.0）雄性生殖嵴的细胞。为了比较HybSRYp-RFP细胞群体与年龄匹配的整个雌性生殖嵴的转录组，提取RNA并用于产生分子探针，所述分子探针杂交到Affyoung小鼠基因组430 2.0微阵列上。在表达HybSRYp-RFP的细胞群中鉴定了被认为是前支持细胞的标志物的基因的表达，包括Sox 9、Mis、Dhh和Fgf 9，而未鉴定生殖细胞（Oct 4、SSEA-1）和内皮细胞（Ntrk 3）的标志物。相反，卵巢体细胞表达的标志物，包括Fst和8 mp2，被鉴定为在卵巢细胞群中过表达。在一般情况下，鉴定为在表达HybSRYp-RFP的细胞中过表达2.5倍的基因显著编码细胞信号传导和细胞外蛋白。Sox 10，Stc 2，Fgf 18，Fgf 13和Wnt 6的表达进一步通过全坐骑原位杂交（WISH）在e11.5和e14.5之间的男性和女性生殖嵴上进行了表征。Sox 10、Fgf 18、Fgf 13和Stc 2基因在雄性生殖嵴中均有表达，而Wnt 6基因在雄性和雌性生殖嵴中均有广泛表达。这些数据代表了迄今为止最早的纯化推定前支持细胞的全面微阵列表达分析。
In mammals, the pre-Sertoli cell of the male genital ridge is the first cell type to display sex specific differentiation and differential gene expression. The genetic cascade driving the differentiation of pre-Sertoli cells and ultimately testis formationis beginning to be unravelled, but many questions remain. A better understanding of the transcriptome of pre-Sertoli cells immediately after sex determination is essential in order to further understand this differentiation process. A mouse model expressing Red Fluorescent Protein (RFP) under the control of a hybrid mouse/pig SRY promoter (HybSRYp-RFP) was used to purify cells from embryonic day 12.0 (e12.0) male genital ridges. To compare the transcriptomes of HybSRYp-RFP cell populations versus age matched whole female genital ridges, RNA was extracted and used to generate molecular probes that were hybridized onto Affymetrix Mouse Genome 430 2.0 microarrays. The expression of genes considered markers for pre-Sertoli cells, including Sox9, Mis, Dhh and Fgf9 were identified within the HybSRYp-RFP expressing cell population, while markers for germ cells (Oct4, SSEA-1) and endothelial cells (Ntrk3) were not identified. In contrast, markers for ovarian somatic cell expression, including Fst and 8mp2, were identified as overexpressed within the ovarian cell population. In a general fashion, genes identified as 2.5-fold over expressed in HybSRYp-RFP expressing cells coded notably for cell signalling and extra cellular proteins. The expression of Sox10, Stc2, Fgf18, Fgf13 and Wnt6 were further characterized via whole mount in situ hybridization (WISH) on male and female genital ridges between e11.5 and e14.5. Sox10, Fgf18, Fgf13 and Stc2 gene expression was detected within the male genital ridges while Wnt6 was found diffusely within both the male and female genital ridges. These data represent the earliest comprehensive microarray expression analysis of purified presumptive pre-Sertoli cells available to date.