Effect of ethanol on intracellular ionized calcium concentrations in synaptosomes and hepatocytes.
The effect of ethanol on intracellular ionized calcium concentrations (Cai) was studied in synaptosomes isolated from mouse whole brain and in hepatocytes isolated from rat liver. The fluorescent calcium chelator, fura-2, was used to quantitate Cai. Incubation of synaptosomes with ethanol (350-700 mM) increased resting Cai and decreased the effectiveness of KCl to raise Cai in a concentration-dependent manner. Ethanol produced an initial rapid (less than 10 sec) increase in resting Cai that remained elevated for at least 14 min in the presence of the drug. The increase in resting Cai was correlated with the inhibitory effect of ethanol on depolarization-induced increases in Cai. Resting Cai was dependent on the external calcium concentration (0-1 mM). However, the ethanol-induced increase in resting Cai (expressed as percent of control) did not differ in the presence of several extracellular calcium concentrations (0.01, 0.1, and 1 mM). Incubation of synaptosomes in a Na-free buffer resulted in a higher resting Cai and slightly enhanced the effect of ethanol to increase resting Cai. In contrast to these results in brain tissue, ethanol (30-600 mM) did not alter resting Cai or vasopressin-stimulated increases in Cai in hepatocytes. Our results suggest that the anesthetic effects of alcohols may be mediated, in part, by increased resting Cai and by decreased calcium influx through voltage-sensitive calcium channels. In addition, our findings suggest possible mechanisms by which ethanol increases resting Cai in neuronal tissue.
Daniell
L C
LC
Department of Pharmacology, University of Colorado Health Sciences Center, Denver 80262.
Brass
E P
EP
Harris
R A
RA
D000818:Animals; D002118:Calcium / Q000032:analysis* / Q000378:metabolism; D000431:Ethanol / Q000494:pharmacology*; D008099:Liver / Q000032:analysis / Q000187:drug effects*; D008297:Male; D051379:Mice; D008813:Mice, Inbred ICR; D011188:Potassium / Q000494:pharmacology; D012964:Sodium / Q000378:metabolism; D013574:Synaptosomes / Q000032:analysis / Q000187:drug effects*
D000431:Ethanol; D012964:Sodium; D011188:Potassium; D002118:Calcium