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CDK5RAP3 Regulates Testosterone Production in Mouse Leydig Cells.

Jian Ruan1,2, Qianyi Dong1, Yufan Jin1

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International Journal of Molecular Sciences
|January 28, 2026
PubMed
Summary

Cyclin-dependent kinase 5 regulatory subunit-associated protein 3 (CDK5RAP3) is crucial for testosterone production in Leydig cells. Reduced CDK5RAP3 levels are linked to male infertility and impaired steroidogenesis.

Keywords:
CDK5RAP3CEBPBLeydig cellsSMAD4testosterone

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Area of Science:

  • Endocrinology
  • Reproductive Biology
  • Molecular Biology

Background:

  • Testosterone (T) production by Leydig cells (LCs) is vital for male reproduction, but its regulation is not fully understood.
  • Cyclin-dependent kinase 5 regulatory subunit-associated protein 3 (CDK5RAP3) was identified as a protein involved in steroidogenesis and cholesterol metabolism.

Purpose of the Study:

  • To investigate the role of CDK5RAP3 in Leydig cell development and steroidogenesis.
  • To explore the potential link between CDK5RAP3 expression and male infertility.

Main Methods:

  • Immunoprecipitation-mass spectrometry (IP-MS) in mouse testicular tissue.
  • Analysis of CDK5RAP3 expression in human samples and an ethane dimethanesulfonate (EDS)-induced rat model.
  • Functional studies in primary LCs and MLTC-1 cells, including CDK5RAP3 knockdown and overexpression.
  • In vivo studies using AAV2/9-mediated CDK5RAP3 silencing in adult mice.
  • Mechanistic studies involving protein-protein interactions and pathway analysis (BMP signaling).

Main Results:

  • CDK5RAP3 expression was reduced in LCs of patients with spermatogenic failure and increased during Leydig cell maturation and regeneration.
  • CDK5RAP3 nuclear translocation occurred upon hCG stimulation; its knockdown impaired testosterone production and reduced steroidogenic gene expression (STAR, CYP11A1, CYP17A1, HSD3B).
  • CDK5RAP3 overexpression enhanced testosterone production; in vivo silencing significantly reduced serum testosterone levels.
  • CDK5RAP3 interacted with SMAD4 and CEBPB, and BMP pathway inhibition rescued testosterone deficits.

Conclusions:

  • CDK5RAP3 is an essential regulator of Leydig cell steroidogenesis.
  • CDK5RAP3 plays a significant role in maintaining testosterone levels and may be relevant to male infertility associated with low testosterone.