Morita T, Usuda N, Hanai T, Nagata T
Department of Anatomy and Cell Biology, Shinshu University School of Medicine, Matsumoto, Japan.
Histochemistry. 1994 Jan;101(1):13-20. doi: 10.1007/BF00315826.
We report a change in the proliferative activity of mouse colonic epithelium due to development and aging. In order to measure the proliferative activity, colonic epithelium was immunostained for cyclin proliferating cell nuclear antigen (PCNA/cyclin), which appears from the Gl to the S phase of the cell cycle, and compared with labeling obtained by [3H]-thymidine radioautography. Litter mice of six age groups from the fetal period (embryonic day 19), newborn period (postnatal day 1), suckling period (postnatal day 5), weaning period (postnatal dy 21), adult period (2 month old) to the senescent period (11 month old) were examined by immunohistochemistry. The descending colons were fixed in methacarn (method-Carnoy) and embedded in paraffin. Sections were stained for PCNA/cyclin activity using 19A2 monoclonal antibody and the avidin-biotin peroxidase complex (ABC) technique. For radioautography, litter mice of nine age groups using in vivo intraperitoneal administration of [3H]-thymidine. The labeling indices of colonic epithelial cells in the proliferative zone were then analyzed and compared between the two investigative methods. Our results show that the proliferative activity of mice colon was high in the fetal and newborn periods and almost constant from the suckling period to senescence, as demonstrated by both PCNA/cyclin immunohistochemistry and [3H]-thymidine radioautography. The labeling index seen by PCNA/cyclin immunohistochemistry was, however, higher than that seen by [3H]-thymidine radioautography.
我们报告了小鼠结肠上皮细胞增殖活性随发育和衰老而发生的变化。为了测量增殖活性,对结肠上皮细胞进行免疫染色,检测细胞周期蛋白增殖细胞核抗原(PCNA/细胞周期蛋白),该抗原在细胞周期的G1期到S期出现,并与通过[3H] - 胸腺嘧啶核苷放射自显影获得的标记进行比较。通过免疫组织化学检查了六个年龄组的同窝小鼠,从胎儿期(胚胎第19天)、新生期(出生后第1天)、哺乳期(出生后第5天)、断奶期(出生后第21天)、成年期(2个月大)到衰老期(11个月大)。降结肠用甲醇 - 冰醋酸固定液(卡诺伊固定液)固定并石蜡包埋。切片使用19A2单克隆抗体和抗生物素蛋白 - 生物素过氧化物酶复合物(ABC)技术进行PCNA/细胞周期蛋白活性染色。对于放射自显影,九个年龄组的同窝小鼠采用体内腹腔注射[3H] - 胸腺嘧啶核苷。然后分析并比较两种研究方法中增殖区结肠上皮细胞的标记指数。我们的结果表明,通过PCNA/细胞周期蛋白免疫组织化学和[3H] - 胸腺嘧啶核苷放射自显影均显示,小鼠结肠的增殖活性在胎儿期和新生期较高,从哺乳期到衰老期几乎保持恒定。然而,PCNA/细胞周期蛋白免疫组织化学所见的标记指数高于[3H] - 胸腺嘧啶核苷放射自显影所见的标记指数。