Alzheimer's disease amyloid precursor protein on the surface of cortical neurons in primary culture co-localizes with adhesion patch components

Abstract

Immunofluorescence on primary dissociated rat neuronal cultures (cortical, hippocampal, and cerebellar) and organotypic hippocampal cultures was used to investigate the pattern of distribution of cell-surface amyloid precursor protein (APP). Antibodies directed against the extracellular (N-terminal) portion of APP or against the entire molecule, but not against the C-terminal portion, revealed a striking segmental pattern of immunoreactivity along both axons and dendrites of all neuronal types tested. The pattern first developed between 24 and 48 h in culture. The segments showed co-localization with β 1-integrin and talin immunoreactivities, but not with GAP-43 or clathrin, indicating that they may mark adhesion patches. Confocal laser microscopy supported a surface location for the APP responsible for the segmental pattern on neurites, as did the reduction of segmental immunoreactivity after exposure to μ-calpain or trypsin. It is conjectured that APP may have a role in cell-substratum interactions in the medium term, during such events as synaptic plasticity and neurite stability during extension.