GDNF gene delivery to injured adult CNS motor neurons promotes axonal growth, expression of the trophic neuropeptide CGRP, and cellular protection

Abstract

Glial-cell-line--derived neurotrophic factor (GDNF) has been identified as a potent survival and differentiation factor for several neuronal populations in the central nervous system (CNS), but to date, distinct effects of GDNF on motor axon growth and regeneration in the adult have not been demonstrated. In the present study, ex vivo gene delivery was used to directly examine whether GDNF can influence axonal growth, expression of neuronal regeneration-related genes, and sustain the motor neuronal phenotype after adult CNS injury. Adult Fischer 344 rats underwent unilateral transections of the hypoglossal nerve, followed by intramedullary grafts of fibroblasts genetically modified to secrete GDNF. Control animals received lesions and grafts of cells expressing a reporter gene. Two weeks later, GDNF gene delivery (1) robustly promoted the growth of lesioned hypoglossal motor axons, (2) altered the expression and intracellular trafficking of the growth-related protein calcitonin gene-related peptide (CGRP), and (3) significantly sustained the cholinergic phenotype in 84 +/- 6% of hypoglossal neurons compared with 39 +/- 6% in control animals (P < 0.001). This is the first neurotrophic factor identified to increase the in vivo expression of the trophic peptide CGRP and the first report that GDNF promotes motor axonal growth in vivo in the adult CNS. Taken together with previous in vitro studies, these findings serve as the foundation for a model wherein GDNF and CGRP interact in a paracrine manner to regulate neuromuscular development and regeneration.