P2‐purinoceptor‐stimulated phosphoinositide turnover in chick myotubes calcium mobilization and the role of guanyl nucleotide‐binding proteins

Abstract

ATP, a trigger of P2-purinoceptor-mediated polyphosphoinositide (PI) turnover in cultured myotubes, increased cytosolic calcium levels in a time- and dose-dependent manner (quin2 fluorescence). The calcium was released from intracellular stores, as acute addition of 5 mM EGTA was without significant effect. Adenosine 5′-(3-thiotriphosphate) and 5′-adenylyl imidodiphosphate also increased intracellular levels of inositol phosphates (InsP) and cytosolic calcium levels. Treatment with cholera or pertussis toxin of myotube cultures did not affect the P2-purinoceptor-mediated InsP increase although PI turnover in permeabilized myotubes was stimulated by guanosine 5′-(3-thiotriphosphate). The results suggest that myotube P2-purinoceptors trigger PI turnover and increase intracellular free calcium levels, via a mechanism insensitive to ADP-ribosylation, by cholera or pertussis toxin of guanyl nucleotide-binding (G) proteins. However, the presence of a phospholipase C-coupled G-protein was otherwise demonstrated.