Efficient oligonucleotide delivery using the HVJ-liposome method in the central nervous system.
We examined the efficiency and intracellular fate of oligodeoxy-nucleotides (ODN) in the central nervous system (CNS) after delivery with a hemagglutinating virus of Japan (HVJ)-liposome vector in vivo and in vitro. In primary cultured granular cells of the rat cerebellum, application of fluorescein isothiocyanate (FITC)-labeled ODN complexed with HVJ-liposomes in vitro resulted in strong fluorescence localized in nuclei that persisted for > or = 2 wk, in contrast to 3 days with ODN alone. In vivo ODN transfer was attempted by different approaches: infusions into the paraventricular nuclei of the hypothalamus and the lateral cerebroventricle. Injection of FITC-labeled ODN into the hypothalamus by the HVJ-liposome method produced a higher concentration and more persistent fluorescence than did injection of ODN alone. Administration of ODN into the lateral cerebroventricle with HVJ-liposomes yielded more conspicuous and prolonged fluorescence in the periventricular layer, predominantly in cell nuclei. Furthermore, the distribution of fluorescent cells was broader with the HVJ-liposome method. These results indicate that the HVJ-liposome method prolongs the half-life of ODN and concentrates them in cell nuclei. Thus it is an efficient method for ODN transfer and holds promise as a gene delivery method in the CNS.
Yamada
K
K
Department of Geriatric Medicine, Osaka University Medical School, Suita, Japan.
Moriguchi
A
A
Morishita
R
R
Aoki
M
M
Nakamura
Y
Y
Mikami
H
H
Oshima
T
T
Ninomiya
M
M
Kaneda
Y
Y
Higaki
J
J
Ogihara
T
T
D000818:Animals; D001921:Brain / Q000166:cytology / Q000378:metabolism*; D002478:Cells, Cultured; D002531:Cerebellum / Q000166:cytology / Q000378:metabolism; D016650:Fluorescein-5-isothiocyanate; D005456:Fluorescent Dyes; D005822:Genetic Vectors; D008081:Liposomes; D008297:Male; D008722:Methods; D009841:Oligonucleotides / Q000008:administration & dosage* / Q000378:metabolism / Q000493:pharmacokinetics; D051381:Rats; D017207:Rats, Sprague-Dawley; D018108:Respirovirus
D005456:Fluorescent Dyes; D008081:Liposomes; D009841:Oligonucleotides; D016650:Fluorescein-5-isothiocyanate