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使用牛肝过氧化氢酶作为电子显微镜的蛋白质示踪剂。

The use of beef liver catalase as a protein tracer for electron microscopy.

作者信息

Venkatachalam M A, Fahimi H D

出版信息

J Cell Biol. 1969 Aug;42(2):480-9. doi: 10.1083/jcb.42.2.480.

Abstract

Beef liver catalase was injected intravenously into mice, and its distribution in the kidney, myocardium, and liver was studied with the electron microscope. A specific and relatively sensitive method was developed for its ultrastructural localization, based on the peroxidatic activity of catalase and employing a modified Graham and Karnovsky incubation medium. The main features of the medium were a higher concentration of diaminobenzidine, barium peroxide as the source of peroxide, and pH of 8.5. Ultrastructurally, the enzyme was seen to permeate the endothelial fenestrae and basement membranes of tubular and glomerular capillaries of the kidney. The urinary space and tubular lumina contained no reaction product. In the myocardial capillaries, the tracer filled the pinocytotic vesicles but did not diffuse across the intercellular clefts of the endothelium. In liver, uptake of catalase was seen both in hepatocytes and in Kupffer cells.

摘要

将牛肝过氧化氢酶静脉注射到小鼠体内,并用电子显微镜研究其在肾脏、心肌和肝脏中的分布。基于过氧化氢酶的过氧化物活性并采用改良的格雷厄姆和卡诺夫斯基孵育培养基,开发了一种用于其超微结构定位的特异性且相对灵敏的方法。该培养基的主要特点是二氨基联苯胺浓度更高、以过氧化钡作为过氧化物来源以及pH值为8.5。在超微结构上,可见该酶透过肾脏肾小管和肾小球毛细血管的内皮窗孔和基底膜。尿腔和肾小管管腔中没有反应产物。在心肌毛细血管中,示踪剂充满了胞饮小泡,但没有扩散穿过内皮细胞的细胞间裂隙。在肝脏中,肝细胞和库普弗细胞均可见过氧化氢酶的摄取。

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