title: Subcellular organization of camkii in rat hippocampal pyramidal neurons.
abstract: Calcium/calmodulin-dependent protein kinase II (CaMKII) plays a key role in N-methyl-D-aspartate (NMDA) receptor-dependent long-term synaptic plasticity; its location is critical for signal transduction, and may provide clues that further elucidate its function. We therefore examined the subcellular localization of CaMKII in CA1 stratum radiatum of adult rat hippocampus, by using immuno-electron microscopy after chemical fixation. When tissue was fixed quickly, the concentration of CaMKIIα (assessed by pre-embedding immunogold) was significantly higher in dendritic shafts than in spine heads. However, when tissue was fixed 5 minutes after perfusion with normal saline, the density of labeling decreased in dendritic shaft while increasing in spine heads, implying rapid translocation into the spine during brief perimortem stress. Likewise, in quickly fixed tissue, CaMKII within spine heads was found at comparable concentrations in the "proximal" half (adjacent to the spine neck) and the "distal" half (containing the postsynaptic density [PSD]), whereas after delayed fixation, label density increased in the distal side of the spine head, suggesting that CaMKII within the spine head moves toward the PSD during this interval. To estimate its distribution at the synapse in vivo, we performed postembedding immunogold staining for CaMKII in quick-fixed tissue, and found that the enzyme did not concentrate primarily within the central matrix of the PSD. Instead, labeling density peaked ∼40 nm inside the postsynaptic membrane, at the cytoplasmic fringe of the PSD. Labeling within 25 nm of the postsynaptic membrane concentrated at the lateral edge of the synapse. This lateral "PSD core" pool of CaMKII may play a special role in synaptic plasticity.
authors.affiliation: Department of Ophthalmology, Duke University Medical Center, Durham, North Carolina, 27710.
authors.forename: Jin-Dong
authors.initials: JD
authors.lastname: Ding
authors.forename: Mary B
authors.initials: MB
authors.lastname: Kennedy
authors.forename: Richard J
authors.initials: RJ
authors.lastname: Weinberg
mesh_terms: D000818:Animals; D000906:Antibodies / Q000737:chemistry; D054732:Calcium-Calmodulin-Dependent Protein Kinase Type 2 / Q000378:metabolism*; D002462:Cell Membrane / Q000201:enzymology; D003593:Cytoplasm / Q000201:enzymology; D003627:Data Interpretation, Statistical; D003712:Dendrites / Q000201:enzymology; D049229:Dendritic Spines / Q000201:enzymology; D006624:Hippocampus / Q000166:cytology / Q000201:enzymology*; D007091:Image Processing, Computer-Assisted; D007150:Immunohistochemistry; D016253:Microscopy, Immunoelectron; D017966:Pyramidal Cells / Q000201:enzymology*; D051381:Rats; D017207:Rats, Sprague-Dawley; D013347:Subcellular Fractions / Q000378:metabolism*; D013569:Synapses / Q000201:enzymology; D016610:Tissue Embedding; D016707:Tissue Fixation
chemical_list: D000906:Antibodies; D054732:Calcium-Calmodulin-Dependent Protein Kinase Type 2
keywords: hypoxia; immunogold electron microscopy; postsynaptic density; synaptic plasticity