Hara H, Moriki T, Hiroi M, Yamane T
Acta Pathol Jpn. 1984 Sep;34(5):1049-57. doi: 10.1111/j.1440-1827.1984.tb07635.x.
Combination of autoradiography and ultracytochemical acridine orange method in rat astrocytoma C6 cells is described. Electron microscopic examination revealed that AO binds to DNA exclusively within the euchromatin portion of the cell nucleus. X-ray microanalysis showed that AO reaction products contain lead, uranium, and osmium. These heavy metals seem to contribute to electron density of the AO reaction products. The percentage of AO positive cells was 51.8 +/- 1.9%, whereas average labeling index of C6 cells incubated with 3H-thymidine for 30 minutes was 25.6 +/- 1.7%. About 82% of 3H-thymidine-labeled cell nuclei were AO positive and the remaining 18% were free of AO reaction products. On the other hand, about 27% of AO positive cells were not labeled with 3H-thymidine. However, most of C6 cell nuclei with active 3H-uridine incorporation showed AO chromatin reaction products. The results suggest that visualization of AO chromatin reaction products might be related to RNA synthesis rather than DNA synthesis in the cell nuclei.
本文描述了放射自显影与吖啶橙超微细胞化学方法在大鼠星形细胞瘤C6细胞中的联合应用。电子显微镜检查显示，吖啶橙仅与细胞核常染色质部分的DNA结合。X射线微分析表明，吖啶橙反应产物含有铅、铀和锇。这些重金属似乎有助于吖啶橙反应产物的电子密度。吖啶橙阳性细胞的百分比为51.8±1.9%，而用3H-胸腺嘧啶核苷孵育30分钟的C6细胞的平均标记指数为25.6±1.7%。约82%的3H-胸腺嘧啶核苷标记的细胞核为吖啶橙阳性，其余18%没有吖啶橙反应产物。另一方面，约27%的吖啶橙阳性细胞未被3H-胸腺嘧啶核苷标记。然而，大多数有活跃3H-尿苷掺入的C6细胞核显示有吖啶橙染色质反应产物。结果表明，吖啶橙染色质反应产物的可视化可能与细胞核中的RNA合成而非DNA合成有关。
