Balaban R S, Dennis V W, Mandel L J
Am J Physiol. 1981 Apr;240(4):F337-42. doi: 10.1152/ajprenal.1981.240.4.F337.
We report the successful application of microfluorometric techniques to monitor the redox state of mitochondrial nicotinamide adenine dinucleotide (NAD) in conjunction with measurements of transepithelial transport in intact isolated perfused tubules from the rabbit kidney. The determination of the redox state of NAD yields important information on the delivery of substrate to, and the energy demands on, the tubular mitochondria. These studies required only minor modifications of the basic perfusion apparatus. With this technique we detected net reductions of mitochondrial NAD upon the addition of metabolic substrates (succinate), the inhibition of active ion transport (ouabain), and the inhibition of mitochondrial electron flow (cyanide). Uncoupling of oxidative phosphorylation with 1799 caused an oxidation of NAD. These changes in NAD redox state are predictable from previous studies in isolated mitochondria, intact tissues, and from experiments on cortical tubule suspensions from the rabbit cortex.
我们报告了微荧光技术的成功应用，该技术用于监测线粒体烟酰胺腺嘌呤二核苷酸（NAD）的氧化还原状态，并结合对兔肾完整分离灌注小管跨上皮转运的测量。NAD氧化还原状态的测定可提供关于底物向肾小管线粒体的输送以及线粒体能量需求的重要信息。这些研究仅需对基本灌注装置进行微小改动。通过该技术，我们检测到添加代谢底物（琥珀酸）、抑制主动离子转运（哇巴因）以及抑制线粒体电子流（氰化物）后线粒体NAD的净还原。用1799使氧化磷酸化解偶联导致NAD氧化。NAD氧化还原状态的这些变化可从先前对分离线粒体、完整组织的研究以及对兔皮质皮质小管悬浮液的实验中预测出来。
