Detection of the Mitochondrial Membrane Potential by the Cationic Dye JC-1 in L1210 Cells with Massive Overexpression of the Plasma Membrane ABCB1 Drug Transporter.
Detection of the Mitochondrial Membrane Potential by the Cationic Dye JC-1 in L1210 Cells with Massive Overexpression of the Plasma Membrane ABCB1 Drug Transporter.
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DOI：
10.3390/ijms19071985
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发表时间：
2018-07-07
影响因子：
5.6
通讯作者：
Breier A
中科院分区：
文献类型：
--
作者：
Elefantova K;Lakatos B;Kubickova J;Sulova Z;Breier A
JC-1是一种阳离子荧光染料，当添加到活细胞中时，已知其仅定位于线粒体中，特别是在以足够的线粒体膜电位（Δ λ）为特征的良好生理条件下。JC-1在这些细胞器中的积累导致形成J-聚集体（在590 nm处具有特定的红色荧光发射最大值），这是除了J-单体的典型绿色荧光（在529 nm处的发射最大值）之外的。线粒体Δ ε的缺乏导致JC-1线粒体积累的抑制和J-聚集体形成的减少。因此，负载JC-1的细胞的红色和绿色荧光之间的比率经常用于检测线粒体膜电位。然而，JC-1代表多药物转运蛋白P-糖蛋白（P-gp）的合适底物。因此，在P-gp阳性细胞中，可以预期胞内空间中，特别是线粒体中的JC-1含量降低到J聚集体形成无效的水平。在本文中，我们在亲本P-gp阴性L1210（S）细胞及其通过长春新碱（R）选择或转染编码P-gp的人基因（T）获得的P-gp阳性变体上证明了这一行为。P-糖蛋白抑制剂环孢菌素A和维拉帕米不能恢复JC-1加载的R和T细胞的程度类似于在S细胞中观察到的。相比之下，非竞争性高亲和力P-gp抑制剂tariquidar完全恢复了JC-1的积累和J-聚集体典型红色荧光的存在。在存在tariquidar的情况下，JC-1荧光的测量揭示了P-gp阴性（S）和P-gp阳性细胞（R和T）中相似水平的线粒体膜电位。
JC-1, a cationic fluorescent dye when added to living cells, is known to be localized exclusively in mitochondria, particularly in good physiological conditions characterized by sufficient mitochondrial membrane potential (ΔΨ). The accumulation of JC-1 in these organelles leads to the formation J-aggregates (with a specific red fluorescence emission maximum at 590 nm), which is in addition to the typical green fluorescence of J-monomers (emission maximum of ∼529 nm). The lack of mitochondrial ΔΨ leads to the depression of JC-1 mitochondrial accumulation and a decrease in J-aggregate formation. Therefore, the ratio between the red and green fluorescence of cells loaded with JC-1 is often used for the detection of the mitochondrial membrane potential. However, JC-1 represents a suitable substrate of the multidrug transporter P-glycoprotein (P-gp). Therefore, the depression of the JC-1 content in intracellular space and particularly in the mitochondria to a level that is inefficient for J-aggregate formation could be expected in P-gp-positive cells. In the current paper, we proved this behavior on parental P-gp-negative L1210 (S) cells and their P-gp-positive variants obtained by either selection with vincristine (R) or transfection with the human gene encoding P-gp (T). P-glycoprotein inhibitors cyclosporine A and verapamil fail to restore JC-1 loading of the R and T cells to an extent similar to that observed in S cells. In contrast, the noncompetitive high affinity P-gp inhibitor tariquidar fully restored JC-1 accumulation and the presence of the typical red fluorescence of J-aggregates. In the presence of tariquidar, measurement of the JC-1 fluorescence revealed similar levels of mitochondrial membrane potential in P-gp-negative (S) and P-gp-positive cells (R and T).
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影响因子：
9.3
作者：
Kosztyu, Petr;Dolezel, Petr;Mlejnek, Petr
通讯作者：
Mlejnek, Petr
影响因子：
3
作者：
Breier, A.;Stetka, J.;Sulova, Z.
通讯作者：
Sulova, Z.
影响因子：
5.1
作者：
Mlejnek, Petr;Kosztyu, Petr;Ruzickova, Eliska
通讯作者：
Ruzickova, Eliska
影响因子：
5.8
作者：
Ortiz-Lazareno PC;Bravo-Cuellar A;Lerma-Díaz JM;Jave-Suárez LF;Aguilar-Lemarroy A;Domínguez-Rodríguez JR;González-Ramella O;De Célis R;Gómez-Lomelí P;Hernández-Flores G
通讯作者：
Hernández-Flores G
DOI：
10.2147/dddt.s134529
发表时间：
2017
期刊：
Drug design, development and therapy
影响因子：
--
作者：
Du Y;Chen B
通讯作者：
Chen B