CANNABINOIDS INHIBIT N-TYPE AND P Q-TYPE CALCIUM CHANNELS IN CULTUREDRAT HIPPOCAMPAL-NEURONS/

Cannabinoids and their analogues have been found to inhibit N- and P/Q -type Ca2+ currents in cell lines and sympathetic neurons transfected with cannabinoid CB1 receptor. However, the effects of cannabinoids on Ca2+ currents in the CNS are largely unexplored. In this study we inv estigated whether these compounds inhibit Ca2+ channels in cultured ra t hippocampal neurons. With the use of antibodies directed against the amino-terminus of the CB1 receptor, we found that in 5-day cultures p yramidally shaped neurons expressed somatic CB1 receptors, whereas in 4-wk cultures the receptor was predomintely located on neurites. In ea rly cultures, the cannabimimetic WIN 55,212-2 reversibly inhibited who le cell Ba2+ current in a concentration-dependent (K-1/2 = 21 nM) and pertussis-toxin-sensitive fashion. Inhibition was reduced by the CB1 a ntagonist SR141716. The current was unaffected by the nonpsychoactive enantiomer WIN 55,212-3. Maximal inhibition by the nonclassical cannab inoid agonist CP 55,940 and by an endogenous cannabinoid, anandamide, were similar to that seen with maximal concentrations of WIN 55,212-2. The Ba2+ current modulated by cannabinoids was carried by N-type (ome ga-conotoxin-GVIA-sensitive) and P/Q-type (omega-conotoxin-MVIIC-sensi tive) channels. These results demonstrate cannabinoid-receptor-mediate d inhibition of distinct Ca2+ channels in central neurons. Because the channels that underlie these currents are chiefly located presynaptic ally, and are required for evoked neurotransmitter release, our result s suggest a major role for cannabinoids (endogenous and exogenous) in the modulation of synaptic transmission at CNS synapses.