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锰超氧化物歧化酶在大鼠耳蜗中的免疫组织化学定位

Immunohistochemical localization of manganese superoxide dismutase in the rat cochlea.

作者信息

Lai M T, Ohmichi T, Egusa K, Okada S, Masuda Y

机构信息

Department of Otolaryngology, Okayama University Medical School, Japan.

出版信息

Eur Arch Otorhinolaryngol. 1996;253(4-5):273-7. doi: 10.1007/BF00171142.

Abstract

There has been recent increasing interest in the involvement of superoxide radicals (O2-) and their scavenging enzymes, the superoxide dismutases, in the patho-physiology of certain diseases. Since mitochondria are significant intracellular sources of O2- and important targets of oxidant injury, determining the intracochlear localization of mitochondrial O2- scavenging enzyme may provide important insight into the pathogenesis of injury due to cochlear oxidants. In order to locate the mitochondrial O2- scavenging enzyme, manganese superoxide dismutase (MnSOD), the authors used a modified immunoglobulin peroxidase bridge sequence method to detect MnSOD in paraffin-embedded, formalin-fixed rat cochleas. Site-specific immunolocalization of MnSOD could be demonstrated in the cochlear labyrinth, suggesting that the generation of intracochlear O2- was possibly implicated in the metabolically active sites and sites rich in vascularity. This study also provided a useful probe for detecting MnSOD immunohistochemically from ethylenediamine tetra-acetic acid-treated materials without requiring an antigen retrieval procedure.

摘要

最近,超氧阴离子自由基(O2-)及其清除酶——超氧化物歧化酶在某些疾病病理生理学中的作用越来越受到关注。由于线粒体是细胞内O2-的重要来源以及氧化损伤的重要靶点,确定耳蜗内线粒体O2-清除酶的定位可能为深入了解耳蜗氧化剂所致损伤的发病机制提供重要线索。为了定位线粒体O2-清除酶——锰超氧化物歧化酶(MnSOD),作者采用改良的免疫球蛋白过氧化物酶桥联序列法,在福尔马林固定、石蜡包埋的大鼠耳蜗中检测MnSOD。MnSOD的位点特异性免疫定位可在耳蜗迷路中得到证实,这表明耳蜗内O2-的产生可能与代谢活跃部位和血管丰富部位有关。该研究还提供了一种有用的探针,可从乙二胺四乙酸处理的材料中免疫组织化学检测MnSOD,而无需抗原修复程序。

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