Ogawa S, Brown H E, Okano H J, Pfaff D W
Laboratory of Neurobiology and Behavior, Rockefeller University, New York, NY 10021, USA.
Regul Pept. 1995 Oct 20;59(2):143-9. doi: 10.1016/0167-0115(95)00096-t.
Intracerebral diffusion, cellular uptake and intracellular localization of oligodeoxynucleotides (ODN) after their microinjection in mouse brains were examined. Using either tetramethylrhodamine-5-(and -6)-isothiocyanate (TRITC)- or gamma-33P ATP-labeled ODNs, it was found that both phosphodiester ODNs (D-ODN) and phosphorothioate ODNs (S-ODN) quickly diffused (up to about 500 microns) and were taken up by many cells around the injection site as early as 15 min after administration. Fluorescence labeling intensity and silver grain accumulation of D-ODNs were greatly reduced by 4 h after injection, whereas those of S-ODNs were stable beyond at least 8 h after injection. Most of labeled ODNs were found in neuronal cells as identified by immunocytochemistry for neurofilament, NF 200, and to a much lesser extent in astrocytic cells as identified by immunocytochemistry for glial fibrillary acidic protein.
研究了寡脱氧核苷酸（ODN）微量注射到小鼠脑内后的脑内扩散、细胞摄取及细胞内定位。使用异硫氰酸四甲基罗丹明-5-（及-6-）（TRITC）或γ-33P ATP标记的ODN，发现磷酸二酯ODN（D-ODN）和硫代磷酸酯ODN（S-ODN）均迅速扩散（可达约500微米），且在给药后15分钟时就被注射部位周围的许多细胞摄取。注射后4小时，D-ODN的荧光标记强度和银颗粒积累显著降低，而S-ODN的荧光标记强度和银颗粒积累在注射后至少8小时内保持稳定。通过针对神经丝、NF 200的免疫细胞化学鉴定发现，大多数标记的ODN存在于神经元细胞中，而通过针对胶质纤维酸性蛋白的免疫细胞化学鉴定发现，在星形胶质细胞中的含量要少得多。
