Neurite elongation is blocked if microtubule polymerization is inhibited in PC12 cells.
We have injected process-bearing PC12 cells with colchicine-tubulin mixed with either fluorescein-dextran or a rhodamine-labelled tubulin analogue to determine the role of microtubule polymerization in neurite elongation. Colchicine-tubulin is a specific, substoichiometric poison of microtubule assembly. We have shown that colchicine-tubulin does not cause existing PC12 microtubules to disassemble, and yet can inhibit the assembly of rhodamine-tubulin injected along with it. In population studies of neurite outgrowth in injected and uninjected cells, we find that colchicine-tubulin substantially inhibits neurite extension from injected cells over a wide variety of concentrations. In acute time-course studies of injected cells, we find that colchicine-tubulin does not block neurite outgrowth until the injectate reaches the neurite tip. Thereafter, however, it blocks process elongation completely. Thus we can conclude that microtubule polymerization in the region of the growth cone is an important element in neurite elongation. While polymerization at the cell body may be important in supplying subunits to the distal neurite, it does not play a direct role in process extension.
Keith
C H
CH
Department of Zoology, University of Georgia, Athens 30602.
D000818:Animals; D001369:Axons / Q000187:drug effects / Q000502:physiology*; D002460:Cell Line; D003078:Colchicine / Q000494:pharmacology; D005455:Fluorescent Antibody Technique; D005456:Fluorescent Dyes; D008856:Microscopy, Fluorescence; D008870:Microtubules / Q000187:drug effects* / Q000378:metabolism / Q000502:physiology*; D012235:Rhodamines; D014404:Tubulin / Q000378:metabolism / Q000494:pharmacology
D005456:Fluorescent Dyes; D012235:Rhodamines; D014404:Tubulin; D003078:Colchicine