Fibroblast growth factor-induced increases in calcium currents in the PC12 pheochromocytoma cell line are tyrosine phosphorylation dependent.
The PC12 rat pheochromocytoma cell line is widely used to study neuronal differentiation by growth factors. In response to nerve growth factor (NGF) and basic fibroblast growth factor (bFGF), PC12 cells differentiate into sympathetic-like neurons and become electrically excitable. Using whole cell patch-clamp recording, with barium as a charge carrier, we looked at the effects of bFGF on calcium channel expression as reflected by changes in barium current amplitudes normalized to cell membrane area. Similar to the effect reported for NGF, we show that 7 day treatment with bFGF increased the barium current approximately 4-fold. The largest contributor to the increase in barium current with bFGF treatment is a 6-fold increase in the high threshold voltage activated omega-conotoxin sensitive barium current. Smaller increases in current produced by bFGF treatment of PC12 cells are observed for the dihydropyridine sensitive and dihydropyridine/conotoxin insensitive currents. The bFGF-induced increases in barium currents are dependent on tyrosine phosphorylation, since the effects of bFGF are blocked by genistein, a tyrosine kinase inhibitor. This system will ultimately be useful in understanding the signaling pathways that control calcium channel expression in response to growth factors.
Rane
S G
SG
Department of Biological Sciences, Purdue University, West Lafayette, Indiana.
Pollock
J D
JD
D000818:Animals; D001464:Barium / Q000378:metabolism; D001692:Biological Transport / Q000187:drug effects; D002118:Calcium / Q000378:metabolism*; D015220:Calcium Channels / Q000096:biosynthesis* / Q000187:drug effects / Q000235:genetics; D004095:Dihydropyridines / Q000494:pharmacology; D016222:Fibroblast Growth Factor 2 / Q000494:pharmacology*; D019833:Genistein; D007529:Isoflavones / Q000494:pharmacology; D009363:Neoplasm Proteins / Q000235:genetics / Q000378:metabolism; D016716:PC12 Cells / Q000187:drug effects*; D018408:Patch-Clamp Techniques; D010455:Peptides / Q000494:pharmacology; D010766:Phosphorylation; D011499:Protein Processing, Post-Translational; D011505:Protein-Tyrosine Kinases / Q000037:antagonists & inhibitors / Q000502:physiology*; D051381:Rats; D015398:Signal Transduction / Q000502:physiology*; D015854:Up-Regulation / Q000502:physiology*; D020866:omega-Conotoxin GVIA
D015220:Calcium Channels; D004095:Dihydropyridines; D007529:Isoflavones; D009363:Neoplasm Proteins; D010455:Peptides; D016222:Fibroblast Growth Factor 2; D001464:Barium; C038806:1,4-dihydropyridine; D020866:omega-Conotoxin GVIA; D019833:Genistein; D011505:Protein-Tyrosine Kinases; D002118:Calcium