New candidate gene identification for controlling mammalian gonadal sex determination. Mus musculus

Mammalian gonadal sex determination is dependent on proper expression of sex determining genes in fetal gonadal somatic support cells (i.e., pre-granulosa and pre-Sertoli cells in XX and XY gonads, resp.). We used a unique transgenic mouse strain combined with microarray profiling to identify all the differentially expressed transcripts in XX and XY isolated somatic support cells during critical stages of gonadal development and differentiation. Overall design: XX and XY somatic support cells (SSC) were isolated by flow cytometry from embryonic day (E) 11.5 and E12.5 mouse gonads. Total RNA was isolated from pools of isolated cells; 3 pools per sex and each timepoint.

哺乳动物性腺性别决定依赖于胎儿性腺体细胞支持细胞内性别决定基因的精准表达（即XX与XY性腺中分别对应的前颗粒细胞与前支持细胞）。本研究借助独特的转基因小鼠品系结合微阵列（microarray）表达谱分析，在性腺发育与分化的关键阶段，对分离获取的XX与XY型体细胞支持细胞（SSC）中的全部差异表达转录本进行了系统鉴定。实验设计概述：通过流式细胞术，从胚胎第11.5天（E11.5）与胚胎第12.5天（E12.5）的小鼠性腺中分离XX与XY型体细胞支持细胞；从收集的细胞混合样本中提取总RNA，每个性别及每个时间点均设置3个生物学重复混合样本。