Modulation of endothelin-A receptor, Galpha subunit, and RGS2 expression during H9c2 cardiomyoblast differentiation.
In cardiac myocytes, growth responses depend on activation of G protein-coupled receptors interacting with Gq/11 protein subfamily members. Endothelin receptors of the ETA subtype belong to this receptor group inducing hypertrophic responses. To understand the role of ETA receptors and signal transduction proteins in modulating cell growth, we analyzed the pharmacological profile of this receptor, its level of expression together with those of Galpha subunits and the RGS2 protein in cardiomyoblasts differentiating into the cardiac phenotype. H9c2 rat cardiomyoblasts were grown in the presence of 10% fetal bovine serum (FBS) or 1% FBS plus all-trans-retinoic acid to induce the cardiac phenotype. The pharmacological properties of ETA receptors were investigated by competition-binding experiments, whereas the protein expression profile was analyzed by immunoblot and immunocytochemistry. The pharmacological profile of ETA receptors changed during differentiation of cardiomyoblasts into cardiomyocytes, and the amount of expressed receptor appeared to increase. Immunocytochemistry also showed a marked increase of receptor expression on cell membranes of differentiated cardiomyocytes. Among the other signaling proteins examined, both Galphaq/11 and RGS2 expression decreased in cells with the cardiac phenotype. Our results demonstrate that the expression of key proteins (ETA receptor, Galphaq/11, and RGS2) involved in signal transduction of hypertrophic stimuli is modulated during cell differentiation and correlates with the cardiac phenotype.
Giusti
Laura
L
Dipartimento di Psichiatria, Neurobiologia, Farmacologia e Biotecnologie, Università di Pisa, Pisa, Italy.
Gargini
Claudia
C
Ceccarelli
Francesca
F
Bacci
Michela
M
Italiani
Paola
P
Mazzoni
Maria R
MR
D000818:Animals; D001667:Binding, Competitive; D015153:Blotting, Western; D002454:Cell Differentiation; D002462:Cell Membrane / Q000378:metabolism; D049109:Cell Proliferation; D002478:Cells, Cultured; D003470:Culture Media / Q000494:pharmacology; D019084:Fluorescent Antibody Technique, Indirect; D043802:GTP-Binding Protein alpha Subunits, Gq-G11 / Q000096:biosynthesis*; D006801:Humans; D015151:Immunoblotting; D007150:Immunohistochemistry; D007700:Kinetics; D008024:Ligands; D032383:Myocytes, Cardiac / Q000166:cytology* / Q000378:metabolism; D010641:Phenotype; D011485:Protein Binding; D020710:RGS Proteins / Q000096:biosynthesis*; D051381:Rats; D044022:Receptor, Endothelin A / Q000096:biosynthesis*; D015398:Signal Transduction
D003470:Culture Media; D008024:Ligands; D020710:RGS Proteins; D044022:Receptor, Endothelin A; C105345:Rgs2 protein, mouse; D043802:GTP-Binding Protein alpha Subunits, Gq-G11