NCAM180 and glutamate receptor subtypes in potentiated spine synapses: an immunogold electron microscopic study.

Activity-dependent changes in expression and localization of the largest major isoform of the neural cell adhesion molecule NCAM180 and three subtypes of glutamate receptors predominantly expressed in the outer part of the molecular layer of the dentate gyrus of adult rats-the NMDA receptor NR2A, the AMPA receptor GluR2/3, and the metabotropic glutamate receptor mGluR1 - were investigated using postembedding immunogold labeling, and electron microscopy. In synaptic membranes of nonstimulated spine synapses, NCAM180 and NR2A accumulated in the center of the postsynaptic density, whereas GluR2/3 and mGluR1 were distributed evenly. Twenty-four hours following induction of long-term potentiation in vivo, NCAM180 and NR2A accumulated at the edges of postsynaptic densities, whereas GluR2/3 was localized more centrally. Also, the distribution of gold particles per synapse significantly changed for NCAM180, NR2A, and mGluR1. Thus, changes in synaptic strength are associated with concomitant changes in the expression and distribution of NCAM180 and glutamate receptors, particularly of the NR2A subtype.

Persistent phosphorylation of cyclic AMP responsive element-binding protein and activating transcription factor-2 transcription factors following transient cerebral ischemia in rat brain.

The transcription factors cyclic AMP responsive element-binding protein (CREB) and activating transcription factor-2 were studied in rat brains subjected to 15 min ischemia followed by varied periods of reperfusion using western blot and immunocytochemical analyses. The total amounts of both CREB and activating transcription factor-2 were not altered in the hippocampus after ischemia. In contrast, levels of the phosphorylated forms of both transcription factors decreased during ischemia but rebounded following reperfusion. The phospho-forms of CREB and activating transcription factor-2 showed regional and temporal differences in their expression. Phospho-CREB was increased relative to control levels at 30 min, and continued to increase for at least three days postischemia, mainly in dentate granule cells. The level of phospho-activating transcription factor-2 appeared to be higher in CAI pyramidal cells than in dentate granule cells after ischemia. The present findings suggest that the signaling pathways for phosphorylation of CREB may be neuroprotective for dentate cells, which are relatively resistant to ischemic insults. The increased phospho-activating transcription factor-2 may reflect increased stresses in these neurons. The more modest activation of CREB pathways in CA1 neurons may not be enough to overcome the increased stresses in these neurons, contributing to delayed neuronal death.