A role for TRPC3 in mammalian testis development

Zhenhua Ming; Stefan Bagheri-Fam; Emily R Frost; Janelle M Ryan; Vincent R Harley

doi:10.3389/fcell.2024.1337714

A role for TRPC3 in mammalian testis development

Front Cell Dev Biol. 2024 Feb 15:12:1337714. doi: 10.3389/fcell.2024.1337714. eCollection 2024.

Authors

Zhenhua Ming^{1

2}, Stefan Bagheri-Fam¹, Emily R Frost¹, Janelle M Ryan^{1

2}, Vincent R Harley^{1

2}

Affiliations

¹ Sex Development Laboratory, Hudson Institute of Medical Research, Melbourne, VIC, Australia.
² Department of Molecular and Translational Science, Monash University, Melbourne, VIC, Australia.

Abstract

SOX9 is a key transcription factor for testis determination and development. Mutations in and around the SOX9 gene contribute to Differences/Disorders of Sex Development (DSD). However, a substantial proportion of DSD patients lack a definitive genetic diagnosis. SOX9 target genes are potentially DSD-causative genes, yet only a limited subset of these genes has been investigated during testis development. We hypothesize that SOX9 target genes play an integral role in testis development and could potentially be causative genes in DSD. In this study, we describe a novel testicular target gene of SOX9, Trpc3. Trpc3 exhibits high expression levels in the SOX9-expressing male Sertoli cells compared to female granulosa cells in mouse fetal gonads between embryonic day 11.5 (E11.5) and E13.5. In XY Sox9 knockout gonads, Trpc3 expression is markedly downregulated. Moreover, culture of E11.5 XY mouse gonads with TRPC3 inhibitor Pyr3 resulted in decreased germ cell numbers caused by reduced germ cell proliferation. Trpc3 is also expressed in endothelial cells and Pyr3-treated E11.5 XY mouse gonads showed a loss of the coelomic blood vessel due to increased apoptosis of endothelial cells. In the human testicular cell line NT2/D1, TRPC3 promotes cell proliferation and controls cell morphology, as observed by xCELLigence and HoloMonitor real-time analysis. In summary, our study suggests that SOX9 positively regulates Trpc3 in mouse testes and TRPC3 may mediate SOX9 function during Sertoli, germ and endothelial cell development.

Keywords: DSD; SOX9; TRP; TRPC3; sertoli cells; sex determination; testis.

Grants and funding

The author(s) declare financial support was received for the research, authorship, and/or publication of this article. This research received financial support from the National Health and Medical Research Council Program Grant 2002426 and Fellowship APP1154870 awarded to VH. Additional support was provided by the China Scholarship Council (CSC) and the Australian Government Research Training Program Scholarship for ZM. We also acknowledge funding through the Victorian Government’s Operational Infrastructure Support Program.