Meijer A E
Histochem J. 1985 Nov;17(11):1271-5. doi: 10.1007/BF01002508.
In this communication an enzyme histochemical multistep technique for the demonstration of class 1 fructose-1,6-diphosphate aldolase in heart and skeletal muscle sections is described. With this technique a semipermeable membrane is interposed between the incubating solution and the tissue sections preventing diffusion of the enzyme into the medium during incubation. In the histochemical system the enzyme cleaves the substrate D-fructose-1,6-diphosphate to dihydroxyacetone phosphate and D-glyceraldehyde-3-phosphate. The dihydroxyacetone phosphate is reversibly converted into D-glyceraldehyde-3-phosphate by exogenous and endogenous triose phosphate isomerase. Next the D-glyceraldehyde-3-phosphate is oxidized by exogenous and endogenous glyceraldehyde-3-phosphate dehydrogenase and the electrons are transported concomitantly via NAD+, phenazine methosulphate and menadione to nitro-BT. Sodium azide and amytal are incorporated to block electron transfer to the cytochromes.
本文描述了一种酶组织化学多步技术，用于在心脏和骨骼肌切片中显示1类果糖-1,6-二磷酸醛缩酶。采用该技术时，在孵育溶液和组织切片之间插入半透膜，防止孵育过程中酶扩散到培养基中。在组织化学系统中，该酶将底物D-果糖-1,6-二磷酸裂解为磷酸二羟丙酮和D-甘油醛-3-磷酸。磷酸二羟丙酮通过外源性和内源性磷酸丙糖异构酶可逆地转化为D-甘油醛-3-磷酸。接下来，D-甘油醛-3-磷酸被外源性和内源性甘油醛-3-磷酸脱氢酶氧化，电子通过NAD +、吩嗪硫酸甲酯和甲萘醌同时传递给硝基蓝四唑。加入叠氮化钠和戊巴比妥以阻断电子向细胞色素的传递。
