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

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 Previous Article

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

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




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