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The Journal of Neuroscience, December 3, 2003, 23(35):11270-11278

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Cellular/Molecular
Distribution of Postsynaptic Density (PSD)-95 and Ca²⁺/Calmodulin-Dependent Protein Kinase II at the PSD

Jennifer D. Petersen,¹ Xiaobing Chen,¹ Lucia Vinade,¹ Ayse Dosemeci,² John E. Lisman,³ and Thomas S. Reese¹

¹Laboratory of Neurobiology, National Institute of Neurological Disorders and Stroke, National Institutes of Health, Bethesda, Maryland 20892, ²Program in Molecular Physiology, Marine Biological Laboratory, Woods Hole, Massachusetts 02543, and ³Volen Center for Complex Systems, Brandeis University, Waltham, Massachusetts 02454

Postsynaptic densities (PSDs) contain proteins that regulate synaptic transmission. We determined the positions of calcium/calmodulin-dependent protein kinase II (CaMKII) and PSD-95 within the three-dimensional structure of isolated PSDs using immunogold labeling, rotary shadowing, and electron microscopic tomography. The results show that all PSDs contain a central mesh immediately underlying the postsynaptic membrane. Label for PSD-95 is found on both the cytoplasmic and cleft sides of this mesh, averaging 12 nm from the cleft side. All PSDs label for PSD-95. The properties of CaMKII labeling are quite different. Label is virtually absent on the cleft sides of PSDs, but can be heavy on the cytoplasmic side at a mean distance of 25 nm from the cleft. In tomograms, CaMKII holoenzymes can be visualized directly, appearing as labeled, tower-like structures reflecting the 20 nm diameter of the holoenzyme. These towers protrude from the cytoplasmic side of the central mesh. There appears to be a local organization of CaMKII, as judged by fact that the nearest-neighbor distances are nearly invariant over a wide range of labeling density for CaMKII. The average density of CaMKII holoenzymes is highly variable, ranging from zero to values approaching a tightly packed state. This variability is significantly higher than that for PSD-95 and is consistent with an information storage role for CaMKII.

Key words: postsynaptic density; CaMKII; PSD; PSD-95; electron microscopy; tomography

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Received June 6, 2003; revised September 3, 2003; accepted October 8, 2003.

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