McClelland Kathryn S, Bell Katrina, Larney Christian, Harley Vincent R, Sinclair Andrew H, Oshlack Alicia, Koopman Peter, Bowles Josephine
Institute for Molecular Bioscience, The University of Queensland, Brisbane, Queensland, Australia.
Murdoch Childrens Research Institute, Royal Children's Hospital, Melbourne, Victoria, Australia.
Biol Reprod. 2015 Jun;92(6):145. doi: 10.1095/biolreprod.115.128918. Epub 2015 Apr 8.
Male sex determination hinges on the development of testes in the embryo, beginning with the differentiation of Sertoli cells under the influence of the Y-linked gene SRY. Sertoli cells then orchestrate fetal testis formation including the specification of fetal Leydig cells (FLCs) that produce steroid hormones to direct virilization of the XY embryo. As the majority of XY disorders of sex development (DSDs) remain unexplained at the molecular genetic level, we reasoned that genes involved in FLC development might represent an unappreciated source of candidate XY DSD genes. To identify these genes, and to gain a more detailed understanding of the regulatory networks underpinning the specification and differentiation of the FLC population, we developed methods for isolating fetal Sertoli, Leydig, and interstitial cell-enriched subpopulations using an Sf1-eGFP transgenic mouse line. RNA sequencing followed by rigorous bioinformatic filtering identified 84 genes upregulated in FLCs, 704 genes upregulated in nonsteroidogenic interstitial cells, and 1217 genes upregulated in the Sertoli cells at 12.5 days postcoitum. The analysis revealed a trend for expression of components of neuroactive ligand interactions in FLCs and Sertoli cells and identified factors potentially involved in signaling between the Sertoli cells, FLCs, and interstitial cells. We identified 61 genes that were not known previously to be involved in specification or differentiation of FLCs. This dataset provides a platform for exploring the biology of FLCs and understanding the role of these cells in testicular development. In addition, it provides a basis for targeted studies designed to identify causes of idiopathic XY DSD.
男性性别决定取决于胚胎中睾丸的发育，始于在Y连锁基因SRY影响下支持细胞的分化。支持细胞随后协调胎儿睾丸的形成，包括产生类固醇激素以指导XY胚胎男性化的胎儿睾丸间质细胞（FLC）的特化。由于大多数XY性发育障碍（DSD）在分子遗传水平上仍无法解释，我们推测参与FLC发育的基因可能是未被重视的XY DSD候选基因来源。为了鉴定这些基因，并更详细地了解支撑FLC群体特化和分化的调控网络，我们利用Sf1-eGFP转基因小鼠品系开发了分离胎儿支持细胞、睾丸间质细胞和富含间质细胞亚群的方法。RNA测序随后经过严格的生物信息学筛选，确定了在妊娠12.5天时FLC中上调的84个基因、非类固醇生成间质细胞中上调的704个基因以及支持细胞中上调的1217个基因。分析揭示了神经活性配体相互作用成分在FLC和支持细胞中表达的一种趋势，并确定了可能参与支持细胞、FLC和间质细胞之间信号传导的因子。我们鉴定出61个以前未知参与FLC特化或分化的基因。该数据集为探索FLC生物学特性和了解这些细胞在睾丸发育中的作用提供了一个平台。此外，它为旨在确定特发性XY DSD病因的靶向研究提供了基础。
