Cytoskeletal Structures and Oligodendroglial Differentiation in C-6 Glial Cells

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Abstract
The relationship of the cytoskeleton to a biochemical expression of oligodendroglial differentiation was studied in cultured [rat] C-6 glial cells. The effect of the cytoskeletal perturbants, colchicine and cytochalasin D, on the induction of the oligodendroglial marker enzyme, 2'',3''-cyclic nucleotide 3''-phosphohydrolase (CNP), caused by removal of serum from the culture medium was studied. Each drug inhibited CNP induction in a concentration-dependent manner, and essentially complete inhibition of induction was observed with 0.25 .mu.M colchicine or 2.0 .mu.M cytochalasin D. Detailed study of the effect of colchicine was carried out. This antimicrotubular agent not only totally prevented induction if added at the onset of serum removal, but also prevented further induction when added at various times after serum removal. That the effect of colchicine related to the drug''s effect on microtubules was supported by the demonstration that lumicolchicine, a colchicine isomer which has no effect on microtubules, had no effect on the CNP induction. Colchicine, but not lumicolchicine, prevented the morphological signs of differentiation provoked by serum removal. The effect of colchicine was reversible and relatively specific. No concomitant effect of colchicine on the activity of another plasma membrane enzyme of C-6 cells, i.e., (Na+ + K+)-activated ATPase, or on the rate of incorporation of [3H]leucine into total protein of intact cells could be discerned. The possibility that the site of the effect of colchicine is on intracellular events was suggested by the observation that the drug inhibited the induction of CNP by dibutyryl cAMP. Probably the cytoskeleton is involved in oligodendroglial differentiation.