Effects of cationic modification on 45Ca uptake and insulin release by transplantable rat insulinoma cells maintained in tissue culture

Abstract

1. Acute effects of cations on 45Ca uptake and insulin release by transplantable rat insulinoma cells were examined after 2-3 days culture in RPMI-1640 containing 11.1 mM glucose. 2. At 2.6 mM Ca2+, rat insulinoma cells (greater than 95% viability) released 78-158 ng insulin/10(6) cells during 60 min incubation with uptake at 2.19-3.24 nmol 45Ca/10(6) cells. 3. Addition of 2 mM La3+, Co2+, Mn2+, Zn2+ or Ba2+ did not affect 45Ca uptake. Insulin release was also unaffected by these cations with the exception of 87% inhibition in the presence of La3+ or Zn2+. 4. Omission of 5.9 mM K+, 1.2 mM Mg2+, 115 mM Na+ or H+ (pH 8.5) did not affect 45Ca uptake or insulin release, irrespective of osmotic compensation using choline chloride or sucrose. Rat insulinoma cells were similarly unresponsive to addition of 30.9 mM K+, 12 mM Mg2+ or H+ (pH 6.3). 5. Omission of 2.6 mM Ca2+ (with or without addition of 1 mM EGTA) or addition of 20.5 mM Ca2+ did not affect insulin release. 6. The results indicate that rat insulinoma cells are little affected by cationic modifications which have profound effects on Ca2+ handling and insulin release by pancreatic beta-cells. Dysregulation of insulin release by insulinoma cells is associated with marked irregularities in the control of transmembrane Ca2+ fluxes and sensitivity to extracellular Ca2+.