The effects of mitochondrial energetics inhibitors on the fluorescence of potential-sensitive dyes rhodamine 123 and DiS-C3-(5) in lymphocyte suspensions |
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Authors: | E N Mokhova I A Rozovskaya |
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Institution: | (1) A. N. Belozersky Laboratory of Molecular Biology and Bioorganic Chemistry, Moscow State University, 119899 Moscow, USSR |
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Abstract: | The effects of uncouplers (FCCP, DNF), oligomycin, and rotenone on the fluorescence of potential-sensitive dyes, rhodamine 123 and diS-C3-(5), in lymphocyte suspensions were compared. The fluorescence of these optical probes gradually increased at higher FCCP concentrations. The dependences of fluorescence intensities and FCCP concentrations were similar for both dyes, and only diS-C3-(5) fluorescence started increasing at lower FCCP concentrations. Rotenone (1 µM) significantly increased rhodamine 123 fluorescence. TMPD-induced and uncoupler-induced diS-C3-(5) fluorescence changes increased 1.5- to 2-fold if the incubation mixture was supplemented with oligomycin (0.1–0.2 µg/ml). The fluorescence responses of the dyes in the lymphocyte suspension correlate with the effects of mitochondrial energetics inhibitors on m in isolated mitochondria. The results suggest the possibility of using these dyes for estimating the direction of the m changes in the lymphocyte suspension.Abbreviations m
difference in electrical potential across the mitochondrial inner membrane
- p
difference in electrical potentials across the plasma membrane
- TMPD
N,N,N,N-tetramethyl-p-phenylenediamine
- DNP
2,4-dinitrophenol
- FCCP
carbonyl cyanidep-trifluoromethoxyphenylhydrazone
- diS-C3-(5)
3,3-dipropylthiodicarbocyanine
- MOPS
morpholinopropane sulfonic acid |
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Keywords: | Lipophilic cation optical probe dis-C3-(5) rhodamine 123 mitochondrial membrane potential uncoupler oligomycin lymphocyte energetics difference spectrophotometer mitochondria in cell |
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