Functional characterization of NGF-secreting cell grafts to the acutely injured spinal cord.
Previously we reported that grafts of cells genetically modified to produce human nerve growth factor (hNGF) promoted specific and robust sprouting of spinal sensory, motor, and noradrenergic axons. In the present study we extend these investigations to assess NGF effects on corticospinal motor axons and on functional outcomes after spinal cord injury. Fibroblasts from adult rats were transduced to express human NGF; control cells were not genetically modified. Fibroblasts were then grafted to sites of midthoracic spinal cord dorsal hemisection lesions. Three months later, recipients of NGF-secreting grafts showed deficits on conditioned locomotion over a wire mesh that did not differ in extent from control-lesioned animals. On histological examination, NGF-secreting grafts elicited specific sprouting from spinal primary sensory afferent axons, local motor axons, and putative cerulospinal axons as previously reported, but no specific responses from corticospinal axons. Axons responding to NGF robustly penetrated the grafts but did not exit the grafts to extend to normal innervation territories distal to grafts. Grafted cells continued to express NGF protein through the experimental period of the study. These findings indicate that 1) spinal cord axons show directionally sensitive growth responses to neurotrophic factors, 2) growth of axons responding to a neurotrophic factor beyond an injury site and back to their natural target regions will likely require delivery of concentration gradients of neurotrophic factors toward the target, 3) corticospinal axons do not grow toward a cellular source of NGF, and 4) functional impairments are not improved by strictly local sprouting response of nonmotor systems.
Tuszynski
M H
MH
Department of Neurosciences, University of California-San Diego, La Jolla 92093-0608, USA.
Murai
K
K
Blesch
A
A
Grill
R
R
Miller
I
I
D000208:Acute Disease; D000818:Animals; D001369:Axons / Q000502:physiology; D015152:Blotting, Northern; D015740:Calcitonin Gene-Related Peptide / Q000032:analysis; D002478:Cells, Cultured; D002795:Choline O-Acetyltransferase / Q000032:analysis; D004299:Dopamine beta-Hydroxylase / Q000032:analysis; D005347:Fibroblasts / Q000166:cytology / Q000378:metabolism / Q000637:transplantation*; D015870:Gene Expression / Q000502:physiology; D018014:Gene Transfer Techniques; D006801:Humans; D008124:Locomotion / Q000502:physiology; D015335:Molecular Probes; D009414:Nerve Growth Factors / Q000235:genetics / Q000378:metabolism*; D009474:Neurons / Q000737:chemistry / Q000201:enzymology; D011712:Pyramidal Tracts / Q000166:cytology / Q000601:surgery; D012333:RNA, Messenger / Q000032:analysis; D051381:Rats; D011916:Rats, Inbred F344; D013119:Spinal Cord Injuries / Q000601:surgery*; D014446:Tyrosine 3-Monooxygenase / Q000032:analysis; D018998:Wheat Germ Agglutinin-Horseradish Peroxidase Conjugate
D015335:Molecular Probes; D009414:Nerve Growth Factors; D012333:RNA, Messenger; D018998:Wheat Germ Agglutinin-Horseradish Peroxidase Conjugate; D014446:Tyrosine 3-Monooxygenase; D004299:Dopamine beta-Hydroxylase; D002795:Choline O-Acetyltransferase; D015740:Calcitonin Gene-Related Peptide