Preferential expression of an AAV-2 construct in NOS-positive interneurons following intrastriatal injection

Most CNS studies using recombinant adeno-associated virus type 2 (rAAV-2) vectors have focused on gene delivery for the purpose of gene therapy. In the present study, we examined the feasibility of using rAAV-2 vectors to study the regulation of preprotachykinin-A (PPT-A) promoter activity in striatal medium spiny projection neurons. An rAAV-2 vector incorporating a PPT promoter fragment (shown previously to confer some cell-specificity of expression in vitro) coupled to a green fluorescent protein (GFP) reporter gene was stereotaxically injected into the rat striatum. Since medium spiny projection neurons represent the predominant neuronal type (90-95%) in the striatum, we predicted that the vast majority of GFP-expressing cells would be of this phenotype. Surprisingly, the transgene was actually expressed in a similar number of medium spiny projection neurons and interneurons, while glial expression of GFP was not observed. A preponderance of GFP-expressing interneurons was immunoreactive for the marker neuronal nitric oxide synthase (nNOS). Our results suggest that viral vector-related events that occur during transduction are the determining factor in the pattern of transgene expression observed, while the influence of the transgene promoter appears to be secondary, at least under the conditions employed.