I will construct two AAV vectors to express ARL13B:mcherry and dnKif3a-ires-GFP. The constructs will contain fluorescent proteins to permit their visualization through microscopy. I will clone these constructs into a commonly used AAV vector in the McIntyre lab, verify their nucleotide sequence, and test the constructs in a cell culture system. Upon successful completion, I will use the Ocular Gene Therapy Core to generate the AAV. To test expression in vivo, AAV will be injected into the brains of adult rats, targeting the nucleus accumbens, olfactory bulb, and ventral tegmental area (VTA). One hemisphere of each brain will receive an injection of a virus containing either ARL13B:mcherry or dnKif3a, and the other hemisphere will receive a control virus. After four weeks, I will perfuse the rats and dissect their brains to perform cryosectioning. I will use immunohistochemistry to detect cilia markers such as ARL13B, AC3, and SSTR3 combined with cell type markers. I will collect images for data analysis using confocal microscopy. Ultimately, this project will determine if adult neuronal cilia can be manipulated in vivo through viral mediated approaches. This verification step is an important precursor before behavioral experiments can be performed in rats injected with these viruses.