Characterization of the muscarinic receptor subtype M3 in the bovine zona fasciculata-reticularis cells by receptor binding, mRNA and functional studies
| Autor: | J M Saez, A Janossy, J. Y. Li |
|---|---|
| Přispěvatelé: | Communications Cellulaires et Différenciation (CCD), Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Claude Bernard Lyon 1 (UCBL), Université de Lyon-Université de Lyon-Institut National de la Recherche Agronomique (INRA), ProdInra, Migration |
| Jazyk: | angličtina |
| Rok vydání: | 1999 |
| Předmět: |
medicine.medical_specialty
Carbachol Hydrocortisone Endocrinology Diabetes and Metabolism [SDV]Life Sciences [q-bio] Cell Culture Techniques Gene Expression Biology Endocrinology Zona fasciculata Internal medicine ARN-MESSAGER Muscarinic acetylcholine receptor medicine Cyclic AMP Animals RNA Messenger Receptor Receptor Muscarinic M3 SURRENALE Reverse Transcriptase Polymerase Chain Reaction Muscarinic antagonist Muscarinic acetylcholine receptor M3 Muscarinic acetylcholine receptor M1 Pirenzepine Receptors Muscarinic Zona Reticularis [SDV] Life Sciences [q-bio] medicine.anatomical_structure Cattle Zona Fasciculata medicine.drug |
| Zdroj: | Journal of Endocrinology Journal of Endocrinology, BioScientifica, 1999, 136, pp.329-336 |
| ISSN: | 0022-0795 1479-6805 |
| Popis: | In the present work we have investigated which muscarinic (M) receptor subtype is responsible for the steroidogenic effect of muscarinic agonists in bovine zona fasciculata-reticularis (ZFR) cells in culture. Radioligand binding studies using the muscarinic antagonist [(3)H]quinuclidinyl benzilate ([(3)H]QNB) demonstrated binding sites of high affinity (K(d)=0.45 nM) and low capacity ( approximately 8000 sites/cell). Pharmacological characterisation of muscarinic receptors was assessed by evaluating the effects of the M(3)>M(1)>>M(2) antagonist 4-DAMP (4-diphenylacetyl-N-methylpiperidine) and the M(1)=M(4)> M(3)>>M(2) antagonist pirenzepine on the binding of [(3)H]QNB and carbachol-induced cortisol production. For both parameters, the potency of 4-DAMP was about two orders of magnitude higher than that of pirenzepine. Reverse transcriptase (RT)-PCR analysis of bovine ZFR mRNAs using specific primers for M(2), M(3) and M(4) receptors revealed the expression of only M(3) mRNA. Moreover, carbachol significantly stimulated inositol phosphate accumulation, but had no inhibitory effect on basal or ACTH-induced cAMP production. Indeed, carbachol potentiated ACTH-induced cAMP production and this effect was, in part, mediated through protein kinase C. Lastly, neomycin, an inhibitor of phosphoinositide turnover, significantly attenuated carbachol-evoked cortisol production. Thus, pharmacological, biochemical and mRNA studies indicate that the M(3) receptor subtype is responsible for the biological effects of muscarinic agonists in bovine ZFR cells. |
| Databáze: | OpenAIRE |
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