BDNF AND NT-4 5 PREVENT ATROPHY OF RAT RUBROSPINAL NEURONS AFTER CERVICAL AXOTOMY, STIMULATE GAP-43 AND T-ALPHA-1-TUBULIN MESSENGER-RNA EXPRESSION, AND PROMOTE AXONAL REGENERATION/

Rubrospinal neurons (RSNs) undergo a marked atrophy in the second week after cervical axotomy. This delayed atrophy is accompanied by a decl ine in the expression of regeneration-associated genes such as GAP-43 and T alpha 1-tubulin, which are initially elevated after injury. Thes e responses may reflect a deficiency in the trophic support of axotomi zed RSNs. To test this hypothesis, we first analyzed the expression of mRNAs encoding the trk family of neurotrophin receptors. In situ hybr idization revealed expression of full-length trkB receptors in virtual ly all RSNs, which declined 7 d after axotomy. Full-length trkC mRNA w as expressed at low levels. Using RT-PCR, we found that mRNAs encoding trkC isoforms with kinase domain inserts were present at levels compa rable to that for the unmodified receptor. TrkA mRNA expression was no t detected in RSNs, and the expression of p75 was restricted to a smal l subpopulation of axotomized cells. in agreement with the pattern of trk receptor expression, infusion of recombinant human BDNF or NT-4/5 into the vicinity of the axotomized RSNs, between days 7 and 14 after axotomy, fully prevented their atrophy. This effect was still evident 2 weeks after the termination of BDNF treatment. Moreover, BDNF or NT- 4/5 treatment stimulated the expression of GAP-43 and T alpha 1-tubuli n mRNA and maintained the level of trkB expression. Vehicle, NGF, or N T-3 treatment had no significant effect on cell size or GAP-43 and T a lpha 1-tubulin expression. In a separate experiment, infusion of BDNF also was found to increase the number of axotomized RSNs that regenera ted into a peripheral nerve graft. Thus, in BDNF-treated animals, the prevention of neuronal atrophy and the stimulation GAP-43 and T alpha 1-tubulin expression is correlated with an increased regenerative capa city of axotomized RSNs.