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Related Experiment Videos

Follitropin receptor down-regulation involves a cAMP-dependent post-transcriptional decrease of receptor mRNA

A P Themmen1, L J Blok, M Post

  • 1Department of Endocrinology and Reproduction, Medical Faculty, Erasmus University Rotterdam, The Netherlands.

Molecular and Cellular Endocrinology
|July 1, 1991
PubMed
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Follicle-stimulating hormone (FSH) down-regulates its own receptor mRNA in rat Sertoli cells via a cAMP-mediated, post-transcriptional mechanism. This regulation complements ligand-induced receptor loss, impacting FSH receptor expression.

Area of Science:

  • Reproductive Endocrinology
  • Molecular Endocrinology
  • Cell Biology

Background:

  • Follicle-stimulating hormone (FSH) is crucial for male and female reproduction.
  • FSH exerts its effects by binding to the FSH receptor (FSHR) on target cells, such as Sertoli cells in males.
  • Understanding the regulation of FSHR expression is vital for comprehending reproductive processes.

Purpose of the Study:

  • To investigate the mechanisms by which FSH regulates its own receptor mRNA and protein levels in cultured rat Sertoli cells.
  • To elucidate the role of cyclic AMP (cAMP) and protein synthesis in FSH-induced FSHR downregulation.
  • To differentiate between transcriptional and post-transcriptional regulation of FSHR gene expression.

Main Methods:

  • Primary culture of Sertoli cells isolated from 21-day-old rats.

Related Experiment Videos

  • Treatment with FSH, dibutyryl cyclic AMP (dbcAMP), forskolin, methyl-isobutylxanthine (MIX), and cycloheximide.
  • Quantification of FSH receptor mRNA levels using quantitative assays.
  • Measurement of FSH binding to cell surface receptors using radiolabeled FSH (125I-FSH).
  • Transcriptional run-on assays to assess gene transcription initiation.

    • Main Results:

    • FSH induced a rapid, dose-dependent downregulation of FSH receptor mRNA, which returned to baseline by 16 hours.
    • The effects of FSH on receptor mRNA were mimicked by dbcAMP and forskolin, and prolonged by MIX, indicating cAMP mediation.
    • FSH and dbcAMP reduced FSH receptor mRNA even in the presence of cycloheximide, suggesting a direct, post-transcriptional effect.
    • Transcriptional run-on assays showed FSH did not inhibit FSH receptor gene initiation.
    • FSH treatment rapidly decreased FSH binding (ligand-induced sequestration), while dbcAMP caused a loss of binding sites after 24 hours, likely due to reduced mRNA.

    Conclusions:

    • FSH receptor downregulation in Sertoli cells involves both ligand-induced sequestration and a novel cAMP-mediated, post-transcriptional downregulation of FSH receptor mRNA.
    • The cAMP signaling pathway plays a critical role in modulating FSH receptor expression at the post-transcriptional level.
    • These findings provide a more comprehensive understanding of the autoregulation of the FSH/FSHR system in Sertoli cells.