Divergent regulatory element programs steer sex-specific supporting cell differentiation along mouse gonadal development [RNA-Seq]

Identifying cis-regulatory elements is essential for understanding how gene expression is precisely controlled during critical biological processes like sex determination. In this study, we aimed to uncover the cis-regulatory regions involved in the differentiation of supporting cells as Sertoli cells or pre-granulosa cells in the mouse gonads. To achieve this, we combined RNA sequencing (RNA-seq) and Assay for Transposase-Accessible Chromatin sequencing (ATAC-seq) of purified supporting cells at four developmental stages in both sexes. RNA-seq enabled us to capture sexual and temporal changes in gene expression during cell differentiation, while ATAC-seq provided insight into chromatin accessibility, pinpointing regulatory regions that control these expression patterns. By integrating these two datasets, we identified specific cis-regulatory elements that likely guide the fate of supporting cells. Our findings shed light on the complex gene regulatory networks involved in sex fate differentiation, highlighting the critical role of cis-regulatory elements in orchestrating this process. These insights may also have implications for understanding disorders of sexual development (DSDs). We purified by FACS pre-granulosa and Sertoli cells from mouse fetal gonads and performed bulk RNA-seq and ATAC-seq in triplicates at four developmental stages (E11.5, E12.5, E13.5 and E15.5).顺式调控元件（cis-regulatory elements）的鉴定，对于理解性别决定等关键生物学过程中基因表达的精准调控机制至关重要。本研究旨在揭示小鼠性腺中支持细胞分化为塞尔托利细胞（Sertoli cells）或前颗粒细胞（pre-granulosa cells）过程所涉及的顺式调控区域。为此，我们针对雌雄两性四个发育阶段的纯化支持细胞，结合运用了RNA测序（RNA-seq）与转座酶可及性染色质测序（ATAC-seq）技术。RNA-seq可捕捉细胞分化过程中基因表达的性别特异性与时序性变化，而ATAC-seq则能够解析染色质可及性，精准定位调控此类表达模式的基因组区域。通过整合这两组测序数据集，我们鉴定出了可能指导支持细胞命运决定的特异性顺式调控元件。本研究结果阐明了参与性别命运分化的复杂基因调控网络，并凸显了顺式调控元件在调控该过程中的关键作用。这些发现也可为理解性发育异常（DSDs）提供理论参考。本研究通过荧光激活细胞分选（FACS）从小鼠胎儿性腺中分离前颗粒细胞与塞尔托利细胞，并在四个发育阶段（E11.5、E12.5、E13.5及E15.5）开展了三次生物学重复的批量RNA测序（bulk RNA-seq）与ATAC-seq实验。