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

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

Abstract

The mitochondrial membrane potential (MMP, ΔΨmito) 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 (PQ) which is capable of reversible migration between mitochondria and nucleolus is developed and demonstrated for dynamic evaluation of MMP. The fluorescence of PQ 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 PQ in oxidative environment. This study is expected to give impetus to the development of mitochondria-related disease diagnosis and drug screening.

Keywords: dynamic evaluation; fluorescent probe; migratable; mitochondrial membrane potential; reversible.

MeSH terms

  • Fluorescent Dyes*
  • HeLa Cells
  • Humans
  • Membrane Potential, Mitochondrial
  • Microscopy, Fluorescence / methods
  • RNA*

Substances

  • Fluorescent Dyes
  • RNA

Grants and funding

This work was financially supported by the National Key R & D Program of China (Grant No. 2020YFA0908800), National Natural Science Foundation of China (Grant Nos. 32000981, 81903057, and 31922040), Shenzhen Science and Technology Research Funding (Grant Nos. JCYJ20190806170011328 and JCYJ20210324102202007), China National Postdoctoral Program for Innovative Talents (Grant No. BX20190352), Guangdong Basic and Applied Basic Research Foundation (Grant No. 2019A1515111209), City University of Hong Kong Donation Research Grant (DON-RMG 9229021), City University of Hong Kong Donation Grant (9220061), Hong Kong PDFS—RGC Postdoctoral Fellowship Scheme (PDFS2122-1S08 and CityU 9061014), Hong Kong ITC (Innovation and Technology Commission) ITF (Innovation and Technology Fund) (GHP/149/20SZ and CityU 9440296), and Hong Kong HMRF (Health and Medical Research Fund) (2120972 and CityU 9211320).