Effects of cellular Ca2+ depletion on phospholipid turnover and glycogen phosphorylase a in rat hepatocytes.
The influences of changes in cellular Ca2+ level on membrane phospholipid turnover and cellular function (monitored by glycogen phosphorylase a activity) were investigated in vasopressin- and ionophore A23187-stimulated rat hepatocytes. Addition of vasopressin or A23187 to rat hepatocytes in the presence of extracellular Ca2+ enhanced the phosphorylase a activity by 3 to 4-fold within 1 min, returning to initial activity with further incubation. There was the marked generation of 1,2-diacylglycerol resulted from phospholipase C activation, which followed the transient activation of phosphorylase a. When the incorporation of [32P]phosphate into phospholipids was examined, phosphatidylinositol (PI) labeling due to vasopressin-stimulation remained rather unchanged up to 5 min but then rose gradually. On the other hand, A23187 had little effect on the incorporation into phosphatidylinositol although marked phosphatidic acid (PA) labeling was consequently produced, showing inhibitory effect on the conversion of PA to PI. Deprivation of extracellular Ca2+, which also reduced slightly the intracellular Ca2+ from 3.33 micrograms to 1.38 micrograms/10(7) cells, suppressed but not abolished stimuli-induced phosphorylase a activation without affecting the enhancement of phospholipid metabolism. Hepatocytes depleted of intracellular Ca2+ (0.50 microgram/10(7) cells) no longer showed both phosphorylase a activation and the enhancement of phospholipid metabolism. These findings seem to indicate that phosphorylase a activity is more sensitive than membrane phospholipid turnover to changes of intracellular Ca2+ concentration. The results demonstrate that marked and selective changes in membrane phospholipids depending on the type of stimulants occur upon stimulation of hepatocytes and provide the possibility that these reactions do not trigger glycogen phosphorylase a activation through Ca2+ mobilization.
D000818:Animals; D016718:Arachidonic Acid; D001095:Arachidonic Acids / Q000378:metabolism; D000001:Calcimycin / Q000494:pharmacology; D002118:Calcium / Q000378:metabolism*; D004075:Diglycerides / Q000378:metabolism; D008099:Liver / Q000187:drug effects / Q000378:metabolism*; D008297:Male; D010710:Phosphates / Q000378:metabolism; D010743:Phospholipids / Q000378:metabolism*; D010762:Phosphorylase a / Q000378:metabolism*; D006005:Phosphorylases / Q000378:metabolism*; D051381:Rats; D014667:Vasopressins / Q000494:pharmacology
C011439:1,2-diacylglycerol; D001095:Arachidonic Acids; D004075:Diglycerides; D010710:Phosphates; D010743:Phospholipids; D014667:Vasopressins; D016718:Arachidonic Acid; D000001:Calcimycin; D010762:Phosphorylase a; D006005:Phosphorylases; D002118:Calcium