Direct visualization of CaMKII at postsynaptic densities by electron microscopy tomography.

Ca(2+) /calmodulin-dependent protein kinase II (CaMKII) is a major component of postsynaptic densities (PSDs) involved in synaptic regulation. It has been previously shown that upon activity CaMKII from the spine reversibly aggregates at the cytoplasmic surfaces of PSDs, where it encounters various targets for phosphorylation. Targets for CaMKII are also present within the PSD, but there has been no reliable method to pinpoint whether, or where, CaMKII is located inside the PSD.

A negative stain for electron microscopic tomography.

While negative staining can provide detailed, two-dimensional images of biological structures, the potential of combining tomography with negative staining to provide three-dimensional views has yet to be fully realized. Basic requirements of a negative stain for tomography are that the density and atomic number of the stain are optimal, and that the stain does not degrade or rearrange with the intensive electron dose (~10⁶ e/nm²) needed to collect a full set of tomographic images. A commercially available, tungsten-based stain appears to satisfy these prerequisites.

Isolation and ultrastructural characterization of squid synaptic vesicles.

Synaptic vesicles contain a variety of proteins and lipids that mediate fusion with the pre-synaptic membrane. Although the structures of many synaptic vesicle proteins are known, an overall picture of how they are organized at the vesicle surface is lacking. In this paper, we describe a better method for the isolation of squid synaptic vesicles and characterize the results.

Two-stage surface freezing of a single top layer in a smectic-A membrane.

The crystallization of a single liquid top layer of smectic membranes of the compound 4O.8 has been studied with grazing-incidence x-ray diffraction. As this process takes place in two steps, involving an intermediate hexatic smectic-B layer before the final crystalline-B surface structure is reached, it provides a model for melting in two dimensions.

Smectic membranes in motion: approaching the fast limits of x-ray photon correlation spectroscopy.

The dynamics of the layer-displacement fluctuations in smectic membranes have been studied by x-ray photon correlation spectroscopy (XPCS). We report transitions from an oscillatory damping regime to simple exponential decay of the fluctuations, both as a function of membrane thickness and upon changing from specular to off-specular scattering. This behavior is in agreement with recent theories.