Yokota S, Himeno M, Kato K
Department of Anatomy, Yamanashi Medical School, Fukuoka, Japan.
Cell Struct Funct. 1989 Apr;14(2):163-71. doi: 10.1247/csf.14.163.
Localization of acid phosphatase (ACPase) in rat liver was investigated by immunocytochemical techniques. Rat liver was fixed by perfusion and cut into thick tissue slices, which were embedded in Epon or Lowicryl K4M. For light microscopy (LM), semithin Epon sections were stained for the enzyme ACPase by an indirect immunoenzyme technique. For electron microscopy (EM), ultra-thin Lowicryl K4M sections were stained by a protein A-gold technique. By means of LM, granular reaction deposits were observed in hepatocytes and sinus-lining cells. Stained granules were present in the juxtanuclear cytoplasm, but they did not correspond to a typical staining pattern for the Golgi complex. EM revealed that gold particles indicating ACPase antigens were present on lysosomes and on some vesicles locating in the trans Golgi region. Endosomelike vesicles were strongly positive for the labeling. Golgi cisterna were mostly negative, but weak signals were noted in dilated sacules. The plasma membranes on the sinusoidal and bile canalicular sides were labeled by a few gold particles. The results indicate that ACPase is present in endosomes and in a restricted area of plasma membrane, as well as in the lysosomal system.
采用免疫细胞化学技术研究了大鼠肝脏中酸性磷酸酶(ACPase)的定位。通过灌注法固定大鼠肝脏并切成厚组织切片,将其包埋于环氧树脂或Lowicryl K4M中。对于光学显微镜(LM),用间接免疫酶技术对环氧树脂半薄切片进行酸性磷酸酶染色。对于电子显微镜(EM),用蛋白A-金技术对Lowicryl K4M超薄切片进行染色。借助光学显微镜,在肝细胞和窦衬细胞中观察到颗粒状反应沉积物。染色颗粒存在于核周细胞质中,但它们与高尔基体复合体的典型染色模式不符。电子显微镜显示,指示酸性磷酸酶抗原的金颗粒存在于溶酶体和位于高尔基体反面膜囊区域的一些小泡上。内体样小泡标记呈强阳性。高尔基体扁平囊大多呈阴性,但在扩张的囊泡中观察到微弱信号。窦状隙侧和胆小管侧的质膜上有少量金颗粒标记。结果表明,酸性磷酸酶存在于内体、质膜的特定区域以及溶酶体系统中。