Abstract
Changes in the amounts of tubulin, actin, and neurofilament polypeptides were found in regenerating motoneurons of grass frogs during the period of axonal elongation. Ventral roots 9 and 10 were transected unilaterally about 7 mm from the spinal cord. 35 d later, [3H]colchicine binding had decreased in the proximal stumps to approximately one-half of contralateral control values, well before the regenerating motor axons had reinnervated skeletal muscles of the hind limb. [3H]colchicine binding did not change significantly in the operated halves of the 9th and 10th spinal cord segments over a 75-d period. The relative amounts of actin, tubulin, and neurofilament polypeptides in the operated ventral roots were measured by quantitative densitometry of stained two-dimensional electrophoretic gels. Alpha-tubulin, beta-tubulin, and the 68,000 molecular weight subunit of neurofilaments (NF68) decreased within the transected ventral roots to 78%, 57%, and less than 15% of control values, respectively. The amount of actin increased to 132% of control values within the operated ventral roots, although this change was not statistically significant. Opposite changes were found within motoneuronal cell bodies isolated from the spinal cord. The relative amounts of alpha-tubulin, beta-tubulin and NF68 within axotomized perikarya increased, respectively, to 191%, 146%, and 144% of that in control perikarya isolated from the contralateral side of the spinal cord. Thus, the changes in NF68 and tubulin did not occur uniformly throughout the injured cells. The possible structural and functional consequences of these changes are discussed.
Highlights
Changes in the amounts of tubulin, actin, and neurofilament polypeptides were found in regenerating motoneurons of grass frogs during the period of axonal elongation
The binding of [aH]colchicine by tubulin dimers was assayed in homogenates of spinal cord and ventral roots between 1 and 75 d after ventral root transection (Fig. 3)
Tubulin and NF68 increased within the axotomized cell bodies and decreased within the proximal stumps of the transected ventral roots, whereas actin increased within the roots
Summary
Changes in the amounts of tubulin, actin, and neurofilament polypeptides were found in regenerating motoneurons of grass frogs during the period of axonal elongation. The relative amounts of alpha-tubulin, beta-tubulin and NF68 within axotomized perikarya increased, respectively, to 191%, 146%, and 144%of that in control perikarya isolated from the contralateral side of the spinal cord. Increased amounts of radioactivity associated with newly synthesized tubulin and actin are transported within regenerating optic [17, 56] and motor axons [30, 34] following administration of labeled amino acids to their cell bodies. We looked for changes in the total amounts of individual cytoskeletal proteins within the perikarya and axons of axotomized spinal motoneurons of the frog. The cell bodies of axotomized motoneurons, obtained by a new procedure for isolating perikarya from small neuronal populations, and the proximal stumps of transected ventral roots
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