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Research ArticleArticle

Specific Gα11β3γ5Protein Involvement in Endothelin Receptor-Induced Phosphatidylinositol Hydrolysis and Ca2+ Release in Rat Portal Vein Myocytes

Nathalie Macrez, Jean-Luc Morel and Jean Mironneau
Molecular Pharmacology April 1999, 55 (4) 684-692;
Nathalie Macrez
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Jean-Luc Morel
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Jean Mironneau
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Abstract

In this study, we identified the receptor subtype activated by endothelin-1 (ET-1) and the subunit composition of the G protein coupling this receptor to increase in cytosolic Ca2+concentration in rat portal vein myocytes. We used intranuclear antisense oligonucleotide injection to selectively inhibit the expression of G protein subunits. We show here that the endothelin receptor subtype A (ETA)-mediated increase in cytosolic Ca2+ concentration was mainly dependent on Ca2+ release from the intracellular store. ETAreceptor-mediated Ca2+ release was selectively inhibited by antisense oligonucleotides that inhibited the expression of α11, β3, and γ5 subunits, as checked by immunocytochemistry. Intracellular dialysis of a carboxyl terminal anti-βcom antibody and a peptide corresponding to the Gβγ binding region of the β-adrenergic receptor kinase-1 had no effect on the ETAreceptor-mediated Ca2+ release. In contrast, a synthetic peptide corresponding to the carboxyl terminus of the αq/α11 subunit, heparin (an inhibitor of inositol 1,4,5-trisphosphate receptors), and U73122 (an inhibitor of phosphatidylinositol-phospholipase C) inhibited, in a concentration-dependent manner, the ETA receptor-mediated Ca2+ responses. Accumulation of [3H]inositol trisphosphate evoked by norepinephrine peaked at ∼15 s, whereas that evoked by ET-1 progressively increased within 2 min. In myocytes injected with anti-αq antisense oligonucleotides, both amplitude and time course of the norepinephrine-induced Ca2+ release became similar to those of the ET-1-induced Ca2+ response. We conclude that the ETAreceptor-mediated Ca2+ release is selectively transduced by the heterotrimeric G11 protein composed of α11, β3, and γ5 subunits, and that a delayed stimulation of phospholipase C occurs via the α11 subunit.

Footnotes

  • Send reprint requests to: Dr J. Mironneau, Laboratoire de Physiologie Cellulaire et Pharmacologie Moléculaire, CNRS ESA 5017, Université de Bordeaux II, 146 rue Léo Saignat, 33076 Bordeaux Cedex, France. Email: jean.mironneau{at}esa5017.u-bordeaux2.fr

  • ↵1 Contributed equally to the work.

  • This work was supported by a grant from the Centre National de la Recherche Scientifique (France).

  • Abbreviations:
    ET-1
    endothelin-1
    AII
    angiotensin II
    βARK1
    β-adrenergic receptor kinase-1
    [Ca2+]i
    cytosolic Ca2+concentration
    InsP
    inositol phosphate
    InsP1
    inositol monophosphate
    InsP2
    inositol bisphosphate
    InsP3
    inositol trisphosphate
    NE
    norepinephrine
    PLC
    phospholipase C
    ETA
    endothelin receptor subtype A
    ETB
    endothelin receptor subtype B
    U73122
    1-(6-((17β-3 methoxystra 1,3,5 (10)-trien-17-yl) amino) hexyl)-1H-pyrrole-2,5-dione
    PTX
    pertussis toxin
    AM
    indo-1-acetoxymethylester
    • Received June 22, 1998.
    • Accepted January 9, 1999.
  • The American Society for Pharmacology and Experimental Therapeutics
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Molecular Pharmacology: 55 (4)
Molecular Pharmacology
Vol. 55, Issue 4
1 Apr 1999
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Research ArticleArticle

Specific Gα11β3γ5Protein Involvement in Endothelin Receptor-Induced Phosphatidylinositol Hydrolysis and Ca2+ Release in Rat Portal Vein Myocytes

Nathalie Macrez, Jean-Luc Morel and Jean Mironneau
Molecular Pharmacology April 1, 1999, 55 (4) 684-692;

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Research ArticleArticle

Specific Gα11β3γ5Protein Involvement in Endothelin Receptor-Induced Phosphatidylinositol Hydrolysis and Ca2+ Release in Rat Portal Vein Myocytes

Nathalie Macrez, Jean-Luc Morel and Jean Mironneau
Molecular Pharmacology April 1, 1999, 55 (4) 684-692;
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