Yamada K, Moriguchi A, Morishita R, Aoki M, Nakamura Y, Mikami H, Oshima T, Ninomiya M, Kaneda Y, Higaki J, Ogihara T
Department of Geriatric Medicine, Osaka University Medical School, Suita, Japan.
Am J Physiol. 1996 Nov;271(5 Pt 2):R1212-20. doi: 10.1152/ajpregu.1996.271.5.R1212.
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.
我们在体内和体外研究了用日本血凝病毒（HVJ）-脂质体载体递送后，寡脱氧核苷酸（ODN）在中枢神经系统（CNS）中的效率和细胞内命运。在大鼠小脑的原代培养颗粒细胞中，体外应用与HVJ-脂质体复合的异硫氰酸荧光素（FITC）标记的ODN导致强烈荧光定位于细胞核中，持续≥2周，而单独使用ODN时荧光仅持续3天。通过不同方法尝试进行体内ODN转移：注入下丘脑室旁核和侧脑室。通过HVJ-脂质体方法将FITC标记的ODN注入下丘脑，比单独注射ODN产生更高的浓度和更持久的荧光。用HVJ-脂质体将ODN注入侧脑室，在脑室周围层产生更明显和持久的荧光，主要在细胞核中。此外，用HVJ-脂质体方法荧光细胞的分布更广泛。这些结果表明，HVJ-脂质体方法延长了ODN的半衰期并将它们集中在细胞核中。因此，它是一种有效的ODN转移方法，并有望作为CNS中的基因递送方法。
