INVOLVEMENT OF PROTEIN KINASE C AND PROTEIN KINASE A IN THE ENHANCEMENT OF L-TYPE CALCIUM CURRENT BY GABAB RECEPTOR ACTIVATION IN NEONATAL HIPPOCAMPUS

In the early neonatal period activation of GABA(B) receptors attenuates calcium current through N-type calcium channels while enhancing current through L-type calcium channels in rat hippocampal neurons. The attenuation of N-type calcium current has been previously demonstrated to occur through direct interactions of the beta gamma subunits of G(i/o) G-proteins, but the signal transduction pathway for the enhancement of L-type calcium channels in mammalian neurons remains unknown. In the present study, calcium currents were elicited in acute cultures from postnatal day 6-8 rat hippocampi in the presence of various modulators of protein kinase A (PKA) and protein kinase C (PKC) pathways. Overnight treatment with an inhibitor of G(i/o) (pertussis toxin, 200 ng/ml) abolished the attenuation of calcium current by the GABA(B) agonist, baclofen (10 mu M) with no effect on the enhancement of calcium current. These data indicate that while the attenuation of N-type calcium current is mediated by the G(i/o) subtype of G-protein, the enhancement of L-type calcium current requires activation of a different G-protein. The enhancement of the sustained component of calcium current by baclofen was blocked by PKC inhibitors, GF-109203X (500 nM), chelerythrine chloride (5 mu M), and PKC fragment 19-36 (2 mu M) and mimicked by the PKC activator phorbol-12-myristate-13-acetate (1 mu M). The enhancement of the sustained component of calcium current was blocked by PKA inhibitors H-89 (1 mu M) and PKA fragment 6-22 (500 nM) but not Rp-cAMPS (30 mu M) and it was not mimicked by the PKA activator, 8-Br-cAMP (500 mu M-1 mM). The data suggest that activation of PKC alone is sufficient to enhance L-type calcium current but that PKA may also be involved in the GABA(B) receptor mediated effect. (C) 2011 IBRO. Published by Elsevier Ltd. All rights reserved.