The effect of testosterone on Сa²⁺ release stimulated by somatotropin and theophylline from intracellular stores of <i>Sus Scrofa Domesticus</i> oocytes

Abstract

Purpose. Studying the influence of testosterone on stimulated by somatotropin and theophylline liberation of Ca2+ from intracellular depot of oocytes of pigs.Materials and methods. The material for the studies was oocytes secreted from antral follicles (with a diameter of 3-6 mm) of the ovarian Sus Scrofa domesticus. Oocyte complexes were aspirated from the ovaries at the stage of follicular growth, without signs of visible pathology. The dedicated oocytes were incubated in the modified incubation environment Dulbekko without CaCl2, containing 36 mg/l of Piruvat NA and 1 g/l glucose. Caicium in the intracellular depot of oocytes of pigs was measured with the help of a chlortetracycline (CTC) fluorescent probe. Oocytes were loaded with a probe for 5 minutes at 370C in an environment containing 40 microns of CTC. Then the cells were washed three times in an incubation environment and transferred to a special quartz glass with cells of 0.05 ml. Dependent on Ca2+ fluorescence of the CTC was recorded in oocytes in the environment of Dulbekko. The intensity of the fluorescence of the CTC probe was measured on a fluorimetric installation consisting of a fluorescent microscope, equipped with the necessary light filters and a photometric nozzle of the FMEL-1A. The CTC-Ca2+complex-the membrane excited 380-400 nm light, fluorescence was recorded in the area of 530 nm. The intensity of fluorescence was measured in the conc. units. The duration of ultraviolet radiation on oocytes during measurements did not exceed 5 seconds. In all experiments, an EGT was added to the incubation environment at a concentration of 0.5 mm.Results. It was shown that in the absence of testosterone in oocytes, the addition of somatotropin (bST) or theophylline stimulated the release of Ca2+ from intracellular depot, while their joint action did not lead to an additional exit of Ca2+ from intracellular depot. Inhibition of proteinkinase and did not affect the liberation of Ca2+, stimulated separately by bST or Theophylline, as well as their joint action. Against the background of the use of testosterone, the addition of bST or theophylline separately did not lead to the release of Ca2+ from intracellular depot. With the joint action of bST and Theophylline in the presence of testosterone, the liberation of Ca2+ from intracellular depot was noted, and the value of this indicator was higher than with the joint action of bST and Theophylline in the absence of testosterone. In stimulated by the joint action of bST and theophylline, the release of Ca2+ from the intracellular depot of oocytes in the presence of testosterone is participated in proteinquinase A and microfilaments, since when exposed to proteinquine A and polymerization of cytochalazine microfilaments release of Ca2+ from intracellular depots was not recorded.