Journal
JOURNAL OF APPLIED PHYCOLOGY
Volume 31, Issue 6, Pages 3691-3697Publisher
SPRINGER
DOI: 10.1007/s10811-019-01860-3
Keywords
Mitochondrial membrane potential; Chlamydomonas; JC-1; Fluorimetry in vivo; Microplate reader
Funding
- National Science Center, Poland [UMO-2016/23/B/NZ9/00963]
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The fluorochrome JC-1 is mainly used in mammalian cells to estimate mitochondrial membrane potential (MMP) as a stress marker, with far less data being available for plants or green algae. To address this, we have validated the possibility for changes in MMP to be used as a sensitive stress indicator in the green alga Chlamydomonas reinhardtii. To optimize the method, we analyzed the conditions for applicable MMP determination, including the proper buffer, excitation/emission wavelengths, and solvent (dimethyl sulfoxide, DMSO) influence on the fluorescence signal, and evaluated usefulness of the method for toxicological research. Our results have allowed us to develop a complete protocol for the estimation of MMP changes in C. reinhardtii. Statistical analyses have confirmed the reproducibility, sensitivity, and applicability of this method for toxicological studies.
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