Published June 15, 2015 | Version v1
Journal article Open

OPTIMIZATION OF SPECIFIC α-TOCOPHEROL CONCENTRATIONS FOR IN VITRO CULTURES OF BOVINE SPERMATOZOA

Creators

  • 1. Department of Animal Physiology, Faculty of Biotechnology and Food Sciences, Slovak University of Agriculture, Nitra, Slovakia

Description

This study was designed to assess the dose- and time-dependent in vitro effects of α-tocopherol (α-TOC) on bovine spermatozoa during different time periods (Times 0h, 2h, 6h, 12h, 18h, 24h). Semen samples were collected from 20 adult breeding bulls, and diluted in physiological saline solution containing 0.5% ethanol (96%) together with 0, 1, 5, 10, 50, 100, 200, 500, 1000 and 2000 μM/L of α-TOC. Most α-TOC concentrations had beneficial effects on the spermatozoa motion parameters, while groups supplemented with 500 and 200 μM/L exhibiting significant differences (P<0.05 and P<0.001, respectively). The MTT assay indicated that none of the α-TOC concentrations had a negative or cytotoxic effect on the spermatozoa mitochondrial activity, and moreover showed a significantly (P<0.05; P<0.01; P<0.001, respectively) improved cell viability in all the experimental groups throughout the in vitro culture. The NBT test showed that the addition of 1000-50 μM/L α-TOC had an instant positive effect on the spermatozoa protection against free radical production. This protection remained present with a significant impact at all timeframes (P<0.001 and P<0.01, respectively). At the same time, all α-TOC concentrations exhibited significant (P<0.05, P<0.01 and P<0.001, respectively) protective effects on the spermatozoa free radical formation during the end-stages of spermatozoa culture. The results indicate that the addition of α-tocopherol, especially in concentrations ranging between 500 and 50 μM to the culture medium could be beneficial for the overall stimulation of spermatozoa activity and protection against possible in vitro oxidative stress development.

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