Nerve growth factor-mediated stimulation of tyrosine hydroxylase activity in a clonal rat pheochromocytoma cell line

Abstract

In order to confirm the multiple neurotransmitter biosynthetic ability, the possibility of separation of the activities of tyrosine hydroxylase (TH), choline acetyltransferase and glutamic acid decar☐ylase was tested by subcloning of a clonal rat pheochromocytoma PC12 cell line. All of 9 subclones obtained showed significant activities of above 3 enzymes, indicating that the PC12 cell has multi-functional properties of neurotransmitter syntheses. One of the subclones, designated PC12h, was demonstrated to have nerve growth factor- (NGF) responsive TH activity. The ED 50 value of NGF to increase the TH activity was 1.7 ng/ml ( 6.5 × 10 −11M ). A simultaneous addition of saturating amounts of NGF (50 ng/ml) and dexamethasone (10 −6 M) resulted in the increase of TH activity that is equal to the sum of those achieved when either effector was added separately, indicating that the NGF-mediated increase of TH activity in PC12h cells was independent upon the effect of dexamethasone. And also, the TH activity increased by NGF was somewhat potentiated in PC12h cells cultured in a hormone-supplemented serum-free medium.