Adeno-associated virus-mediated expression and constitutive secretion of galanin suppresses limbic seizure activity in vivo.
Intractable temporal lobe epilepsy presents an ideal target for gene therapy, but therapeutic success depends upon the ability to suppress limbic seizure activity. Adeno-associated virus vectors (AAV) were constructed in which the fibronectin secretory signal sequence (FIB) preceded the coding sequence for galanin (AAV-FIB-GAL) or green fluorescent protein (AAV-FIB-GFP), constructs that express and constitutively secrete the gene product. Bilateral AAV-FIB-GAL infusion into the rat piriform cortex (2 microl/side) significantly attenuated kainic acid-induced seizures (10 mg/kg, ip) such that 11/12 rats exhibited no limbic seizures, while the remaining rat exhibited only a brief, single class III seizure. This AAV-FIB-GAL infusion also prevented electrographic seizure activity. In contrast, bilateral AAV-FIB-GFP infusion did not alter either behavioral or electrographic seizure activity. Since prior seizure exposure could influence vector efficacy, another group of rats received daily electrical stimulation of the piriform cortex until three consecutive class V seizures were elicited. Subsequently, AAV-FIB-GAL or AAV-FIB-GFP (3 microl/30 min) was infused into the area of the electrode. One week later the AAV-FIB-GAL rats exhibited a significant increase in the stimulation current necessary to evoke limbic seizure activity, while AAV-FIB-GFP did not alter the seizure threshold. Thus, AAV-mediated galanin expression and secretion significantly suppress limbic seizure activity in vivo.
McCown
Thomas J
TJ
Gene Therapy Center and Department of Psychiatry, University of North Carolina at Chapel Hill School of Medicine, Chapel Hill, NC 27599, USA. Thomas_McCown@med.unc.edu
D000818:Animals; D002540:Cerebral Cortex / Q000187:drug effects / Q000378:metabolism / Q000503:physiopathology; D000229:Dependovirus / Q000235:genetics*; D004569:Electroencephalography / Q000379:methods; D005353:Fibronectins / Q000235:genetics / Q000378:metabolism; D019004:Galanin / Q000235:genetics / Q000378:metabolism*; D015870:Gene Expression / Q000235:genetics*; D015316:Genetic Therapy / Q000379:methods; D005822:Genetic Vectors / Q000008:administration & dosage / Q000235:genetics; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D007260:Infusion Pumps; D007608:Kainic Acid / Q000378:metabolism / Q000494:pharmacology; D008297:Male; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D051381:Rats; D017207:Rats, Sprague-Dawley; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012640:Seizures / Q000235:genetics / Q000503:physiopathology / Q000628:therapy*
D005353:Fibronectins; D012333:RNA, Messenger; D049452:Green Fluorescent Proteins; D019004:Galanin; D007608:Kainic Acid