Roles of hippocampal nitric oxide and calcium/calmodulin-dependent protein kinase II in inhibitory avoidance learning in rats

Abstract

This study investigated the interactive roles of nitric oxide and calcium/calmodulin-dependent protein kinase II in inhibitory avoidance learning. In Experiment I, rats were trained on a one-trial step-through inhibitory avoidance learning task, whereas the controls were trained in a noncontingent stimulus-pairing condition. The experimental rats showed significantly higher retention scores than the control rats. Correspondingly, the rats in the experimental group showed significantly higher Ca2+-independent activity of the hippocampal calcium/calmodulin-dependent protein kinase II and a significant increase in the endogenous phosphorylation of neuronal nitric oxide synthase. The intrahippocampal infusion of 7-nitro-indazole, 2-[N-(2-hidroxyethyl)-N-(4-methoxy-benzenesulfonyl)]-amino-N-(4-chlorocinnamyl)-N-methylbenzylamine, or 2-amino-5-phosphonopentanoic acid disrupted inhibitory avoidance learning. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis showed that these drugs significantly depressed phosphorylation of hippocampal nitric oxide synthase. The Ca2+-independent activity of hippocampal calcium/calmodulin-dependent protein kinase II was significantly lower in the 2-[N-(2-hidroxyethyl)-N-(4-methoxy-benzenesulfonyl)]-amino-N-(4-chlorocinnamyl)-N-methylbenzylamine or the 2-amino-5-phosphonopentanoic acid-infused group compared with the controls. Although these depressed activities were not reversed by the infusion of a nitric oxide donor (sodium nitroprusside), this did significantly improve the rats' inhibitory avoidance deficit. These results, taken together, indicate that the nitric oxide synthase activation is essential for inhibitory avoidance learning, which may be triggered via the calcium/calmodulin-dependent protein kinase II activation in the hippocampus.