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J. Biol. Chem., Vol. 275, Issue 37, 28682-28687, September 15, 2000

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Long Term Depression in the CA1 Field Is Associated with a Transient Decrease in Pre- and Postsynaptic PKC Substrate Phosphorylation^*

Geert M. J. Ramakers, Klaartje Heinen, Willem-Hendrik Gispen, and Pierre N. E. de Graan

From the Rudolf Magnus Institute for Neurosciences, Department of Medical Pharmacology, Universiteitsweg 100, 3584 CG Utrecht, The Netherlands

Induction of homosynaptic long term depression (LTD) in the CA1 field of the hippocampus is thought to require activation of N-methyl-D-aspartate receptors, an elevation of postsynaptic Ca²⁺ levels, and a subsequent increase in phosphatase activity. To investigate the spatial and temporal changes in protein phosphatase activity following LTD induction, we determined the in situ phosphorylation state of a pre- (GAP-43/B-50) and postsynaptic (RC3) protein kinase C substrate during N-methyl-D-aspartate receptor-dependent LTD in the CA1 field of rat hippocampal slices. We show that LTD is associated with a transient (<30 min) and D-AP5-sensitive reduction in GAP-43/B-50 and RC3 phosphorylation and that LTD is prevented by the phosphatase inhibitors okadaic acid and cyclosporin A. Our data provide strong evidence for a transient increase in pre- and postsynaptic phosphatase activity during LTD. Since the in situ phosphorylation of the calmodulin-binding proteins GAP-43/B-50 and RC3 changes during both LTD and long term potentiation, these proteins may form part of the link between the Ca²⁺ signal and Ca²⁺/calmodulin-dependent processes implicated in long term potentiation and LTD.

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