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Related Experiment Video

Updated: Jul 1, 2025

Expression and Purification of the Human Lipid-sensitive Cation Channel TRPC3 for Structural Determination by Single-particle Cryo-electron Microscopy
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A role for TRPC3 in mammalian testis development.

Zhenhua Ming1,2, Stefan Bagheri-Fam1, Emily R Frost1

  • 1Sex Development Laboratory, Hudson Institute of Medical Research, Melbourne, VIC, Australia.

Frontiers in Cell and Developmental Biology
|March 1, 2024
PubMed
Summary

SOX9 regulates the novel gene Trpc3, crucial for male reproductive development. TRPC3 impacts Sertoli, germ, and endothelial cell functions, potentially explaining some undiagnosed Disorders of Sex Development.

Keywords:
DSDSOX9TRPTRPC3sertoli cellssex determinationtestis

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Area of Science:

  • Reproductive biology
  • Developmental biology
  • Genetics

Background:

  • SOX9 is essential for testis determination and development.
  • Mutations in SOX9 are linked to Disorders of Sex Development (DSD).
  • Many DSD cases lack a genetic diagnosis, suggesting undiscovered causative genes.

Purpose of the Study:

  • To identify novel SOX9 target genes involved in testis development.
  • To investigate the role of the identified gene, Trpc3, in gonadal development.
  • To explore the potential of TRPC3 as a causative gene in DSD.

Main Methods:

  • Gene expression analysis in mouse fetal gonads (E11.5-E13.5).
  • SOX9 knockout mouse models to assess Trpc3 regulation.
  • Pharmacological inhibition of TRPC3 in cultured gonads.
  • Cell proliferation and morphology analysis using xCELLigence and HoloMonitor in human cell lines.

Main Results:

  • Trpc3 is identified as a novel SOX9 target gene with higher expression in male than female fetal gonads.
  • Trpc3 expression is significantly reduced in XY Sox9 knockout gonads.
  • TRPC3 inhibition in XY gonads decreased germ cell proliferation and increased endothelial cell apoptosis, disrupting blood vessel formation.
  • TRPC3 promotes proliferation and influences morphology in human testicular cells.

Conclusions:

  • SOX9 positively regulates Trpc3 expression in developing testes.
  • TRPC3 plays a critical role in Sertoli, germ, and endothelial cell development during testis formation.
  • TRPC3 is a potential mediator of SOX9 function and a candidate gene for undiagnosed DSD.