Purinergic stimulation of rat cardiomyocytes induces tyrosine phosphorylation and membrane association of phospholipase C gamma: a major mechanism for InsP3 generation.
Phospholipase C gamma (PLC gamma) expression and activation by a purinergic agonist were investigated in adult rat cardiomyocytes. PLC gamma is expressed in isolated cardiomyocytes. Stimulation of cells with extracellular ATP induces a rapid increase in membrane-associated PLC gamma immunoreactivity most probably due to redistribution of the lipase from the cytosol to the membrane. The purine triggers a significant phosphorylation on tyrosine residues of a cytosolic pool of PLC gamma with a time course that correlates with that of translocation. Extracellular ATP also increases intracellular Ins(1,4,5)P3 content. All these events (translocation and phosphorylation of PLC gamma, InsP3 formation) are blocked by genistein, a tyrosine kinase inhibitor. The purinergic effect on both PLC gamma translocation and phosphorylation are Ca-sensitive. We thus propose that the purinergic stimulation activates a non-receptor tyrosine kinase that phosphorylates PLC gamma in the presence of an increased Ca level and induces PLC gamma redistribution to the membrane. There, PLC gamma becomes activated leading to the hydrolysis of phosphatidylinositol diphosphate and in turn Ins(1,4,5)P3 formation. This cascade of events may play a significant role in the induction of arrhythmogenesis by purinergic agonists.
Puceat
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INSERM U-390, Laboratoire de Physiopathologie Cardiovasculaire, C.H.U. Amaud de Villeneuve, Montpellier, France.
Vassort
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D000255:Adenosine Triphosphate / Q000378:metabolism / Q000494:pharmacology*; D000818:Animals; D015153:Blotting, Western; D002462:Cell Membrane / Q000201:enzymology; D004789:Enzyme Activation; D004791:Enzyme Inhibitors / Q000494:pharmacology; D015870:Gene Expression; D019833:Genistein; D066298:In Vitro Techniques; D015544:Inositol 1,4,5-Trisphosphate / Q000378:metabolism*; D007527:Isoenzymes / Q000302:isolation & purification / Q000378:metabolism*; D007529:Isoflavones / Q000494:pharmacology; D007700:Kinetics; D008297:Male; D009206:Myocardium / Q000378:metabolism*; D051966:Phospholipase C gamma; D010750:Phosphoproteins / Q000302:isolation & purification / Q000378:metabolism*; D010766:Phosphorylation; D019000:Phosphotyrosine / Q000378:metabolism*; D011505:Protein-Tyrosine Kinases / Q000037:antagonists & inhibitors / Q000378:metabolism*; D051381:Rats; D017208:Rats, Wistar; D010738:Type C Phospholipases / Q000302:isolation & purification / Q000378:metabolism*
D004791:Enzyme Inhibitors; D007527:Isoenzymes; D007529:Isoflavones; D010750:Phosphoproteins; D019000:Phosphotyrosine; D015544:Inositol 1,4,5-Trisphosphate; D000255:Adenosine Triphosphate; D019833:Genistein; D011505:Protein-Tyrosine Kinases; D010738:Type C Phospholipases; D051966:Phospholipase C gamma