Evaluation of the Integrity of the Respiratory Chain in Rooster Sperm after Freezing

Abstract

One of the causes of cell death is a violation of electron transfer through the respiratory chain of mitochondria. The aim of study was to assess the integrity of the respiratory chain sperm after freezing in roosters. The effect of 2.4 dinitrophenol (2.4 DNP) on NADH fluorescence in fresh and cryopreserved semen of roosters was investigated. If the transfer of electrons and protons along the respiratory chain functions normally 2.4 DNP enhances the nonphosphorylating oxidation of NADH and its fluorescence decreases. The sperm of 30 cocks was frozen. The NADH was measured on a LUMAM fluorescence microscope. The luminescence excitation wavelength of NADH is 365 nm. Registration of NADH was carried out at a wavelength 465 nm. The amount of NADH was expressed in conventional units and the change in fluorescence after addition of 2.4 DNP was calculated as a percentage of the initial value taken as 100 %. The fluorescence of NADH after adding 2.4 DNP to frozen semen was 9% less than in fresh semen. A relationship between the magnitude of the change in NADH fluorescence after the addition of DNP and egg fertilization was established (r =0.65; P<0.05). Respiratory stimulation by 2.4 DNP is an useful and informative indicator in assessing sperm quality. Support or Funding Information The study was supported by the Russian Science Foundation (project No. 18‐16‐00071)