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Published March 1973 | Published
Journal Article Open

The Effect of Vincristine Sulphate on the Axoplasmic Flow of Proteins in Cultured Sympathetic Neurons

Abstract

The effect of vincristine sulphate on the axoplasmic flow of labelled proteins in neurites of chick embryo sympathetic neurons growing in tissue culture was studied by autoradiography. In control neurons most of the 3H-proteins synthesized during a 90-min pulse with a 3H-amino acid were localized in cell bodies. There was a diminishing gradient of labelled proteins in the neurites which was highest in portions adjacent to the cell bodies and lowest at the periphery. During a physiological chase there was a gradual increase in the amount of label in the neurites, so that after a 15-h chase even the most peripheral portions were well labelled. This indicates that a portion of the labelled proteins synthesized in the cell bodies are transported peripherally into the neurites. The centrifugal movement of labelled proteins in neurites was markedly decreased when cells were grown in medium containing 10 µg/ml vincristine sulphate. After a 15-h chase in the presence of drug only a small amount of label was in the peripheral portion of the neurites. Treatment with vincristine did not decrease the rate of amino acid incorporation or alter the rate of protein turnover during the course of the experiment. Thus an explanation of the results based on an altered rate of total cell protein synthesis or degradation is unlikely. The capacity of sympathetic neurons to take up and concentrate exogenous [3H]norepinephrine in their neurites was only slightly reduced by vincristine. This indicates that at least some cellular activities requiring metabolic energy are relatively unaffected by the interruption in axoplasmic flow caused by vincristine and that the mechanism by which vincristine interferes with axoplasmic flow does not involve general cellular toxicity. The major morphological differences between control and vincristine-treated neurons were the absence of microtubules and the presence of crystal-like structures within the cells. The relationship between the effect of vincristine on the axoplasmic flow of proteins and the arrangement of the microtubule system is discussed.

Additional Information

Copyright © 1973 by Company of Biologists. Submitted on July 17, 1972. This work was supported by a research grant from the U.S. Public Health Service (CA-10755)to M.N.G. and by a research fellowship from the National Institute of General Medical Sciences (4-F01-GM-41039) to J.M.E.

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