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可还原迁移型荧光探针用于精确动态评估细胞线粒体膜电位。

Reversibly Migratable Fluorescent Probe for Precise and Dynamic Evaluation of Cell Mitochondrial Membrane Potentials.

机构信息

Institute of Biomedicine and Biotechnology, Shenzhen Institute of Advanced Technology, Chinese Academy of Sciences, Shenzhen 518055, China.

University of Chinese Academy of Sciences, Beijing 100049, China.

出版信息

Biosensors (Basel). 2022 Sep 27;12(10):798. doi: 10.3390/bios12100798.

Abstract

The mitochondrial membrane potential (MMP, ΔΨ) provides the charge gradient required for mitochondrial functions and is a key indicator of cellular health. The changes in MMP are closely related to diseases and the monitoring of MMP is thus vital for pathological study and drug development. However, most of the current fluorescent probes for MMP rely solely on the cell fluorescence intensity and are thus restricted by poor photostability, rendering them not suitable for long-term dynamic monitoring of MMP. Herein, an MMP-responsive fluorescent probe pyrrolyl quinolinium () which is capable of reversible migration between mitochondria and nucleolus is developed and demonstrated for dynamic evaluation of MMP. The fluorescence of translocates from mitochondria to nucleoli when MMP decreases due to the intrinsic RNA-specificity and more importantly, the translocation is reversible. The cytoplasm to nucleolus fluorescence intensity ratio is positively correlated with MMP so that this method avoids the negative influence of photostability and imaging parameters. Various situations of MMP can be monitored in real time even without controls. Additionally, long-term dynamic evaluation of MMP is demonstrated for HeLa cells using in oxidative environment. This study is expected to give impetus to the development of mitochondria-related disease diagnosis and drug screening.

摘要

线粒体膜电位 (MMP,ΔΨ) 提供了线粒体功能所需的电荷梯度,是细胞健康的关键指标。MMP 的变化与疾病密切相关,因此监测 MMP 对于病理研究和药物开发至关重要。然而,目前大多数用于 MMP 的荧光探针仅依赖于细胞荧光强度,因此受到较差的光稳定性的限制,不适合 MMP 的长期动态监测。在此,我们开发了一种 MMP 响应型荧光探针吡咯并喹啉鎓 (),它能够在线粒体和核仁之间可逆迁移,用于 MMP 的动态评估。当由于内在的 RNA 特异性导致 MMP 降低时,的荧光从线粒体转移到核仁,更重要的是,这种迁移是可逆的。细胞质到核仁的荧光强度比与 MMP 呈正相关,因此该方法避免了光稳定性和成像参数的负面影响。甚至无需对照即可实时监测各种 MMP 情况。此外,我们使用 在氧化环境中对 HeLa 细胞进行了 MMP 的长期动态评估。这项研究有望为线粒体相关疾病的诊断和药物筛选的发展提供动力。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6c89/9599583/d05f38e96838/biosensors-12-00798-g001.jpg

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