一种用于碳酸酐酶组织化学定位的快速筛选方法。应用于肾脏、骨骼肌和血小板。

A fast screening method for histochemical localization of carbonic anhydrase. Application to kidney, skeletal muscle, and thrombocytes.

作者信息

Dermietzel R, Leibstein A, Siffert W, Zamboglou N, Gros G

出版信息

J Histochem Cytochem. 1985 Feb;33(2):93-8. doi: 10.1177/33.2.3918097.

DOI:10.1177/33.2.3918097

Abstract

A simple method for histochemical localization of carbonic anhydrase using 5-dimethyl-amino-naphthalene-1-sulfonamide (DNSA) is described. Cryosections of tissues, or cell smears, are incubated in 3 to 10 X 10(-5) M DNSA and viewed in a fluorescence microscope. Upon excitation with ultraviolet light, sites of carbonic anhydrase localization can be identified by an intense blue fluorescence, which is due to the emission of blue light (lambda max = 470 nm) by carbonic anhydrase-DNSA complexes. This fluorescence can be largely suppressed by simultaneous incubation with 1 X 10(-4) to 2 X 10(-3) M concentrations of nonfluorescent carbonic anhydrase inhibitors, displacing DNSA from its binding site on the enzyme. Application of the method to kidney, skeletal muscle, and thrombocytes yields patterns of carbonic anhydrase localization that are in good agreement with results that have been obtained with a variety of other techniques.

摘要

本文描述了一种使用5-二甲基氨基萘-1-磺酰胺（DNSA）对碳酸酐酶进行组织化学定位的简单方法。将组织冷冻切片或细胞涂片在3至10×10⁻⁵ M的DNSA中孵育，然后在荧光显微镜下观察。用紫外光激发后，碳酸酐酶定位的部位可通过强烈的蓝色荧光识别，这是由于碳酸酐酶-DNSA复合物发出蓝光（最大波长=470 nm）所致。同时与1×10⁻⁴至2×10⁻³ M浓度的非荧光碳酸酐酶抑制剂孵育，可使DNSA从其在酶上的结合位点被置换，从而大大抑制这种荧光。将该方法应用于肾脏、骨骼肌和血小板，得到的碳酸酐酶定位模式与使用多种其他技术获得的结果高度一致。