Jiang Keer, Fu Zirui, Tsourkas Philippos, Kothandapani Anbarasi, Kearse Tyler, McIlwain Sean J, Mayère Chloé, Nef Serge, Jorgensen Joan S
Department of Comparative Biosciences, School of Veterinary Medicine, University of Wisconsin, Madison, WI 53706.
Cellular and Molecular Biology Graduate Program, University of Wisconsin, Madison, WI 53706.
Proc Natl Acad Sci U S A. 2025 Jun 10;122(23):e2501392122. doi: 10.1073/pnas.2501392122. Epub 2025 Jun 3.
Historically, endocrine cells were perceived to coordinate their output in a uniform manner. Recently however, single-cell technologies have uncovered heterogeneity within these populations, indicating that individual cells may operate as independently regulated units. Using high-resolution tools such as single-molecule fluorescent in situ hybridization (sm-FISH) and single-cell RNA sequencing (scRNA-seq), we investigated the contributions of individual and the collective of fetal Leydig cells to androgen production over time during mouse testis development. Temporal profiles of intratesticular androgens alongside the expression of steroidogenic pathway genes ( and ) from prenatal to perinatal testes demonstrated that the peak in gene expression preceded the peak in androgen production. Spatially, steroidogenic cells were initially observed to be concentrated toward the anterior-posterior poles along the center of the dorsal-ventral axis of the fetal testis at embryonic day (E) 13 and then expanded to a uniform distribution by E16. Next, sm-FISH using probes for individual steroidogenic pathway genes exposed the following findings: gene transcription and processing of individual and combinations of steroidogenic pathway genes are not synchronized among fetal Leydig cells; and some fetal Leydig cells express incomplete sets of genes. Further, sm-FISH and scRNA-seq data corroborated the presence of fetal Leydig and other interstitial cell types harboring incomplete sets of steroidogenic pathway genes throughout developmental stages. Taken together, these findings highlight that fetal steroidogenic gene expression is tightly regulated and that transcript presence among interstitial cell types promotes the possibility that optimal androgen biosynthesis results from a cooperative effort among neighboring steroidogenic cells.
从历史上看，内分泌细胞被认为是以统一的方式协调其输出。然而，最近单细胞技术揭示了这些细胞群体中的异质性，这表明单个细胞可能作为独立调节的单位发挥作用。我们使用单分子荧光原位杂交（sm-FISH）和单细胞RNA测序（scRNA-seq）等高分辨率工具，研究了小鼠睾丸发育过程中胎儿睾丸间质细胞个体及其集体对雄激素产生随时间的贡献。从产前到围产期睾丸，睾丸内雄激素水平以及类固醇生成途径基因（和）表达随时间的变化表明，基因表达峰值先于雄激素产生峰值。在空间上观察到类固醇生成细胞最初在胚胎第13天沿着胎儿睾丸背腹轴中心集中于前后极，然后在胚胎第16天扩展为均匀分布。接下来，使用针对单个类固醇生成途径基因的探针进行sm-FISH得到以下结果：胎儿睾丸间质细胞中类固醇生成途径基因的个体转录和组合转录及加工不同步；并且一些胎儿睾丸间质细胞表达不完整套的基因。此外，sm-FISH和scRNA-seq数据证实了在整个发育阶段都存在具有不完整套类固醇生成途径基因的胎儿睾丸间质细胞和其他间质细胞类型。综上所述，这些发现突出表明胎儿类固醇生成基因表达受到严格调控以及间质细胞类型之间转录本的存在增加了最佳雄激素生物合成是由相邻类固醇生成细胞协同努力的可能性。
