DoBC:Methods

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Major Methods Used In The Department

The experiments are performed on living rats, isolated segments of the vessels and the heart of rats, as well as on the endothelium and the mitochondria from isolated rat hearts.

Methods Used In Experiments

Physiological methods: registration of cardiohemodynamics; retrograde perfusion of coronary artery by the Langendorf method, recording of oxygen tension in solutions, registration of active myogenic responses of isolated vascular smooth muscle preparations, registration of electrical responses of the endothelium, injections of substances in populations of brain neurons using stereotaxic coordinates;

biophysical methods: recording of membrane potential of endothelial cells (cultivated and intact) by perforated patch -clamp under current clamp mode;

biochemical methods: determination of hydrogen sulfide, activities of hydrogen sulfide producing enzymes, activities of nitric oxide synthases (neuronal, endothelial and inducible), the levels of indicators of oxidative and nitrosative stress (nitrate, nitrite, nitrosothiols, peroxide hydrogen, superoxide radicals, hydroxyl radicals, diene conjugates, malondialdehyde) in homogenates of heart, aorta, and medulla oblongata, and in suspension of mitochondria, process of mitochondrial respiration and oxidative phosphorylation in the heart mitochondria of rats, isolation of mitochondria from rat heart, spectrophotometric registration of mitochondrial swelling and the mitochondrial factor release; molecular methods: PCR in real time; Western-blot analysis to determine the expression of uncoupling proteins (UCP2 and UCP3) in the mitochondria of the heart

models used: experimental Parkinson’s disease, experimental diabetes mellitus in rats.