Characterization of muscarinic acetylcholine receptor in rat Sertoli cells

Endocrinology
M O BorgesMaria Christina W Avellar

Abstract

This study was designed to characterize muscarinic acetylcholine receptors (mAChRs) in primary cultured Sertoli cells from 30-d-old rats. RT-PCR was performed, and five PCR products corresponding to m1-m5 mAChR mRNA subtypes were detected in these cells. Ribonuclease protection assay further confirmed the presence of protected products for m1, m2, m3, and m4 mAChR transcripts. Radioligand binding studies and the analysis of changes in intracellular signaling pathways after cell exposure to carbachol were performed to study mAChRs at the protein level. Scatchard analysis revealed one single class of [(3)H]quinuclidinyl benzilate binding sites. Carbachol produced a reduction on forskolin-induced intracellular cAMP accumulation in Sertoli cells. This effect was reversed by atropine, methoctramine, and tropicamide but not by p-fluoro-hexahydro-sila-difenidol or pirenzepine. Carbachol also induced an increase on total [(3)H]-inositol phosphates content, an effect antagonized by atropine, p-fluoro-hexahydro-sila-difenidol, or pirenzepine but not by methoctramine. Thus, mAChR activation in Sertoli cell is linked to both adenylyl cyclase inhibition and to phosphoinositide hydrolysis. Furthermore, gel shift assays indicated that carbach...Continue Reading

Citations

Sep 11, 2009·American Journal of Physiology. Regulatory, Integrative and Comparative Physiology·Maureen BashaSamuel Chacko
Aug 21, 2009·Journal of Molecular Neuroscience : MN·Maria Christina W AvellarCatarina S Porto
Sep 2, 2009·Anais Da Academia Brasileira De Ciências·Maria Christina W AvellarCatarina S Porto
Jan 24, 2002·Biology of Reproduction·Daniel B C QueirózMaria Christina W Avellar
Sep 10, 2010·Biology of Reproduction·Gisele Renata Oliveira GomesCatarina Segreti Porto
Feb 13, 2014·International Journal of Developmental Neuroscience : the Official Journal of the International Society for Developmental Neuroscience·Lalit P ChandravanshiVinay K Khanna
May 23, 2020·Environmental Toxicology and Pharmacology·Ifeoluwa O AwogbindinEbenezer O Farombi

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