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An impedimetric assay for the identification of abnormal mitochondrial dynamics in living cells
Electrophoresis ( IF 3.0 ) Pub Date : 2020-11-09 , DOI: 10.1002/elps.202000125
Kalpani N U Galpayage Dona 1 , E Du 2 , Jianning Wei 3
Affiliation  

Mitochondrial dynamics (fission and fusion) plays an important role in cell functions. Disruption in mitochondrial dynamics has been associated with diseases such as neurobiological disorders and cardiovascular diseases. Analysis of mitochondrial fission/fusion has been mostly achieved through direct visualization of the fission/fusion events in live‐cell imaging of fluorescently labeled mitochondria. In this study, we demonstrated a label‐free, non‐invasive Electrical Impedance Spectroscopy (EIS) approach to analyze mitochondrial dynamics in a genetically modified human neuroblastoma SH‐SY5Y cell line with no huntingtin protein expression. Huntingtin protein has been shown to regulate mitochondria dynamics. We performed EIS studies on normal SH‐SY5Y cells and two independent clones of huntingtin‐null cells. The impedance data was used to determine the suspension conductivity and further cytoplasmic conductivity and relate to the abnormal mitochondrial dynamics. For instance, the cytoplasm conductivity value was increased by 11% from huntingtin‐null cells to normal cells. Results of this study demonstrated that EIS is sensitive to characterize the abnormal mitochondrial dynamics that can be difficult to quantify by the conventional microscopic method.

中文翻译:

用于鉴定活细胞中异常线粒体动力学的阻抗测定法

线粒体动力学(裂变和融合)在细胞功能中起着重要作用。线粒体动力学的破坏与神经生物学障碍和心血管疾病等疾病有关。线粒体裂变/融合的分析主要是通过荧光标记线粒体活细胞成像中裂变/融合事件的直接可视化来实现的。在这项研究中,我们展示了一种无标记、非侵入性的电阻抗光谱 (EIS) 方法来分析没有亨廷顿蛋白表达的转基因人类神经母细胞瘤 SH-SY5Y 细胞系的线粒体动力学。亨廷顿蛋白已被证明可以调节线粒体动力学。我们对正常的 SH-SY5Y 细胞和亨廷顿蛋白无效细胞的两个独立克隆进行了 EIS 研究。阻抗数据用于确定悬浮液电导率和进一步的细胞质电导率,并与异常线粒体动力学有关。例如,从亨廷顿蛋白无效细胞到正常细胞,细胞质电导率值增加了 11%。这项研究的结果表明,EIS 对表征异常线粒体动力学很敏感,而这些异常线粒体动力学很难通过传统的显微方法量化。
更新日期:2021-01-14
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