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Diabetes 51:366-375, 2002
© 2002 by the American Diabetes Association, Inc.

Overexpression of the Na/Ca Exchanger Shapes Stimulus-Induced Cytosolic Ca2+ Oscillations in Insulin-Producing BRIN-BD11 Cells

Françoise Van Eylen1, Oscar Diaz Horta1, Aurore Barez1, Adama Kamagate1, Peter R. Flatt2, Regina Macianskiene3, Kanigula Mubagwa3, and André Herchuelz1

1 Laboratory of Pharmacology, Brussels University School of Medicine, Brussels, Belgium
2 School of Biomedical Sciences, University of Ulster, Coleraine, Northern Ireland, U.K.
3 Centre of Experimental Surgery and Anesthesiology, University of Leuven, Leuven, Belgium

In response to glucose, mouse ß-cells display slow oscillations of the membrane potential and cytosolic free Ca2+ concentration ([Ca2+]i), whereas rat ß-cells display a staircase increase in these parameters. Mouse and rat islet cells differ also by their level of Na/Ca exchanger (NCX) activity. The view that the inward current generated by Na/Ca exchange shapes stimulus-induced electrical activity and [Ca2+]i oscillations in pancreatic ß-cells was examined in insulin-producing BRIN-BD11 cells overexpressing the Na/Ca exchanger. BRIN-BD11 cells were stably transfected with NCX1.7, one of the exchanger isoforms identified in the ß-cell. Overexpression could be assessed at the mRNA and protein level. Appropriate targeting to the plasma membrane could be assessed by microfluorescence and the increase in Na/Ca exchange activity. In response to K+, overexpressing cells showed a more rapid increase in [Ca2+]i on membrane depolarization as well as a more rapid decrease of [Ca2+]i on membrane repolarization. In response to glucose and tolbutamide, control BRIN cells showed large amplitude [Ca2+]i oscillations. In contrast, overexpressing cells showed a staircase increase in [Ca2+]i without such large oscillations. Diazoxide-induced membrane hyperpolarization restored large amplitude [Ca2+]i oscillations in overexpressing cells. The present data confirm that Na/Ca exchange plays a significant role in the rat ß-cell [Ca2+]i homeostasis, the exchanger being a versatile system allowing both Ca2+ entry and outflow. Our data suggest that the current generated by the exchanger shapes stimulus-induced membrane potential and [Ca2+]i oscillations in insulin-secreting cells, with the difference in electrical activity and [Ca2+]i behavior seen in mouse and rat ß-cells resulting in part from a difference in Na/Ca exchange activity between these two cells.


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