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) containing rat neuropeptide Y (NPY) cDNA inserted downstream of endogenous AAV promoters was used to investigate AAV-driven NPY expression in postmitotic neurons in vitro and in the brain. NPY mRNA was expressed in NT2/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 depolarization in the primary cultures. Injections of Sendai-virosome encapsulated pJDT95npy into neocortex increased NPY-like immunoreactivity in neurons but not glia indicating that the latter cell type did not have the translational, post-translational or storage capacity to accumulate the peptide. Injections into the rat hypothalamic para-ventricular nucleus increased body weight and food intake for 21 days, though NPY-like immunoreactivity remained elevated for at least 50 days. These studies demonstrate that AAV-derived constructs may be useful for delivering genes into post-mitotic neurons, and that Sendai virosomes are effective for delivering these constructs in vivo.
Wu
P
P
Department of Pharmacology and Therapeutics, University of Florida College of Medicine, USA.
de Fiebre
C M
CM
Millard
W J
WJ
King
M A
MA
Wang
S
S
Bryant
S O
SO
Gao
Y P
YP
Martin
E J
EJ
Meyer
E M
EM
D000818:Animals; D001921:Brain / Q000166:cytology*; D002478:Cells, Cultured; D000229:Dependovirus / Q000235:genetics*; D005260:Female; D008297:Male; D009474:Neurons / Q000166:cytology*; D009478:Neuropeptide Y / Q000235:genetics*; D010222:Parainfluenza Virus 1, Human / Q000235:genetics*; D011401:Promoter Regions, Genetic; D051381:Rats; D017207:Rats, Sprague-Dawley; D014162:Transfection
D009478:Neuropeptide Y