Induction of capacitation of bovine spermatozoa before cryopreservation increases their viability after thawing
- Authors: Denisenko V.Y.1, Kuzmina T.I1, Boytseva E.N1
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Affiliations:
- Russian Research Institute of Farm Animal Genetics and Breeding - Branch of the L.K. Ernst Federal Science Center for Animal Husbandry
- Issue: Vol 13, No 2 (2018)
- Pages: 72-76
- Section: Articles
- URL: https://genescells.ru/2313-1829/article/view/120748
- DOI: https://doi.org/10.23868/201808023
- ID: 120748
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Abstract
Cryopreservation of sperm is an important tool of reproductive biotechnology in the solving of the problems in infertility and reproduction of animals. In despite of the achievements in this field, the mechanisms that determine the cryoresistance of male gametes require further study. The damaging effect of ultralow temperatures during cryopreservation is primarily directed to the plasma membrane of spermatozoa. The purpose of this study is to analyze the viability of thawed bull spermatozoa after preventively induced capacitation with the further cryopreservation. Ejaculates of three noninbred bulls of Ayrshire and black-and-white breeds were used in the experiments. The functional state of spermatozoa was evaluated with a chlortetracycline test. Gametes were ranked in accordance with one of the three types of fluorescence of CTC-calcium-membrane complex : uniform fluorescence throughout the head (uncapacitated cells); fluorescence-free band in the post-acrosome region (capacitated cells); low fluorescence in the entire head, except for a thin bright fluorescence band in the equatorial segment (acrosome-reactive cells). The viability of spermatozoa was assessed with propidium iodide (5 |jg / ml). Capacitation was induced by heparin (5 jg / ml) or theophylline / dbcAMP (250/100 jM). It was shown that the induction of the capacitation of bull sperm by heparin, as well as theophylline /dbcAMP before cryopreservation enhances the number of viable sperm, reduces the number of capacitated cells and increases the number of sperm with acrosome reaction after thawing. In intact (without freezing) spermatozoa after induction of capacitation such effect is not revealed, the number of viable cells remained unchanged, the induction of capacitation caused an increase number of capacitated spermatozoa. The findings expand knowledge concerning the effects of extremely low temperatures on male gametes and can be used for modernization of the cryopreservation technology for increase their survival rate after thawing.
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About the authors
V. Yu Denisenko
Russian Research Institute of Farm Animal Genetics and Breeding - Branch of the L.K. Ernst Federal Science Center for Animal Husbandry
T. I Kuzmina
Russian Research Institute of Farm Animal Genetics and Breeding - Branch of the L.K. Ernst Federal Science Center for Animal Husbandry
Email: prof.kouzmina@mail.ru
E. N Boytseva
Russian Research Institute of Farm Animal Genetics and Breeding - Branch of the L.K. Ernst Federal Science Center for Animal Husbandry
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