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Journal of Neuroscience, Vol 4, 394-410, Copyright © 1984 by Society for Neuroscience
Differential subcellular localization of tubulin and the microtubule- associated protein MAP2 in brain tissue as revealed by immunocytochemistry with monoclonal hybridoma antibodies
A Caceres, LI Binder, MR Payne, P Bender, L Rebhun and O Steward
The distribution and subcellular localization of tubulin and MAP2 in brain
tissue were analyzed by immunocytochemistry with monoclonal hybridoma
antibodies prepared against Chinese hamster brain tubulin and MAP2. We
examined three anti-tubulin hybridoma antibodies (Tu3B, Tu9B, Tu12)
specific for beta-tubulin, and two anti-MAP2 hybridoma antibodies
(AP9,AP13). The specificity of each of the monoclonal antibodies was
characterized by staining nitrocellulose electrophoretic blots of SDS-
polyacrylamide gels of whole brain or hippocampal extracts. Each hybridoma
antibody bound only its respective antigen in these preparations.
Polyclonal antisera against tubulin were also examined. Sections reacted
with antisera against tubulin or monoclonal antibodies against beta-tubulin
revealed a wide variety of stained cellular compartments. The reaction
product was found to decorate dendritic and axonal microtubles in neurons;
glial cells were also stained. MAP2 immunoreactivity was found only in
neurons. In the case of one of the monoclonal antibodies (AP9), staining
was preferentially associated with dendritic processes. However, light but
significant staining of axonal processes was seen with AP13. Within
dendrites, MAP2 was found associated with dendritic microtubules and
postsynaptic densities (psd), both in shaft and spine synapses. In
addition, strong immunoreactivity for MAP2 was found within the cytoplasm
of dendritic spines. There was little or no immunoreactivity for tubulin in
the spine cytoplasm, although the psd was stained. The localization of MAP2
in dendritic spines and in the psd suggests that this protein may have a
biological role independent of its association with microtubules. The
observations on differential staining of the hybridoma antibodies against
MAP2 suggest that there may be distinct subtypes or states of MAP2 within
neurons.
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