Ben-Shachar Dorit, Suss-Toby Edith, Robicsek Odile
Laboratory of Psychobiology, Department of Psychiatry, B. Rappaport Faculty of Medicine, Rambam Medical Center, Technion IIT, 9649, Haifa, 31096, Israel,
Methods Mol Biol. 2015;1265:425-39. doi: 10.1007/978-1-4939-2288-8_32.
Mitochondria, similar to living cells and organelles, have negative membrane potential and can therefore accumulate permeable lipophilic cations. Those cations which exhibit fluorescence activity after accumulation into energized systems are widely used to decipher changes in membrane potential by imaging techniques. Here we describe the use of the lipophilic cation 5,5',6,6'tetrachloro-1,1',3,3'-tetraethylbenzimidazol-carbocyanine iodide (JC-1), which alters reversibly its color from green (J-monomer, at its low concentration in the cytosol) to red (J-aggregates, at its high concentration in active mitochondria) with increasing mitochondrial membrane potential (Δψm). We show that in addition to changes in Δψm, this specific dye can be used to follow alterations in mitochondrial distribution and mitochondrial network connectivity. We suggest that JC-1 is a preferable probe to compare between treatment groups, as the ratio of green to red fluorescence intensities is used for analysis. This ratio depends only on the mitochondrial membrane potential and not on other mitochondrial dependent or independent factors. We demonstrate various applications of JC-1 staining to study mitochondrial abnormalities in different cell types derived from schizophrenia patients and healthy subjects.
线粒体与活细胞及细胞器类似，具有负膜电位，因此能够积累可渗透的亲脂性阳离子。那些在积累到活跃系统后表现出荧光活性的阳离子被广泛用于通过成像技术来解读膜电位的变化。在此，我们描述了亲脂性阳离子5,5',6,6'-四氯-1,1',3,3'-四乙基苯并咪唑羰花青碘化物（JC-1）的用途，随着线粒体膜电位（Δψm）的增加，它会可逆地改变颜色，从绿色（J-单体，在胞质溶胶中浓度较低时）变为红色（J-聚集体，在活跃线粒体中浓度较高时）。我们表明，除了Δψm的变化外，这种特定染料还可用于追踪线粒体分布和线粒体网络连通性的改变。我们认为JC-1是用于比较不同治疗组的更优探针，因为绿色与红色荧光强度的比值用于分析。该比值仅取决于线粒体膜电位，而不取决于其他线粒体相关或无关因素。我们展示了JC-1染色在研究精神分裂症患者和健康受试者来源的不同细胞类型中线粒体异常方面的各种应用。
