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Transgene expression in neurons of rat nucleus tractus solitarii and pterygopalatine ganglion using adeno‐associated viral vectors

Adeno‐associated viral vector (AAV) has distinct advantages over other viral vectors in delivering genes of interest. AAV transfects mainly neurons, produces no toxicity or inflammatory responses and yields long‐term transgene expression. Previously we showed that adenovirus (Ad) mainly transfects glial cells and feline immunodeficiency virus (FIV) mainly transfects neurons in the nucleus tractus solitarii (NTS). However, gene transduction in the pterygopalatine ganglion (PPG) using either Ad or FIV has been unsatisfactory. In this study, we tested the hypothesis that AAV serotype 2 (AAV2) selectively transfects NTS and PPG neurons. Using double‐label fluorescent immunostaining combined with confocal microscopy we examined expression of the reporter gene enhanced green fluorescent protein (eGFP) after microinjecting AAV2eGFP into rat NTS or PPG. Robust expression of eGFP was observed in many cells of the NTS and PPG after introduction of AAV2eGFP into each. The majority of eGFP expressing NTS cells contained neuronal marker protein gene product 9.5 (PGP9.5)‐immunoreactivity (IR) but not glial marker glial fibrillary acidic protein (GFAP)‐IR. Similarly, the majority of eGFP expressing neurons in the PPG contained PGP9.5‐IR. We conclude that its neuronal selectivity and affinity make AAV2 a superior viral vector for transduction of NTS and PPG neurons.