Abstract
We sought to determine whether the strict segregation of MAP2 and tau into somatodendritic and axonal compartments in situ was maintained in dissociated neuronal cultures of the rat cerebrum. Cultures grown under serum-free conditions were immunolabeled with monoclonal antibodies specific for MAP2 and tau. At 14 d after plating, a clear distinction between MAP2- and tau-immunoreactive neurites was apparent. MAP2- immunoreactive neurites were relatively short, thick, tapering, and branched. Tau-immunoreactive neurites formed a crisscrossing meshwork of long, fine-caliber neurites, which, in more densely plated cultures, had a tendency to form thick, ropelike fascicles. Unlike the MAP2 pattern, tau antibodies labeled somata only lightly. Since distinct populations of neurites were labeled with the 2 antibodies, we sought to observe the development of the topographically distinct compartments by double-labeled immunocytochemistry with both polyclonal and monoclonal antibodies to MAP2 and tau. Cells observed within the first 8 hr after plating demonstrated equally intense MAP2 and tau immunoreactivity in a coextensive distribution throughout the cell body and initial neurites. By 16 hr, some neurites began to assume dendritic and axonal features; however, many such processes contained reaction product for both MAP2 and tau. Beginning at this time, neurites that appeared axonal showed a progressively weaker reaction with MAP2 antibodies, and neurites that appeared dendritic showed a progressively weaker reaction with tau antibodies. In most neurites the diminution appeared to occur uniformly over the entire extent of the neurite. During this transformation period there were occasional axon-like neurites that contained MAP2 immunoreactivity proximally, while tau immunoreactivity extended over the entire length of the neurite. We conclude that neurons in culture are able to compartmentalize MAP2 and tau into their appropriate processes and only attain an apparently homogeneous population of one of these MAPs after the neuron has assumed dendritic and axonal features. The analysis also lends indirect support to the hypothesis that microtubule-associated proteins (MAPs) form this association at the distal extent of the growing neurite.
