V. Yu. Denisenko
All-Russian Research Institute of Genetics and Breeding of Farm Animals, branch of Federal Scientific Center of Animal Husbandry – Ernst All-Russian Research Institute of Animal Husbandry, Moskovskoe sh., 55a, Sankt-Peterburg, 196601, Russian Federation
Abstract. In this study, the effect of protein kinase A and C inhibitors (compounds H-89 and Ro 31-8220, respectively) on the acrosome reaction and the amount of intracellular calcium in bovine spermatozoa was studied using chlortetracycline (CTC) fluorescence assay. The work was carried out in 2019 on the ejaculate of Black-and-White and Irish bulls. In the presence of progesterone, the separate introduction of guanosine diphosphate (GDP) at a concentration of 50 uM and prolactin (PRL) at a concentration of 10 ng/mL stimulated the release of intracellular Ca2+ in bovine spermatozoa both in the presence of the protein kinase A inhibitor and after the treatment with the protein kinase C inhibitor. The combined action of GDP and PRL activated an additional release of intracellular Ca2+ in the presence of H-89; however, the presence of the Ro 31-8220 compound had an inhibiting effect. Using chlortetracycline (CTC) fluorescence assay, the cells were divided into the following three groups in terms of their functional status: uncapacitated, capacitated and acrosome-reactive spermatozoa. Incubation in the presence of progesterone for 4 hours with a separate addition of either GDP or PRL did not affect the ratio of different cell types, leaving it unchanged. This was noted in the presence of both H-89 and Ro 31-8220. Under the action of the protein kinase C inhibitor, co-incubation of GDP and PRL for 4 hours resulted in a decrease in the number of acrosome-reactive cells and an increase in the number of capacitated sperm cells. At the same time, the treatment of cells with the protein kinase A inhibitor did not change the ratio of spermatozoa with a different functional status. The protein kinase C was shown to mediate an increase in the acrosome-reactive bovine spermatozoa and provide an additional release of intracellular Ca2+ during cell incubation in the presence of GDP and PRL.
Keywords: protein kinase inhibitors; acrosome reaction; intracellular calcium; bovine spermatozoa.
Author Details: V. Yu. Denisenko, D. Sc. (Biol.), leading research fellow (e-mail: Этот адрес электронной почты защищён от спам-ботов. У вас должен быть включен JavaScript для просмотра.).
For citation: Denisenko VYu. [Effects of protein kinase inhibitors on the acrosome reaction and the concentration of intracellular Ca2+ in bovine spermatozoa]. Dostizheniya nauki i tekhniki APK. 2020;34(5):65-8. Russian. doi: 10.24411/0235-2451-2020-10513.