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Thioflavin T indicates mitochondrial membrane potential in mammalian cells
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Skates, Emily, Delattre, Hadrien, Schofield, Zoe, Asally, Munehiro and Soyer, Orkun S. (2023) Thioflavin T indicates mitochondrial membrane potential in mammalian cells. Biophysical Reports, 3 (4). 100134. doi:10.1016/j.bpr.2023.100134 ISSN 2667-0747.
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Official URL: https://doi.org/10.1016/j.bpr.2023.100134
Abstract
The fluorescent benzothiazole dye thioflavin T (ThT) is widely used as a marker for protein aggregates, most commonly in the context of neurodegenerative disease research and diagnosis. Recently, this same dye was shown to indicate membrane potential in bacteria due to its cationic nature. This finding prompted a question whether ThT fluorescence is linked to the membrane potential in mammalian cells, which would be important for appropriate utilization of ThT in research and diagnosis. Here, we show that ThT localizes into the mitochondria of HeLa cells in a membrane-potential-dependent manner. Specifically, ThT colocalized in cells with the mitochondrial membrane potential indicator tetramethylrhodamine methyl ester (TMRM) and gave similar temporal responses as TMRM to treatment with a protonophore, carbonyl cyanide-4-(trifluoromethoxy) phenylhydrazone (FCCP). Additionally, we found that presence of ThT together with exposure to blue light (λ = 405 nm), but neither factor alone, caused depolarization of mitochondrial membrane potential. This additive effect of the concentration and blue light was recapitulated by a mathematical model implementing the potential-dependent distribution of ThT and its effect on mitochondrial membrane potential through photosensitization. These results show that ThT can act as a mitochondrial membrane potential indicator in mammalian cells, when used at low concentrations and with low blue light exposure. However, it causes dissipation of the mitochondrial membrane potential depending additively on its concentrations and blue light exposure. This conclusion motivates a re-evaluation of ThT’s use at micromolar range in live-cell analyses and indicates that this dye can enable future studies on the potential connections between mitochondrial membrane potential dynamics and protein aggregation.
Item Type: | Journal Article | |||||||||||||||
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Subjects: | Q Science > QD Chemistry Q Science > QH Natural history Q Science > QP Physiology |
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Divisions: | Faculty of Science, Engineering and Medicine > Science > Life Sciences (2010- ) | |||||||||||||||
SWORD Depositor: | Library Publications Router | |||||||||||||||
Library of Congress Subject Headings (LCSH): | Benzothiazoles, Mitochondrial membranes, Animal cell biotechnology, Mammals, Proteins | |||||||||||||||
Journal or Publication Title: | Biophysical Reports | |||||||||||||||
Publisher: | Elsevier | |||||||||||||||
ISSN: | 2667-0747 | |||||||||||||||
Official Date: | 13 December 2023 | |||||||||||||||
Dates: |
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Volume: | 3 | |||||||||||||||
Number: | 4 | |||||||||||||||
Article Number: | 100134 | |||||||||||||||
DOI: | 10.1016/j.bpr.2023.100134 | |||||||||||||||
Status: | Peer Reviewed | |||||||||||||||
Publication Status: | Published | |||||||||||||||
Access rights to Published version: | Open Access (Creative Commons) | |||||||||||||||
Date of first compliant deposit: | 30 November 2023 | |||||||||||||||
Date of first compliant Open Access: | 30 November 2023 | |||||||||||||||
RIOXX Funder/Project Grant: |
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