45Ca2+ uptake by dispersed pancreatic islet cells: effect of D-glucose and the calcium probe, chlorotetracycline

Abstract

Uptake of 45Ca2+ was studied in dispersed pancreatic islet cells from non-inbred ob/ob-mice. Like whole islets the dispersed cells responded to 20 mM D-glucose with a markedly increased 45Ca2+-labeling of both the lanthanum-nondisplaceable and the lanthanum-displaceable calcium pools. The pronounced effect of D-glucose could not be reproduced with 3-O-methyl-D-glucose, L-glucose, D-mannose, L-leucine, or D-leucine; however, 45Ca2+ uptake was greater in the presence of L-leucine as compared with D-leucine. 45Ca2+ uptake by dispersed cells or whole islets was stimulated severalfold by 100 microM or more chlorotetracycline. At the concentration of only 10 microM, chlorotetracycline had no effect on whole islets and partially inhibited 45Ca2+ uptake by the dispersed cells. The ability of D-glucose to stimulate 45Ca2+ uptake by islets or dispersed cells remained in the presence of 10 microM chlorotetracycline. Islet cell suspensions apparently represent a valid model for studying how Ca2+ interacts with the cells. However, when using chlorotetracycline as fluorescent Ca2+ probe, attention must be paid to it potential ionophoric activity. At only 10 microM, the drug seems to monitor a peripheral pool of Ca2+, some of which may reside in normal transport channels.