Premium
The mitochondrial membrane potential in human platelets: a sensitive parameter for platelet quality
Author(s) -
Verhoeven Arthur J.,
Verhaar Robin,
Gouwerok Eric G.W.,
De Korte Dirk
Publication year - 2005
Publication title -
transfusion
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.045
H-Index - 132
eISSN - 1537-2995
pISSN - 0041-1132
DOI - 10.1111/j.1537-2995.2005.04023.x
Subject(s) - platelet , membrane potential , flow cytometry , fluorescence , oxidative phosphorylation , mitochondrion , chemistry , biophysics , membrane integrity , membrane , biochemistry , analytical chemistry (journal) , biology , microbiology and biotechnology , chromatography , immunology , physics , quantum mechanics
BACKGROUND: Deterioration of platelet (PLT) quality during storage is accompanied by an increase in lactate production, indicating a decrease in mitochondrial function. In this study, the optimal conditions under which the fluorescent dye JC‐1 can be used to detect changes in mitochondrial function in PLTs were established. STUDY DESIGN AND METHODS: PLTs were incubated at 37°C in synthetic medium under various conditions of JC‐1 loading. In the presence of a high membrane potential, this dye accumulates in the mitochondria with a concomitant increase in red fluorescence. After JC‐1 loading, the ratio of red (FL2) to green (FL1) fluorescence was determined by flow cytometry. RESULTS: The FL2‐to‐FL1 ratio of PLTs (3 × 10 7 /mL, loaded with 0.5 µmol/L JC‐1) amounted to about 5 in 1‐day‐old PLTs. At higher dye concentrations, the FL2‐to‐FL1 ratio was significantly lower, suggesting uncoupling by the dye itself. Plasma concentrations above 3 percent significantly affected the JC‐1 signal. The FL2‐to‐FL1 ratio showed a dose‐dependent decrease to an uncoupler of oxidative phosphorylation or to inhibition of the respiratory chain. JC‐1‐loaded PLTs showed a clear decrease in FL2‐to‐FL1 ratio after prolonged storage or upon ultraviolet (UV) illumination. Only after UV treatment did changes in JC‐1 signal correlate with changes in CD62P expression. CONCLUSION: The FL2‐to‐F1 ratio of PLTs loaded with JC‐1 is a reliable and sensitive indicator of the mitochondrial membrane potential, provided that the proper experimental conditions have been applied.