Perotti M E, Gavazzi E, Trussardo L, Malgaretti N, Curti B
Histochem J. 1987 Mar;19(3):157-69. doi: 10.1007/BF01695140.
The intracellular localization of D-amino acid oxidase in rat kidney and liver has been investigated using the indirect immunogold postembedding technique. Different fixation and embedding conditions for optimal preservation of antigenicity and fine structure have been tested. Immunolabelling was possible only in tissues embedded in polar resins (glycol methacrylate and Lowicryl K4M). In kidney the enzyme was demonstrable only in the peroxisomes of the proximal tubule, where it was associated with the peroxisome core. The enzyme was present in all the peroxisomes of the proximal tubule and appeared to be codistributed with catalase. Control experiments and quantitative analysis confirmed the specificity of the D-amino acid oxidase immunolocalization. All the other cells in kidney failed to demonstrate any labelling. In liver, the immunolabelling was present in the matrix of the hepatocyte peroxisomes, whereas no traces of the enzyme were found in the nucleoid. The intensity of the immunolabelling in liver peroxisomes was lower than in kidney. No specific labelling was observed in cells other than hepatocytes.
利用间接免疫金包埋后技术研究了大鼠肾脏和肝脏中D-氨基酸氧化酶的细胞内定位。测试了不同的固定和包埋条件,以实现对抗原性和精细结构的最佳保存。仅在嵌入极性树脂(甲基丙烯酸乙二醇酯和低温包埋剂K4M)的组织中才能进行免疫标记。在肾脏中,该酶仅在近端小管的过氧化物酶体中可检测到,且与过氧化物酶体核心相关。该酶存在于近端小管的所有过氧化物酶体中,且似乎与过氧化氢酶共分布。对照实验和定量分析证实了D-氨基酸氧化酶免疫定位的特异性。肾脏中的所有其他细胞均未显示任何标记。在肝脏中,免疫标记存在于肝细胞过氧化物酶体的基质中,而在核样体中未发现该酶的踪迹。肝脏过氧化物酶体中的免疫标记强度低于肾脏。除肝细胞外,在其他细胞中未观察到特异性标记。