Mancini M A, Unakar N J, Giblin F J, Reddan J R
Department of Biological Sciences, Oakland University, Rochester, Mich.
Ophthalmic Res. 1989;21(5):369-73. doi: 10.1159/000266901.
The diaminobenzidine (DAB) technique was used to investigate the localization of the peroxidatic activity of catalase in cultured lens epithelial cells at the ultrastructural level. Cultured rabbit, bovine and mouse lens epithelial cells incubated in an alkaline DAB reaction mixture contained catalase-positive microperoxisomes which were randomly distributed throughout the cytoplasm. The reaction product of catalase was abolished when cells were treated with 3-amino-1-H-1,2,4-triazole, a specific inhibitor of catalase, and was not detected when DAB or H2O2 was omitted from the incubation medium, or when the pH of the reaction mixture was lowered to 7.0. The fine structure of lens epithelial cells that were constantly exposed to 0.025 or 0.05 mM hydrogen peroxide was also determined. Lens epithelial cells that were constantly exposed to 0.05 mM H2O2 for 1-3 h exhibited damaged mitochondria, a phenomenon that did not occur when the cells were exposed to 0.025 mM peroxide.
采用二氨基联苯胺(DAB)技术在超微结构水平上研究过氧化氢酶的过氧化物酶活性在培养的晶状体上皮细胞中的定位。在碱性DAB反应混合物中孵育的培养兔、牛和小鼠晶状体上皮细胞含有过氧化氢酶阳性的微过氧化物酶体,它们随机分布于整个细胞质中。当用过氧化氢酶的特异性抑制剂3-氨基-1-H-1,2,4-三唑处理细胞时,过氧化氢酶的反应产物消失,当孵育培养基中省略DAB或H2O2,或当反应混合物的pH值降至7.0时,未检测到反应产物。还测定了持续暴露于0.025或0.05 mM过氧化氢的晶状体上皮细胞的精细结构。持续暴露于0.05 mM H2O2 1-3小时的晶状体上皮细胞表现出线粒体受损,而当细胞暴露于0.025 mM过氧化物时未出现这种现象。