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Volume 17, Number 12, Issue of June 15, 1997 pp. 4623-4632
Copyright ©1997 Society for Neuroscience

Expression of Specific Tubulin Isotypes Increases during Regeneration of Injured CNS Neurons, But Not after the Application of Brain-Derived Neurotrophic Factor (BDNF)

Received Dec. 18, 1996; revised Feb. 14, 1997; accepted April 2, 1997.

Alyson E. Fournier and Lisa McKerracher

Faculté de Médecine, Département de Pathologie, Université de Montréal, and McGill University, C.P. 6128, Succursale Centre-ville Montréal, Québec H3C 3J7, Canada

Axonal regrowth after injury is accompanied by changes in the expression of tubulin, but the contributions of substrate molecules and neurotrophic factors in regulating these changes in vivo are not known. Adult rat retinal ganglion cells (RGCs) were examined after intraorbital axotomy, after application of a peripheral nerve (PN) graft to stimulate regeneration, and after axotomy and treatment with brain-derived neurotrophic factor (BDNF). After these treatments we used in situ hybridization to study mRNA levels for I, II, III, IV_a, and T1 tubulin isotypes. Levels of mRNA for all isotypes were downregulated after intraorbital axotomy. During regrowth of injured RGC axons, mRNA levels for II, III, and T1 isotypes were upregulated specifically and dramatically, suggesting that elevated expression of these isotypes is correlated specifically with axonal regrowth. A corresponding increase in III protein levels was detected by immunocytochemistry. The I and IV_a mRNAs were not increased during regeneration. BDNF did not elicit a specific increase in the mRNA levels for the III and T1 isotypes and had only a small effect on mRNA levels for the II isotype. Therefore, despite the ability of BDNF to support the survival of injured RGCs and to enhance neurite outgrowth of retinal neurons in vitro, the in vivo application of BDNF alone is unable to induce the program of changes in growth-associated tubulins that accompany regeneration of RGC axons into PN grafts. We speculate that, in addition to BDNF, cooperative signaling with substrate molecules is required to allow RGCs to regenerate and exhibit tubulin isotype switching.

Key words: BDNF; tubulin isotypes; microtubules; rat; regeneration; axotomy

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