The use of the Petri net method in the simulation modeling of mitochondrial swelling.

Using photon correlation spectroscopy, which allows investigating changes in the hydrodynamic dia­meter of the particles in suspension, it was shown that ultrahigh concentrations of Ca2+ (over 10 mM) induce swelling of isolated mitochondria. An increase in hydrodynamic diameter was caused by an increase of non-specific mitochondrial membrane permeability to Ca ions, matrix Ca2+ overload, activation of ATP- and Ca2+-sensitive K+-channels, as well as activation of cyclosporin-sensitive permeability transition pore. To formalize the experimental data and to assess conformity of experimental results with theoretical predictions we developed a simulation model using the hybrid functional Petri net method.

CALIX[4]ARENES AS MODULATORS OF ENERGY-DEPENDENT CA2+_ACCUMULATION AND FUNCTIONING OF THE ELECTRON TRANSPORT CHAIN IN SMOOTH MUSCLE MITOCHONDRIA.

The influence of supramolecular macrocyclic compounds calix[4]arenes (C-97, C-99, C-107) at a concentration of 100 nM in the process of energy-dependent Ca²⁺-transport in isolated mitochondria of smooth muscle, as well as autofluorescence mitochondrial coenzyme NADH, FAD and hydrodynamic diameter of these organelles was investigated. Using Ca²⁺-sensitive fluorescent dye Fluo-4 AM it was shown that the selected calix[4]arenes can suppress energy-dependent accumulation of Ca²⁺ by mitochondria. Accumulation of Ca²⁺ (80 jiM in the medium) accompanied by the growth of the fluorescent probe response from a conventional unit to a value of 1,57±0,04 (n=5).

ELECTROCHEMICAL POTENTIAL OF THE INNER MITOCHONDRIAL MEMBRANE AND Ca2+ HOMEOSTASIS OF MYOMETRIUM CELLS.

We demonstrated using Ca(2+)-sensitive fluorescent probe, mitochondria binding dyes, and confocal laser scanning microscopy, that elimination of electrochemical potential of uterus myocytes' inner mitochondrial membrane by aprotonophore carbonyl cyanide m-chlorophenyl hydrazone (10 μM), and by a respiratory chain complex IV inhibitor sodium azide (1 mM) is associated with substantial increase of Ca2+ concentration in myoplasm in the case of the protonophore effect only, but not in the case of the azide effect. In particular, with the use of nonyl acridine orange, a mitochondria-specific dye, and 9-aminoacridine, an agent that binds to membrane compartments in the presence of proton gradient, we showed that both the protonophore and the respiratory chain inhibitor cause the proton gradient on mitochondrial inner membrane to dissipate when introduced into incubation medium. We also proved with the help of 3,3'-dihexyloxacarbocyanine, a potential-sensitive carbocyanine-derived fluorescent probe, that the application of these substances results in dissipation of the membrane's electrical potential.