Cytophotometric quantification of retrograde axonal transport of a fluorescent tracer (primuline) in mouse facial neurons

Abstract

A method for cytophotometric quantifications of retrograde axonal transport of a fluorescent tracer in tissue sections is described. As a fluorescent tracer the anionic vital stain primuline proved to be suitable since it resulted in a strong yellow-green fluorescence, which faded very slowly permitting localization of cells during illumination with UV-light. Primuline injected into the muscles of the vibrissae in mice was transported to the corresponding nerve cell bodies in the facial nucleus, where it appeared as fluorescent granules 9 h after the injection. The fluorescence intensity increased with increasing exposure times and concentrations of the injected tracers. The motor endplates showed no ultrastructural changes after the tracer injections. The motor endplates showed no ultrastructural changes after the tracer injections. With this method a substantial increase in tracer accumulation in facial neurons could be revealed during nerve regeneration 11 days after crushing the facial nerve. Small alterations in neuronal tracer accumulation could be measured after intoxication of the mice with botulinum and tetanus toxins. Since these toxins should cause a decrease or increase in the degree of synaptic activity the amount of retrograde axonal transport may to a certain extent be dependent on the synaptic function. The findings with this new technique therefore indicate that quantitative changes occur in axonal transport in materials from the periphery during different pathological and physiological conditions, which may be important for an understanding of how a nerve cell body is dependent on its peripheral field of innervation.