Expression of multiple plasma membrane Ca(2+)-ATPases in rat pancreatic islet cells.
When stimulated by glucose, the pancreatic beta-cell displays large oscillations of intracellular free Ca2+ concentration ([Ca2+]i). To control [Ca2+]i, the beta-cell must be equipped with potent mechanisms for Ca2+ extrusion. We studied the expression of the plasma membrane Ca(2+)-ATPases (PMCA) in three insulin secreting preparations (a pure beta-cell preparation, RINm5F cells and pancreatic islet cells), using reverse-transcribed PCR, RNase protection assay and Western blotting. The four main isoforms, PMCA1, PMCA2, PMCA3 and PMCA4 were expressed in the three preparations. Six alternative splice mRNA variants, characterized at splice sites A, B and C were detected in the three preparations (rPMCA1xb, 2yb, 2wb, 3za, 3zc, 4xb), plus two additional variants in pancreatic islet cells (PMCA4za, 1xkb). The latter variant corresponded to a novel variant of rat PMCA1 gene lacking the exon coding for the 10th transmembrane segment, at splice site B. At the mRNA and protein level, five variants predominated (1xb, 2wb, 3za, 3zc, 4xb), whilst one additional isoform (4za), predominated at the protein level only. This provides the first evidence for the presence of PMCA2 and PMCA3 isoforms at the protein level in non-neuronal tissue. Hence, the pancreatic beta-cell is equipped with multiple PMCA isoforms with possible differential regulation, providing a full range of PMCAs for [Ca2+]i regulation.
Kamagate
A
A
Laboratory of Pharmacology, Brussels Free University School of Medicine, Belgium.
Herchuelz
A
A
Bollen
A
A
Van Eylen
F
F
D000199:Actins / Q000235:genetics; D000818:Animals; D015153:Blotting, Western; D000252:Calcium-Transporting ATPases / Q000235:genetics* / Q000276:immunology / Q000378:metabolism*; D002462:Cell Membrane / Q000201:enzymology*; D005260:Female; D015971:Gene Expression Regulation, Enzymologic; D007515:Islets of Langerhans / Q000201:enzymology*; D007527:Isoenzymes / Q000235:genetics / Q000378:metabolism; D012326:RNA Splicing; D012333:RNA, Messenger / Q000032:analysis / Q000235:genetics; D051381:Rats; D017208:Rats, Wistar; D020133:Reverse Transcriptase Polymerase Chain Reaction
D000199:Actins; D007527:Isoenzymes; D012333:RNA, Messenger; D000252:Calcium-Transporting ATPases