N-methyl- d-aspartate receptor-dependent long-term potentiation in CA1 region affects synaptic expression of glutamate receptor subunits and associated proteins in the whole hippocampus

Abstract

Long term potentiation in hippocampus, evoked by high-frequency stimulation, is mediated by two major glutamate receptor subtypes, α-amino-3-hydroxyl-5-methyl-4-isoxazole propionate receptors and N-methyl- d-aspartate receptors. Receptor subunit composition and its interaction with cytoplasmic proteins constitute different pathways regulating synaptic plasticity. Here, we provide further evidence that N-methyl- d-aspartate receptor-mediated long term potentiation evoked at hippocampal CA1 region of rats induced by high-frequency stimulation of the Schaffer collateral-commissural pathway in vivo is not dependent on N-methyl- d-aspartate receptor subunit NR2B. Applying semi-quantitative immunoblotting, we found that in the whole tetanized hippocampus, synaptic expression of the N-methyl- d-aspartate and α-amino-3-hydroxyl-5-methyl-4-isoxazole propionate receptor subunits (NR1, NR2A, glutamate receptor 1) and their associated partners, e.g. synaptic associated protein 97, postsynaptic density protein 95, α subunit of Ca 2+/calmodulin-dependent protein kinase II, neuronal nitricoxide synthase, increased 180 min post-high-frequency stimulation. Moreover, phosphorylation of Ca 2+/calmodulin-dependent protein kinase II at thr286 and glutamate receptor 1 at ser831 was increased 30 min post-high-frequency stimulation and blocked by N-methyl- d-aspartate receptor antagonists (AP-5 and MK-801). In sham group and controls, these changes were not observed. The expression of several other synaptic proteins (NR2B, glutamate receptors 2/3, N-ethylmaleimide sensitive factor) was not affected by long term potentiation induction. In hippocampal homogenates, the level of these proteins remained unchanged. These data indicate that N-methyl- d-aspartate receptor-dependent long term potentiation in CA1 region in vivo mainly affects the synaptic expression of glutamate receptor subunits and associated proteins in the whole hippocampus. The alteration of molecular aspects can play a role in regulating the long-lasting synaptic modification in hippocampal long term potentiation in vivo.