The estimation of isolated hepatocytes mitochondrial potential at change of the oxidizing status
Abstract
In the present study, the method of fluorescent probes has been applied for estimation of the changes of the transmembrane potential of mitochondria under the conditions of oxidizing stress. With the help of mitochondrion probe JС–1 it has been shown, that under influence of Н 2 О 2 in conditions of long incubation (within 3 hours) there is a decrease in transmembrane potential which is not suppressed by thiol-reducing connection DTT (dithiothreitol). TBHP (tert-butil-hidroperoxide) also was found to cause no changes in mitochondrion potential under the given experimental conditions. Incubation of the cells with adrenaline was accompanied by an increase in fluorescence intensity of the modular form of the dye. Dynamics of changes of potential at the initial stages of cellular activation was started by adrenaline use as prooxidizer of hydrogen peroxide, however, the character of the answer changed essentially on a background of TBHP action pointing to the various mechanisms of influence of the given compounds on studied parameters.
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References
Freedman J.C., Novak T.S. Membrane potentials associated with Ca-induced K conductance in human
red blood cells: studies with a fluorescent oxonol dye, WW 781//J Membr Biol. -1983. 72, 59-74
Gross D., Loew L.M., Webb W.W. Optical imaging of cell membrane potential changes induced by
applied electric fields//Biophys J.1986. 50. 339-348
Флуоресцентные зонды в исследовании биологических мембран./ Ю.А. Владимиров, Г.Е.
Добрецов – М.: Наука, 1980. - 318 с.
В.П. Скулачев. Явления запрограммированной смерти. Митохондрии, клетки и органы: роль
активных форм кислорода // Соросовский образовательный журнал. 2001. № 6. С.4-10.
Ю.А. Владимиров. Свободные радикалы в живых системах // Соросовский образовательный
журнал. 2000. № 6. С.13-19.
Johnson L.V., Walsh M.L., Chen L.B. Localization of mitochondria in living cells with rodamine
//Proc Natl Acad Sci USA. 1980. 77. 990.
Salvioli S., Ardizzoni A., Franceschi C., Cossarizza A. JC-1, but not DiOC6(3) or rhodamine 123, is a
reliable fluorescent probe to assess ΔΨ changes in intact cells: implication for studies on mitochondrial
functionality during apoptosis//FEBS Letters 1997. P. 77-82.
Diaz G., Setzu M.D., Zucca A., Isola R., Diana A., Murru R., Sogos V., Gremo F. Subcellular
heterogeneity of mitochondrial membrane potential: relationship with organelle distribution and
intercellular contacts in normal, hypoxic and apoptotic cells// Journal of Cell Science 1999.112, 1077-
Diaz G., Falchi A., Gremo F., Isola R., Andrea D. Homogeneous longitudinal pro¢les and synchronous
£uctuations of mitochondrial transmembrane potential//FEBS Letters 2000 475 218-224.
DiLisa F., Blank P.S., Colonna R., Gambassi G., Silverman H.S., Stern M.D., Hansford R.G.
Mitochondrial membrane potential in single living adult rat cardiac myocytes exposed to anoxia or
metabolic inhibition//Journal of physiology 1995. 486, 1. 1-13.
Канаева И.П., Карякин А.В., Аленичева Т.В., Бурмакова Г.А., Алимов Г.А. и др.
//Цитология.1975. Т.17. с 545-551.
Connern C.P., Halestrap A.P. Recruitment of mitochondrial cyclophilin to the mitochondrial inner
membrane under conditions of oxidative stress that enhance the opening of a calcium-sensitive nonspecific channel//Biochem. J. 1994. 302, 321-324.
Imberti R., Nieminen A.L., Herman B., Lemasters J.J. Mitochondrial and glycolytic dysfunction in
lethal injury to hepatocytes by t-butylhydroperoxide: protection by fructose, cyclosporin A and
trifluoperazine //J. Pharmacol. Exper. Ther. 1993.265, 392-400.
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