Identification, expression pattern and potential activity of Na/Ca exchanger isoforms in rat pancreatic B-cells.
In the pancreatic B-cell, Na/Ca exchange displays a quite high capacity and participates in the control of cytosolic free Ca2+ concentration. The Na/Ca exchanger was recently cloned in various tissues. Two genes coding for two different exchangers (NCX1 and NCX2) have been identified and evidence for several isoforms for NCX1 shown. To characterize the isoform(s) expressed in pancreatic B-cells, a RT-PCR analysis was performed on mRNA from rat pancreatic islets, purified B-cells and insulinoma B-cells (RINm5F cells). PCR amplification did not yield the expected NCX2 DNA fragment but yielded 2 NCX1 bands, corresponding to NaCa3 and NaCa7, in the three preparations. NaCa3 and NaCa7 were equally expressed in pancreatic islets and purified B-cells. In RINm5F cells, NaCa3 expression did not differ from that in islet and purified B-cells but NaCa7 was 3 times less expressed. This lower expression was accompanied by a 3 times lower Na/Ca exchange activity in RINm5F cells compared to islet cells. Our data indicate the existence of 2 NCX1 isoforms but not of NCX2 in pancreatic B-cells. The difference in both the expression patterns of NCX1 isoforms and the activity of Na/Ca exchange in islet cells and RINm5F cells is compatible with a difference in activity between NaCa3 and NaCa7.
Van Eylen
F
F
Laboratory of Pharmacology, Brussels University School of Medicine, Belgium.
Svoboda
M
M
Herchuelz
A
A
D000818:Animals; D002118:Calcium / Q000378:metabolism; D002352:Carrier Proteins / Q000235:genetics / Q000378:metabolism*; D002478:Cells, Cultured; D007328:Insulin / Q000378:metabolism; D007515:Islets of Langerhans / Q000166:cytology* / Q000378:metabolism*; D007527:Isoenzymes / Q000378:metabolism; D016133:Polymerase Chain Reaction / Q000379:methods; D051381:Rats; D011919:Rats, Inbred Strains; D019831:Sodium-Calcium Exchanger; D014018:Tissue Distribution
D002352:Carrier Proteins; D007328:Insulin; D007527:Isoenzymes; D019831:Sodium-Calcium Exchanger; D002118:Calcium