Sick T J, Perez-Pinzon M A
Department of Neurology, University of Miami School of Medicine, South Campus, Building B, 12500 Southwest 152 Street, Miami, Florida 33177, USA.
Methods. 1999 Jun;18(2):104-8. doi: 10.1006/meth.1999.0763.
Brain slice preparations have become useful tools for studying multiple facets of normal brain function and for investigations of brain pathophysiology. Recently, a variety of neurological disorders have been linked to dysfunction of brain mitochondria. In this report we discuss optical methods for probing mitochondrial function in brain slices. Absorption spectrophotometric and spectrofluorometric techniques are described for measuring changes in the redox activity of mitochondrial cytochromes and the primary respiratory chain substrate nicotinamide adenine dinucleotide (NADH), respectively. A spectrofluorometric method is described also for measuring changes in mitochondrial membrane potential using the potential-sensitive fluorescent indicator JC-1. These methods used together have proven to be useful for studying dysfunction of mitochondria following in vitro ischemia in hippocampal slices, and might also be valuable for investigations of mitochondrial involvement in other neurological disorders.
脑片制备已成为研究正常脑功能多个方面以及脑病理生理学的有用工具。最近,多种神经系统疾病已被发现与脑线粒体功能障碍有关。在本报告中,我们讨论了用于探测脑片中线粒体功能的光学方法。分别描述了吸收分光光度法和荧光分光光度法,用于测量线粒体细胞色素的氧化还原活性变化和主要呼吸链底物烟酰胺腺嘌呤二核苷酸(NADH)的变化。还描述了一种荧光分光光度法,使用电位敏感荧光指示剂JC-1测量线粒体膜电位的变化。这些方法一起使用已被证明可用于研究海马体切片体外缺血后的线粒体功能障碍,并且对于研究线粒体在其他神经系统疾病中的作用可能也具有重要价值。