THE EFFECT OF GLUCOSE AND CALCIUM ON CA2-ADENOSINE TRIPHOSPHATASE IN PANCREATIC-ISLETS ISOLATED FROM A NORMAL AND A NON-INSULIN-DEPENDENT DIABETES-MELLITUS RAT MODEL()

Regulation of calcium balance is important in the secretory function o f pancreatic islets, Ca2+-adenosine triphosphatase (ATPase) is altered in tissues of non-insulin-dependent diabetes mellitus (NIDDM) rats, a nd they have an impaired response to glucose, ''glucose blindness.'' W e propose that the glucose blindness of the diabetic islet is the resu lt of defective cellular calcium metabolism. Since Ca2+-ATPase activit y is important in the regulation of calcium balance, we investigated t he effect of glucose and/or calcium on Ca2+-ATPase activity in pancrea tic islets in vitro and compared it with the effect in freshly isolate d islets from controls and from rats with NIDDM induced by streptozoto cin neonatally. Islets were isolated using collagenase and were stored fresh or cultured up to 2 days in RPMI 1640 in the presence of differ ent concentrations of glucose and calcium, Membrane Ca2+-ATPase activi ty, insulin secretion, and insulin content were determined. Ca2+-ATPas e activity was 1.30 +/- 0.20 mu mol/L Pi/mu g membrane protein in norm al noncultured islets and 1.02 +/- 0.15 in islets cultured in 5.6 mmol /L glucose. Ca2+-ATPase activity progressively decreased to 0.56 +/- 0 .10 and 0.34 +/- 0.15 mu mol/L Pi/mu g membrane protein when glucose w as increased in the culture media to 16.6 and 27.7 mmol/L, respectivel y. Decreasing glucose to 2.8 mmol/L did not alter Ca2+-ATPase activity . Increasing or decreasing the Ca2+ content of the media did not signi ficantly change Ca2+-ATPase activity. Islets isolated from NIDDM rats had lower basal Ca2+-ATPase activity and insulin content compared with normal controls. Incubation of islets from diabetic rats in high gluc ose further decreased the Ca2+-ATPase content, but incubation in low g lucose did not reverse it, Insulin secretion was responsive to glucose and calcium in normal islets, but was suppressed in islets from diabe tic animals, From these studies, we conclude that high glucose, but no t calcium, decreases Ca2+-ATPase activity in islets from normal rats. Islets from NIDDM rats with glucose blindness have decreased Ca2+-ATPa se activity, likely due to the glucose status. We suggest that this de creased Ca2+-ATPase activity may contribute to the pancreatic islets' glucose blindness. Copyright (C) 1998 by W.B. Saunders Company.