Frederiks W M, Bosch K S, Ankum M, Wanders R J
Laboratory of Cell Biology and Histology, University of Amsterdam, The Netherlands.
J Inherit Metab Dis. 1993;16(6):921-8. doi: 10.1007/BF00711506.
The localization of the activity of the peroxisomal enzymes D-amino acid oxidase and hydroxyacid oxidase was studied at the light-microscopical level in livers and kidneys of control subjects and patients with Zellweger syndrome, an inherited disease characterized by a lack of intact peroxisomes. D-Amino acid oxidase and hydroxyacid oxidase activities were demonstrated in unfixed cryostat sections with the cerium-diaminobenzidine-cobalt-hydrogen peroxide procedure, in which cerium ions capture hydrogen peroxide, the product of both enzymes. In a second step reaction decomposition of cerium perhydroxide gives rise to a diaminobenzidine polymer complexed with cobalt ions. D-Amino acid oxidase and hydroxyacid oxidase activities were found in peroxisomes of liver parenchymal cells, and only D-amino acid oxidase in peroxisomes of proximal tubular cells of kidneys of control humans. The activities of these enzymes were not detectable in livers and kidneys of Zellweger patients. It is concluded that the cerium-diaminobenzidine-cobalt-hydrogen peroxide procedure enables the demonstration of peroxisomal enzyme activities in human tissues at the light-microscopical level and is an important tool in detecting patients with Zellweger syndrome.
在对照受试者以及患有齐-韦二氏综合征（一种以缺乏完整过氧化物酶体为特征的遗传性疾病）的患者的肝脏和肾脏中，利用光学显微镜研究了过氧化物酶体酶D-氨基酸氧化酶和羟酸氧化酶活性的定位。采用铈-二氨基联苯胺-钴-过氧化氢法在未固定的低温恒温器切片中显示D-氨基酸氧化酶和羟酸氧化酶活性，在该方法中铈离子捕获这两种酶的产物过氧化氢。在第二步反应中，过氢氧化铈的分解产生与钴离子络合的二氨基联苯胺聚合物。在对照人类肝脏实质细胞的过氧化物酶体中发现了D-氨基酸氧化酶和羟酸氧化酶活性，而在肾脏近端小管细胞的过氧化物酶体中仅发现了D-氨基酸氧化酶活性。在齐-韦二氏综合征患者的肝脏和肾脏中未检测到这些酶的活性。得出的结论是，铈-二氨基联苯胺-钴-过氧化氢法能够在光学显微镜水平上显示人体组织中的过氧化物酶体酶活性，并且是检测齐-韦二氏综合征患者的重要工具。
