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Updated: Oct 4, 2026

Isolation of Sertoli Cells and Peritubular Cells from Rat Testes
11:11

Isolation of Sertoli Cells and Peritubular Cells from Rat Testes

Published on: February 8, 2016

Characterization of muscarinic acetylcholine receptor in rat Sertoli cells

M O Borges1, M L Abreu, C S Porto

  • 1Section of Experimental Endocrinology (M.L.C.A., C.S.P., M.C.W.A.), Department of Pharmacology, Universidade Federal de São Paulo-Escola Paulista de Medicina, São Paulo, Brazil 04044-020.

Endocrinology
|October 19, 2001
PubMed

Insights

Muscarinic acetylcholine receptors (mAChRs) are present in rat Sertoli cells at both mRNA and protein levels. mAChR activation influences intracellular signaling pathways, including cAMP accumulation and phosphoinositide hydrolysis.

Area of Science:

  • Reproductive Biology
  • Cellular Signaling
  • Neuroendocrinology

Background:

  • Sertoli cells play a crucial role in spermatogenesis.
  • Muscarinic acetylcholine receptors (mAChRs) are implicated in various physiological processes.
  • The presence and function of mAChRs in Sertoli cells remain largely uncharacterized.

Purpose of the Study:

  • To investigate the presence and subtypes of mAChRs in primary cultured rat Sertoli cells.
  • To elucidate the functional roles of mAChRs in Sertoli cell signaling pathways.
  • To determine the impact of mAChR activation on intracellular signaling and gene expression.

Main Methods:

  • Reverse transcription-polymerase chain reaction (RT-PCR) to detect mAChR mRNA.
  • Ribonuclease protection assay to confirm mRNA transcripts.
  • Radioligand binding studies using [(3)H]quinuclidinyl benzilate.
  • Analysis of intracellular signaling pathways (cAMP, inositol phosphates) upon carbachol stimulation.
  • Gel shift assays to assess transcription factor activation.

Main Results:

  • RT-PCR and ribonuclease protection assay confirmed the expression of m1-m4 mAChR mRNA subtypes.
  • Radioligand binding revealed a single class of mAChR binding sites.
  • Carbachol inhibited forskolin-induced cAMP accumulation and increased inositol phosphates production.
  • These effects were differentially antagonized by specific mAChR blockers.
  • Carbachol stimulated activator protein-1 DNA-binding activity.

Conclusions:

  • Rat Sertoli cells express functional mAChRs at both mRNA and protein levels.
  • mAChR activation modulates adenylyl cyclase activity and phosphoinositide hydrolysis in Sertoli cells.
  • mAChR signaling in Sertoli cells may regulate gene expression, suggesting a role in reproductive function.

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