AN AAV PROMOTER-DRIVEN NEUROPEPTIDE-Y GENE DELIVERY SYSTEM USING SENDAI VIROSOMES FOR NEURONS AND RAT-BRAIN

An adeno-associated virus (AAV)-derived construct (pJDT95npy) containi ng rat neuropeptide Y (NPY) cDNA inserted downstream of endogenous AAV promoters was used to investigate AAV-driven NPY expression in post-m itotic neurons in vitro and in the brain. NPY mRNA was expressed in NT 2/N and rat brain primary neuronal cultures after transfection. There was a corresponding increase in the number of neurons staining for NPY -like immunoreactivity and an increase in NPY release during depolariz ation in the primary cultures. Injections of Sendai-virosome encapsula ted pJDT95npy into neocortex increased NPY-like immunoreactivity in ne urons but not glia indicating that the latter cell type did not have t he translational, post-translational or storage capacity to accumulate the peptide. injections into the rat hypothalamic paraventricular nuc leus increased body weight and food intake for 21 days, though NPY-lik e immunoreactivity remained elevated for at least 50 days. These studi es demonstrate that AAV-derived constructs may be useful for deliverin g genes into post-mitotic neurons, and that Sendai virosomes are effec tive for delivering these constructs in vivo.