豚鼠耳蜗毛细胞线粒体中乳酸脱氢酶、琥珀酸脱氢酶、二氢烟酰胺腺嘌呤二核苷酸黄递酶和细胞色素氧化酶活性的超微结构细胞化学

The ultrastructural cytochemistry of lactic dehydrogenase, succinic dehydrogenase, dihydro-nicotinamide adenine dinucleotide diaphorase and cytochrome oxidase activities in hair cell mitochondria of the guinea pig cochlea.

作者信息

Spector G J

出版信息

J Histochem Cytochem. 1975 Mar;23(3):216-34. doi: 10.1177/23.3.236341.

DOI:10.1177/23.3.236341

Abstract

The use of cinnamyl nitroblue tetrazolium chloride (DS-NBT) in dehydrogenase experiments (lactic dehydrogenase, succinic dehydrogenase, nicotinamide adenine dinucleotide diaphorase) and 3,3'-diaminobenzidine tetrahydrochloride (DAB) in cytochrome oxidase experiments indicated that mitochondrial oxidoreduction reactions from nicotinamide adenine dinucleotide to cytochrome oxidase are located on the inner mitochondrial membrane in the outer compartment and the intracristate spaces. These reactions behave according to the chemiosmotic hypothesis. The cochlear hair cell mitochondria are cytochemically indistinguishable from free liver mitochondria. The heterogeneous mitochondrial staining pattern is related to the osmolarity of the incubation media, solubility of the enzymes and pH of the medium, but not to the fixation method.

摘要

在脱氢酶实验（乳酸脱氢酶、琥珀酸脱氢酶、烟酰胺腺嘌呤二核苷酸黄递酶）中使用肉桂基硝基蓝四氮唑氯化物（DS-NBT）以及在细胞色素氧化酶实验中使用3,3'-二氨基联苯胺四盐酸盐（DAB）表明，从烟酰胺腺嘌呤二核苷酸到细胞色素氧化酶的线粒体氧化还原反应位于线粒体外室和嵴内空间的线粒体内膜上。这些反应符合化学渗透假说。耳蜗毛细胞线粒体在细胞化学上与游离肝线粒体无法区分。线粒体染色模式的异质性与孵育介质的渗透压、酶的溶解度和介质的pH值有关，而与固定方法无关。