RGD Reference Report - ETA receptor-mediated Ca2+ mobilisation in H9c2 cardiac cells. - Rat Genome Database

RGD Reference Report - ETA receptor-mediated Ca2+ mobilisation in H9c2 cardiac cells. - Rat Genome Database

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ETA receptor-mediated Ca2+ mobilisation in H9c2 cardiac cells.
Authors:	Ceccarelli, F Scavuzzo, MC Giusti, L Bigini, G Costa, B Carnicelli, V Zucchi, R Lucacchini, A Mazzoni, MR
Citation:	Ceccarelli F, etal., Biochem Pharmacol 2003 Mar 1;65(5):783-93.
RGD ID:	728499
Pubmed:	PMID:12628492 (View Abstract at PubMed)
Expression and pharmacological properties of endothelin receptors (ETRs) were investigated in H9c2 cardiomyoblasts. The mechanism of receptor-mediated modulation of intracellular Ca(2+) concentration ([Ca(2+)](i)) was examined by measuring fluorescence increase of Fluo-3-loaded cells with flow cytometry. Binding assays showed that [125I]endothelin-1 (ET-1) bound to a single class of high affinity binding sites in cardiomyoblast membranes. Endothelin-3 (ET-3) displaced bound [125I]ET-1 in a biphasic manner, in contrast to an ET(B)-selective agonist, IRL-1620, that was ineffective. The ET(B)-selective antagonist, BQ-788, inhibited [125I]ET-1 binding in a monophasic manner and with low potency. An ET(A)-selective antagonist, BQ-123, competed [125I]ET-1 binding in a monophasic manner. This antagonist was found to be 13-fold more potent than BQ-788. Immunoblotting analysis using anti-ET(A) and -ET(B) antibodies confirmed a predominant expression of the ET(A) receptor. ET-1 induced a concentration-dependent increase of Fluo-3 fluorescence in cardiomyoblasts resuspended in buffer containing 1mM CaCl(2). Treatment of cells with antagonists, PD-145065 and BQ-123, or a phospholipase C-beta inhibitor, U-73122, abolished ET-1-mediated increases in fluorescence. The close structural analogue of U-73122, U-73343, caused a minimal effect on the concentration-response curve of ET-1. ET-3 produced no major increase of Fluo-3 fluorescence. Removal of extracellular Ca(2+) resulted in a shift to the right of the ET-1 concentration-response curve. Both the L-type voltage-operated Ca(2+) channel blocker, nifedipine, and the ryanodine receptor inhibitor, dantrolene, reduced the efficacy of ET-1. Two protein kinase C inhibitors reduced both potency and efficacy of ET-1. Our results demonstrate that ET(A) receptors are expressed and functionally coupled to rise of [Ca(2+)](i) in H9c2 cardiomyoblasts. ET-1-induced [Ca(2+)](i) increase is triggered by Ca(2+) release from intracellular inositol 1,4,5-trisphosphate-gated stores; plasma membrane Ca(2+) channels and ryanodine receptors participate in sustaining the Ca(2+) response. Regulation of channel opening by protein kinase C is also involved in the process of [Ca(2+)](i) increase.

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Biological Process

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Term	Qualifier	Evidence	With	Reference	Notes	Source	Original Reference(s)
phospholipase C-activating G protein-coupled receptor signaling pathway		IMP		728499		RGD
positive regulation of calcium ion transport		IMP		728499		RGD
positive regulation of release of sequestered calcium ion into cytosol		IMP		728499		RGD
protein kinase C-activating G protein-coupled receptor signaling pathway		IMP		728499		RGD

Objects Annotated

Genes (Rattus norvegicus)

Ednra (endothelin receptor type A)

Objects referenced in this article

Gene	Ednrb	endothelin receptor type B	Rattus norvegicus

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