Cholinergic excitation of dopaminergic cells depends on sequential activation of protein kinase C and the L-type calcium channel in ventral tegmental area slices

Dopaminergic projections from the ventral tegmental area (VTA) constitute the mesolimbocortical system that underlies addiction and psychosis primarily as the result of increased dopaminergic transmission. Dopaminergic neurons in the VTA receive glutamatergic and cholinergic innervations that regulate their firing activities. Both transmitter systems can activate protein kinase C (PKC) by increasing intracellular calcium and lipid second messengers, however, whether PKC mediates increased firing following glutamatergic and cholinergic activation remains unknown. This paper examined the effects of acute PKC inhibition on firing responses to carbachol, NMDA or AMPA using patch clamp recordings from brain slices. The three ligands all induced a reversible increase in firing, however, only carbachol-induced increase in firing was attenuated by the PKC inhibitors chelerythrine or GF 109203X. The L-type calcium channel blocker nifedipine partially blocked carbachol-induced excitation similar to PKC inhibitors. PKC inhibition and L-type channel blockade did not significantly alter NMDA- or AMPA-induced excitation. Concurrent blockade of PKC and L-type channels with chelerythrine and nifedipine did not additively suppress carbachol-induced excitation indicating they were sequential events in the same signaling pathway. Furthermore, preincubation with the PKC inhibitor GF 109203X reduced the carbachol-induced increase in nifedipine-sensitive high-voltage gated calcium currents. These results indicate that cholinergic activation enhances PKC activity, which in turn facilitates L-type channel opening to excite dopaminergic cells, a finding that is in line with reports of increased PKC in the VTA in animals displaying addictive behavior.