Inayama Y, Tomiyama I, Kitamura H, Nakatani Y, Ito T, Nozawa A, Usuda Y, Kanisawa M
Department of Surgery, Yokohama City University School of Medicine, Japan.
Histochem Cell Biol. 1995 Sep;104(3):191-8. doi: 10.1007/BF01835152.
The purpose of this study was to investigate alkaline phosphatase (ALPase) reactivity in rabbit airway epithelial cells. Acetone-fixed, methyl benzoate and xylene-cleared (AMeX-treated) paraffin sections of trachea, bronchus, and lung tissue were stained by an azo dye coupling method for ALPase and examined by light microscopy. Electron histochemical staining was also performed in order to study the sensitivity and specificity of reactivity in each cell type. ALPase reactivity at the light microscopic level was observed exclusively in tracheo-bronchial basal cells, and not in bronchiolar basal cells. By electron microscopy, ALPase reactivity was noted in 97.9% of basal cells in the trachea, 97.0% of basal cells in the bronchus, and 94.5% of basal cells and 15.4% of Clara cells in the bronchiole. This was also true for dispersed tracheal epithelial cells. Reactivity was rarely observed in ciliated cells, non-goblet-type secretory cells, and undetermined cells. The reactivity was heat-labile, levamisole-sensitive, and of a non-specific type. These findings indicate that basal cells of rabbit trachea and bonchus have fairly high specificity for ALPase of a non-specific isozyme (92.2% and 95.6%, respectively). Therefore, ALPase is considered to be a useful marker for these cells.
本研究的目的是调查兔气道上皮细胞中的碱性磷酸酶(ALPase)活性。采用偶氮染料偶联法对气管、支气管和肺组织经丙酮固定、苯甲酸甲酯和二甲苯透明(AMeX处理)的石蜡切片进行ALPase染色,并通过光学显微镜检查。还进行了电子组织化学染色,以研究每种细胞类型中反应性的敏感性和特异性。在光学显微镜水平上,仅在气管-支气管基底细胞中观察到ALPase活性,而在细支气管基底细胞中未观察到。通过电子显微镜观察,气管中97.9%的基底细胞、支气管中97.0%的基底细胞、细支气管中94.5%的基底细胞和15.4%的克拉拉细胞中观察到ALPase活性。分散的气管上皮细胞也是如此。在纤毛细胞、非杯状分泌细胞和未确定细胞中很少观察到反应性。该反应性对热不稳定,对左旋咪唑敏感,且为非特异性类型。这些发现表明,兔气管和支气管的基底细胞对非特异性同工酶的ALPase具有相当高的特异性(分别为92.2%和95.6%)。因此,ALPase被认为是这些细胞的一种有用标志物。