CA CALMODULIN-DEPENDENT KINASE-II INHIBITOR KN62 ATTENUATES GLUTAMATERELEASE BY INHIBITING VOLTAGE-DEPENDENT CA2+-CHANNELS/

The effect of KN62 nolinesulphonyl)-N-methyl-L-tyrosyl]-4-phenylpiper azine), a putative inhibitor of Ca/calmodulin-dependent kinase II (Ca/ CaM-K II), on glutamate release from isolated nerve-terminals (synapto somes) was examined. The drug caused a potent inhibition of KCl- and 4 -aminopyridine-evoked glutamate release from isolated nerve-terminals (synaptosomes). Examination of the effect of the inhibitor on Ca2+-inf lux revealed that the diminution of glutamate release could be attribu ted to a decrease in cytosolic Ca. A direct effect of KN62 on synaptos omal Ca2+-channels was confirmed in experiments where Ba, which does n ot support CaM-dependent processes, was used in place of Ca. Additiona lly, whole-cell patch-clamping of cerebellar granule neurones directly demonstrated inhibition of Ca-currents by KN62. We therefore suggest that, in cellular systems, conclusions based on the use of KN62 as a C a/CaM-K II blocker may be ambiguous and should be viewed with caution unless the effect of the drug on Ca-influx has also been quantified. T he effect of KN62 on Ca2+-influx appears to be specific to slowly- or non-inactivating conductances, and therefore presents KN62 as a potent ially useful tool in this context.