Sendai virosomal infusion of an adeno-associated virus-derived construct containing neuropeptide Y into primary rat brain cultures.
A novel neuronal gene-delivery system was investigated in primary neuron-enriched cultures with respect to driving the expression of neuropeptide Y (NPY). This delivery system consists of an adeno-associated virus-derived (AAV) plasmid, pJDT95npy, encapsulated in reconstituted Sendai virosomes. pJDT95npy contains full length rat NPY cDNA inserted downstream from the P40 promoter in a cap-gene deleted AAV-derived construct. The rep-sequences under control of the P5 and P19 promoters are intact. Virosomally encapsulated pJDT95npy drove the expression of NPY mRNAs, predominantly by P40. Total cellular NPY immunoreactivity and release in the presence of depolarization increased following pJDT95npy-transfection. Neither empty virosomes nor virosomes containing pJDT95 affected NPY mRNA expression or immunoreactivity. This study demonstrates that an AAV-derived plasmid can drive exogenous gene expression in intact neurons after infusion by Sendai virosomes.
Wu
P
P
Department of Pharmacology and Therapeutics, University of Florida College of Medicine, J. Hillis Miller Health Center, Gainesville 32610, USA.
de Fiebre
C M
CM
Millard
W J
WJ
Elmstrom
K
K
Gao
Y
Y
Meyer
E M
EM
D000256:Adenoviridae / Q000235:genetics*; D000818:Animals; D001923:Brain Chemistry / Q000502:physiology*; D002478:Cells, Cultured; D003470:Culture Media; D005822:Genetic Vectors; D009478:Neuropeptide Y / Q000235:genetics / Q000378:metabolism*; D010222:Parainfluenza Virus 1, Human / Q000235:genetics*; D010957:Plasmids; D011401:Promoter Regions, Genetic; D012313:RNA / Q000096:biosynthesis; D051381:Rats; D014162:Transfection
D003470:Culture Media; D009478:Neuropeptide Y; D012313:RNA