Inhibition of axonal transport 'in vivo' by a tubulin-specific antibody.

We have used antibodies against the major proteins of the cytoskeleton-tubulin, the neurofilament triplet proteins and actin-as in vivo probes to determine the contribution of separate components of the cytoskeleton in axonal transport. The injection of either Fast Blue or wheat germ agglutinin conjugated horseradish peroxidase into the caudate nucleus of adult rats resulted in the retrograde transport of these tracers to the neuronal cell bodies in the substantia nigra pars compacta. In experimental animals these tracer injections were immediately preceded by injections of antiserum against tubulin, neurofilament triplet protein or actin, into multiple sites in the caudate. Preimmune serum injection preceded tracer injection as a control in the contralateral caudate of the same animal. One antiserum against electrophoretically purified pig brain tubulin (NS-20) produced a dramatic decrease in the normal retrograde and anterograde transport of both tracers to the SN. Other antisera against tubulin, as well as neurofilament and actin antisera, had no effect on the axonal transport of the tracers. Affinity purified antibodies prepared from the NS-20 antitubulin serum also blocked axonal transport of the tracers. These results provide further support for a critical role of microtubules in axonal transport in vivo. Moreover, an antigenic determinant on tubulin that is uniquely recognized by the NS-20 antibodies may provide us with a way to define the site of association of transfer vesicles with microtubules.