Regulation of N- and L-type Ca2+ channels in adult frog sympathetic ganglion B cells by nerve growth factor in vitro and in vivo.

Abstract

To examine mechanisms responsible for the long-term regulation of Ca2+-channels in an adult neuron, changes in whole cell Ba2+ current (IBa) were examined in adult bullfrog sympathetic ganglion B cells in vitro. Cells were cultured at low density in defined, serum free medium. After 15 days, total IBa was similar to the initial value, whereas IBa density was reduced by approximately 36%, presumably due to an increase in neuronal surface area. By contrast, IBa density remained constant after 6-15 days in the presence of murine beta-NGF (200 ng/ml), and total IBa was almost doubled. Inclusion of cytosine arabinoside (Ara-C; 10 microM) to inhibit proliferation of nonneuronal cells, did not affect the survival of neurons in the absence of nerve growth factor (NGF) nor did it attenuate IBa. Ara-C did not prevent the effect of NGF on IBa. There were three independent components to the action of NGF; during 6-9 days, it increased omega-conotoxin-GVIA-sensitive N-type IBa (IBa,N); increased nifedipine-sensitive L-type IBa (IBa,L) and decreased inactivation of the total Ba2+ conductance (gBa). The latter effect involved a selective decrease in the amplitude of one of the four kinetic components that describe the inactivation process. Total IBa was also 55.8% larger than control in the somata of B cells acutely dissociated from leopard frogs that had received prior subcutaneous injections of NGF. By contrast, injection of NGF antiserum decreased total IBa by 29.4%. There was less inactivation of gBa in B cells from NGF-injected animals than in cells from animals injected with NGF antiserum (P < 0.001). These data suggest that NGF-like molecule(s) play(s) a role in the maintenance of IBa in an adult amphibian sympathetic neuron; the presence of NGF may allow the neuron to maintain a constant relationship between cell size and current density. They also show that IBa inactivation in an adult neuron can be modulated in a physiologically relevant way by an extracellular ligand.