INCREASED GABA (A) RECEPTOR [alpha]4 PROMOTER ACTIVITY FOLLOWING PILOCARPINE INDUCED STATUS-EPILEPTICUS IN ADULT RATS
Abstract number :
1.170
Submission category :
Year :
2003
Submission ID :
3724
Source :
www.aesnet.org
Presentation date :
12/6/2003 12:00:00 AM
Published date :
Dec 1, 2003, 06:00 AM
Authors :
Yogendra Sinh H. Raol, Margaret A. Maronski, John H. Wolfe, Shelly J. Russek, Amy R. Brooks-Kayal Neurology, Children[apos]s Hospital of Philadelphia, Philadelphia, PA; Pharmacology, Boston University School of Medicine, Boston, MA; Neurology and Pediatri
The adeno-associated virus (AAV) vector has been preferentially used in the central nervous system due to its predominantly neuronal transduction, higher stability and lack of any toxicity. Earlier studies have shown increased expression of [alpha]4 subunit and decreased [alpha]1 subunit in dentate granule cells following pilocarpine-induced status-epilepticus (SE) in adult rats. In the present study we explore the possibility of using AAV 2 vector as a genetic tool to examine the regulation of GABA[sub]A[/sub] receptor subunits in dentate gyrus following SE in adult rats.
Adult male Sprague-Dawley rats were injected with 2 [mu]l of AAV 2 virus (5.2x1012 genomic particles/ml) containing an [alpha]4 promoter driving the expression of eYFP reporter gene into upper and lower blades of dentate gyrus. Two weeks following viral injections, rats were injected with pilocarpine to induce SE. The control rats were injected with 1/10th of the convulsive dose. One and two weeks following SE, rats were perfused with 4% paraformaldehyde. Non-radioactive in situ hybridization was used to detect the expression of eYFP reporter mRNA.
An increase in [alpha]4 promoter driving reporter gene expression was found both one and two weeks after pilocarpine induced SE compared to control animals.
We show increased expression of eYFP reporter gene driven by [alpha]4 promoter following SE suggesting that previously reported increase in [alpha]4 subunit following SE are occurring at a transcriptional level. This system may be useful for modifying gene expression in dentate granule cells following SE.
[Supported by: NIH NS42363-01 to ABK and SJR]