Histochemistry of sarcoplasmic reticulum Ca-ATPase using dysprosium as capturing reagent.

The Histochemical Journal Pub Date : 1995-09-01
W J van der Laarse, P van Noort, W S Simonides, P C Diegenbach, M B Lee-de Groot, C van Hardeveld
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Abstract

This report describes the development of a histochemical method for the demonstration of sarcoplasmic reticulum Ca-ATPase activity in cross-sections of skeletal muscle. The demonstration of sarcoplasmic reticulum Ca-ATPase activity is complicated by the fact that capturing reagents for phosphate inhibit the enzyme. We present a minimal model for heavy-metal-phosphate precipitation reactions which gives a theoretical description of the effect of enzyme inhibition on the rate of phosphate precipitation in the section. The model indicates that the choice of capturing reagent is crucial: whether or not ATPase activity can be demonstrated depends mainly on the inhibition constant and the solubility product of the phosphate salt of the capturing reagent (but also on a fairly large number of other factors). All lanthanides tested can be used to demonstrate sarcoplasmic reticulum Ca-ATPase activity, but dysprosium results in the highest staining intensity. This suggests that dysprosium inhibits sarcoplasmic reticulum Ca-ATPase to a lesser degree than the other lanthanides and/or the solubility product of its phosphate salt is smaller. As an example, the method is used to investigate the effect of thyroid hormone on sarcoplasmic reticulum Ca-ATPase activity in individual fibres of the rat soleus muscle.

肌浆网ca - atp酶的组织化学。
本报告描述了骨骼肌横截面肌浆网ca - atp酶活性的组织化学方法的发展。肌浆网ca - atp酶活性的证明由于磷酸捕获试剂抑制酶而变得复杂。我们提出了重金属-磷酸盐沉淀反应的最小模型,该模型理论上描述了酶抑制对该部分磷酸盐沉淀速率的影响。该模型表明,捕获试剂的选择是至关重要的:能否证明atp酶活性主要取决于捕获试剂的抑制常数和磷酸盐的溶解度积(但也取决于相当多的其他因素)。所有的镧系元素测试都可以用来证明肌浆网ca - atp酶活性,但镝的染色强度最高。这表明镝对肌浆网ca - atp酶的抑制程度低于其他镧系元素和/或其磷酸盐的溶解度产物较小。以该方法为例,研究了甲状腺激素对大鼠比目鱼肌肌浆网ca - atp酶活性的影响。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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