Training in chicks alters PSA-N-CAM distribution in forebrain cell membranes.

The intermediate and medial hyperstriatum ventrale (IMHV) in the chick is involved in memory formation following one-trial passive avoidance training. Possible links between neural cell adhesion molecule (N-CAM) distribution and memory consolidation were examined in an immunoelectron microscope study of IMHV 6 h after training. An antibody against alpha 2,8 polysialic acid (PSA), characteristic for embryonic N-CAM, and one against the protein backbones of N-CAM were labelled (post-embedding), separately, with 15 nm immunogold, and binding to their epitopes was analysed using the statistics of point processes. No difference in labelling levels between control and trained chicks, for either antibodies, was found, both groups of birds showing that cell membrane regions are 10-30 times more enriched in N-CAM isoforms, and have a two-fold greater proportion of PSA-N-CAM, than tissue as a whole. However, 375-400 nm regular arrays of labelled PSA-N-CAM were revealed statistically in cell membranes of control, but not trained, chicks, which may be related to the possible involvement of such membrane PSA domains in memory-related neuroplasticity.

Lateral patterns of the neural cell adhesion molecule on the surface of hippocampal cells developing in vitro.

In monolayer cultures of hippocampal neurons from newborn rats, an immunocytochemical quantitative study was carried out to investigate age-dependent arrangement of the neural cell adhesion molecules in different parts of cell membranes. On the fifth and 12th day in vitro, neural cell adhesion molecules were labelled with specific antibodies and protein A conjugated to colloidal gold particles. Samples of randomly selected electron micrographs that displayed labelled membrane fragments of cell bodies, growth cones, and axons were numerically analysed for the five- and 12-day in vitro neurons. Neural cell adhesion molecules surface topography was quantitatively described and compared, using a statistical stereological approach. The mean surface density of labelled neural cell adhesion molecules was found to be approximately 2.5 times higher in growth cone membranes relative to somatic and axonal membranes in five-day in vitro neurons. By the 12th day in vitro, this density decreases in somatic membranes (approximately 18%) and increases in axonal membranes (approximately 60%). Representative spectra of lateral intervals between labels as well as images that show typical topography of label on membrane surfaces were simulated. The results revealed regular patterns of neural cell adhesion molecules on the somatic surface and allowed consideration of neural cell adhesion molecules arrangement in a view of membrane adhesion properties. Participation of cytoskeleton in neural cell adhesion molecules rearrangement is discussed.

Changes in the neural cell adhesion molecule patterns on the rat glial cell surfaces with development and contact formation in vitro.

In monolayer cultures of newborn rat hippocampal cells, immunogold-labelling at the electron microscope level was used to study quantitatively the neural cell adhesion molecule (N-CAM) arrangement on the surface of glial soma and processes on 5 and 12 days in vitro (DIV). Four corresponding samples of micrographs were formed. To quantify the labelling, a stochastic geometry approach was used. Spectra of lateral distances between labels as well as simulated images of the surface label arrangement (invisible in micrographs) were derived and compared. The data show that, on both 5 and 12 DIV, N-CAM density on the surface of processes is approximately 2 times higher than that in somata; 12-DIV cells showing a lower (approximately 25%) N-CAM surface density as compared with the 5-DIV cells. This suggests that N-CAM expression in glia surfaces decreases while the cells form contacts, and N-CAM sorting between soma and processes remains stable. The simulated topographies of the lateral N-CAM arrangement might highlight fundamental mechanisms that underlie formation of the neural network.