The effect of antisense oligonucleotide treatment of plasma membrane Ca(+2)-ATPase in PC12 cells.
Plasma membrane Ca(+2)-ATPase (PMCA), encoded by four separate genes, constitutes a high affinity system extruding Ca(+2) outside the cell. The nerve growth factor-treated PC12 cell line possesses all four main PMCA isoforms. To evaluate the potential role of PMCA isoforms in the differentiation process, we transiently suppressed the expression of PMCA2 and 3 using the antisense oligonucleotides. In the transfected PC12 cells, we observed morphological changes, slowed neurite extension and diminished survival of the cells. The apparent transport activity and affinity of the calcium pump to Ca(+2) were lower in the cells with suppressed PMCA2 and 3 isoforms than in the control cells. Moreover, in the transfected PC12 plasma membranes, the calcium pump was insensitive to stimulation by calmodulin. These findings suggest that PMCA2 and 3 isoforms may be involved in developmental and differentiation processes.
Neurochemical Laboratory, Department of Biochemistry, Medical University, ul. Mazowiecka 6/8, 92-215 Łódź, Poland.
Janusz
J
Szemraj
Iwona
I
Kawecka
Jacek
J
Bartkowiak
Ludmiła
L
Zylińska
D000818:Animals; D002118:Calcium / Q000378:metabolism*; D000252:Calcium-Transporting ATPases / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*; D002147:Calmodulin / Q000494:pharmacology; D002454:Cell Differentiation / Q000187:drug effects / Q000502:physiology; D002462:Cell Membrane / Q000235:genetics / Q000378:metabolism*; D002470:Cell Survival / Q000187:drug effects / Q000502:physiology; D007527:Isoenzymes / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*; D009414:Nerve Growth Factors / Q000378:metabolism; D016501:Neurites / Q000378:metabolism; D016376:Oligonucleotides, Antisense / Q000235:genetics / Q000378:metabolism*; D016716:PC12 Cells; D051381:Rats
D002147:Calmodulin; D007527:Isoenzymes; D009414:Nerve Growth Factors; D016376:Oligonucleotides, Antisense; D000252:Calcium-Transporting ATPases; D002118:Calcium