DNA plasmid that codes for human Bcl-2 gene preserves axotomized Clarke's nucleus neurons and reduces atrophy after spinal cord hemisection in adult rats

• Published in: Journal of comparative neurology (1911) 1999-02, Vol.404 (2), p.159-171
• Main Authors: Takahashi, Kosei, Schwarz, Emily, Ljubetic, Cecilia, Murray, Marion, Tessler, Alan, Saavedra, Raul A.
• Format: Article
• Language: English
• Subjects: Animals; apoptosis; Atrophy; Axotomy; Cell Survival - physiology; Denervation; DNA - genetics; DNA vector; Female; Fluorescent Dyes; Gene Expression - physiology; Genes, bcl-2 - genetics; Genes, Reporter - genetics; Humans; Lac Operon - genetics; neuronal rescue; Neurons - physiology; Plasmids - genetics; Plasmids - physiology; Rats; Rats, Sprague-Dawley; Spinal Cord - pathology; Stilbamidines
• ISSN: 0021-9967; 1096-9861
• DOI: 10.1002/(SICI)1096-9861(19990208)404:2<159::AID-CNE2>3.0.CO;2-E

Spinal cord injury in adult mammals causes atrophy or death of some axotomized neurons. The product of the antiapoptotic gene Bcl‐2 prevents neuron death in vivo. We delivered Bcl‐2 by intraspinal injection of a DNA plasmid encoding this gene to determine if axotomized neurons destined to undergo retrograde death could be rescued. Axons of the right side Clarke's Nucleus (CN) were cut unilaterally in adult Sprague‐Dawley rats by T8 hemisection, leaving the contralateral (left) CN as an intact control. Two months postoperatively, there was ∼35% loss of total CN neurons in the right L1 segment. Only 15% of large CN neurons (>400 μm2), whose axons project to the cerebellum, survived—indicating atrophy and/or death of 85% of these cells. We injected a DNA plasmid encoding the human Bcl‐2 gene and the bacterial reporter gene LacZ, which was complexed with cationic lipids, into the right side of segment T8 of the normal spinal cord, or just caudal to the hemisection site. The reporter gene was expressed in the perikarya of right CN neurons at L1 for up to 7 days, but not 14 days. Two months following T8 hemisection and Bcl‐2/LacZ DNA injection, there was no significant loss of CN neurons ipsilateral to the lesion. Surprisingly, 61% of large neurons survived, indicating partial protection from atrophy. In contrast, a DNA plasmid that codes for the LacZ reporter gene, but not Bcl‐2, did not prevent CN neuron death or atrophy. Administration of the Bcl‐2 gene in adult rats and its expression in these CNS neurons prevents retrograde cell death, and also minimizes atrophy. These results may serve as the basis for developing novel gene therapy strategies for patients with spinal cord injury. J. Comp. Neurol. 404:159–171, 1999. © 1999 Wiley‐Liss, Inc.