Differential adenoassociated virus vector-driven expression of a neuropeptide Y gene in primary rat brain astroglial cultures after transfection with Sendai virosomes versus Lipofectin.
The ability of Sendai virosomes or Lipofectin to introduce an AAV vector into primary rat brain astroglial cultures was characterized. The pJDT95npy vector was constructed by inserting rat NPY cDNA downstream from the indigenous AAV p5, p19 and p40 promoters in pJDT95. Lipofectin-mediated transfection with pJDT95npy (10 micrograms) resulted in pronounced expression of several NPY mRNA species: p5-driven (3.3 kb), p19-driven (2.7 kb) and p40-driven (0.6, 0.8, 1.1, and 1.8 kb). Exposure to virosomally encapsulated pJDT95npy (50 or 100 ng) resulted in transient expression of some p40-driven mRNA species (0.8 and 1.8 kb). Neither method produced astroglia cells which synthesized mature NPY immunoreactivity. This demonstrates that an AAV-derived vector can drive gene expression in astroglia, that Sendai virosomes can infuse vectors into astroglia, but that the amount of DNA infused in this manner may limit long term expression.
de Fiebre
C M
CM
Department of Pharmacology and Therapeutics, University of Florida College of Medicine, J. Hillis Miller Health Center, Gainesville 32610-0267.
Wu
P
P
Notabartolo
D
D
Millard
W J
WJ
Meyer
E M
EM
D000818:Animals; D001253:Astrocytes / Q000166:cytology / Q000378:metabolism*; D015152:Blotting, Northern; D001921:Brain / Q000378:metabolism*; D002478:Cells, Cultured; D000229:Dependovirus / Q000235:genetics*; D015870:Gene Expression; D005822:Genetic Vectors; D007700:Kinetics; D009478:Neuropeptide Y / Q000096:biosynthesis*; D010222:Parainfluenza Virus 1, Human / Q000235:genetics*; D010714:Phosphatidylethanolamines; D012333:RNA, Messenger / Q000032:analysis / Q000096:biosynthesis; D051381:Rats; D013997:Time Factors; D014162:Transfection
D009478:Neuropeptide Y; D010714:Phosphatidylethanolamines; D012333:RNA, Messenger; C020888:1,2-dielaidoylphosphatidylethanolamine