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Journal of Neuroscience, Vol 7, 3142-3153, Copyright © 1987 by Society for Neuroscience
MAP2 and tau segregate into dendritic and axonal domains after the elaboration of morphologically distinct neurites: an immunocytochemical study of cultured rat cerebrum
KS Kosik and EA Finch
Department of Medicine, Brigham and Women's Hospital, Boston, Massachusetts 02115.
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.
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