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硬骨鱼肾细胞微体中过氧化氢酶和D-氨基酸氧化酶的细胞化学显示

The cytochemical demonstration of catalase and D-amino acid oxidase in the microbodies of teleost kidney cells.

作者信息

Veenhuis M, Bonga S D

出版信息

Histochem J. 1977 Mar;9(2):171-81. doi: 10.1007/BF01003629.

Abstract

The distribution of catalase and D-amino acid oxidase, marker enzymes for peroxisomes, was determined cytochemically in the kidney tubules of an euryhaline teleost, the three-spined stickleback. Catalase activity was localized with the diaminobenzidine technique. The presence of D-amino acid oxidase was determined using H2O2 generated by the enzyme, D-alanine as a substrate, and cerous ions for the formation of an electron-dense precipitate. Both enzymes appeared to be located in microbodies. The combined presence of these enzymes characterizes the microbodies as peroxisomes. Biochemically and cytochemically, no urate oxidase or glycolate-oxidizing L-alpha-hydroxy acid oxidase could be demonstrated. Stereological analysis of the epithelia lining the renal tubules showed that the fractional volume of the microbodies is 5 to 10 times higher in the cells of the second proximal tubules than in the other nephronic segments or the ureter. The fractional volume of the microbodies was similar in kidneys of freshwater and seawater fishes.

摘要

利用细胞化学方法,测定了广盐性硬骨鱼三刺鱼肾小管中过氧化物酶体的标记酶——过氧化氢酶和D-氨基酸氧化酶的分布情况。采用二氨基联苯胺技术定位过氧化氢酶活性。利用该酶产生的H2O2、以D-丙氨酸作为底物以及铈离子形成电子致密沉淀物的方法,来确定D-氨基酸氧化酶的存在。这两种酶似乎都位于微体中。这些酶的共同存在将微体表征为过氧化物酶体。在生物化学和细胞化学方面,均未证实存在尿酸氧化酶或氧化乙醇酸的L-α-羟基酸氧化酶。对肾小管内衬上皮的体视学分析表明,第二近端小管细胞中微体的分数体积比其他肾单位节段或输尿管中的高5至10倍。淡水鱼和海水鱼肾脏中微体的分数体积相似。

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