A QUANTITATIVE BIOCHEMICAL AND HISTOCHEMICAL STUDY OF THE LEAD METHOD FOR LOCALIZATION OF ADENOSINE TRIPHOSPHATE-HYDROLYZING ENZYMES
A QUANTITATIVE BIOCHEMICAL AND HISTOCHEMICAL STUDY OF THE LEAD METHOD FOR LOCALIZATION OF ADENOSINE TRIPHOSPHATE-HYDROLYZING ENZYMES
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三磷酸腺苷水解酶定位先导法的定量生化和组织化学研究
DOI：
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发表时间：
1969
影响因子：
3.2
通讯作者：
P. Jørgensen
中科院分区：
文献类型：
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作者：
N. O. Jacobsen;P. Jørgensen
目的：研究铅法定位于细胞膜的三磷酸腺苷酶（ATP酶）的性质及铅催化ATP酶水解对固定肾染色的意义。使用改良的Wachstein和Meisel培养基。本文比较了铅对大鼠肾微粒体沉积物切片的染色和铅对（Na ~+ + K ~+）-、Mg ~（++）-和Ca ~（++）-ATP酶活性的影响。在大鼠肾脏切片上检测这些观察结果的组织化学相关性。本地化的（Na+ + K+）-ATP酶是不可能的浓度低于失活水平（0.5毫米）的铅。Mg++-ATP酶和Ca++-ATP酶仅被3.6 mM铅部分抑制，并且在戊二醛固定的肾脏中定位于质膜。染色后的激活过程中的Ca++和Mg++的酶促水解的特点和激活ATP的非酶水解不同。固定组织的染色在2-3 mM铅时最大，并在较高浓度下降低。在Mg++缺乏的媒体膜染色是弱的或不存在的5.2-6.8 mM的铅，其中ATP的非酶水解率高。因此，在所使用的介质中，铅催化的ATP水解对固定肾的染色贡献很小。
The purpose was to study the nature of the adenosine triphosphatase (ATPase) localized to plasma membranes by the lead method and the significance of the lead-catalyzed hydrolysis of adenosine triphosphate (ATP) for the staining of fixed kidney. A modified Wachstein and Meisel's medium was used. The staining of sections of microsomal sediments from rat kidney was compared with the in vitro activity of (Na+ + K+)–, Mg++– and Ca++– ATPase in the presence of lead. The histochemical relevance of these observations was tested on sections of rat kidney. Localization of (Na+ + K+)–ATPase was not possible at concentrations of lead below the inactivating level (0.5 mM). Mg++–ATPase and Ca++– ATPase were only partially inhibited by 3.6 mM lead and were localized to plasma membranes in glutaraldehyde-fixed kidney. The staining followed a course of activation by Ca++ and Mg++ characteristic for enzymatic hydrolysis and different from the activation of nonenzymatic hydrolysis of ATP. Staining of fixed tissue was maximal at 2-3 mM lead and decreased at higher concentrations. In Mg++–deficient media membrane staining was weak or absent at 5.2-6.8 mM lead, where the rate of nonenzymatic hydrolysis of ATP was high. Thus, in the medium used the lead-catalyzed hydrolysis of ATP contributed little to the staining of fixed kidney.