The mitochondrial respiratory activity of spermatozoa was evaluated with a polarographic assay of oxygen consumption performed in sperm cells. Polarography measures the rate of change of oxygen concentration in a solution and, since oxygen is the last in the respiratory chain to accept electrons, provides a direct measure of mitochondrial activity [38 (link)].
Oxygen uptake by rat spermatozoa (2 × 107 sperm cells) was measured by using a Clark-type oxygen probe (Hansatech oxygraph; Hansatech Pentney, King’s Lynn, UK). Oxygen consumption was measured in the presence of a solution of respiratory substrates (10 mmol/L pyruvate and 10 mmol/L malate) and 0.76 μmol/L of adenosine diphosphate (ADP) (107360, 240176 and A2754, Sigma Aldrich).
Sperm mitochondria respiration efficiency was measured using the RCR, which was calculated by dividing rate of oxygen uptake measured in the presence of substrates + ADP (V3) by rate of oxygen uptake measured with substrates alone (V4) [10 (link)].
Oxygen uptake by rat spermatozoa (2 × 107 sperm cells) was measured by using a Clark-type oxygen probe (Hansatech oxygraph; Hansatech Pentney, King’s Lynn, UK). Oxygen consumption was measured in the presence of a solution of respiratory substrates (10 mmol/L pyruvate and 10 mmol/L malate) and 0.76 μmol/L of adenosine diphosphate (ADP) (107360, 240176 and A2754, Sigma Aldrich).
Sperm mitochondria respiration efficiency was measured using the RCR, which was calculated by dividing rate of oxygen uptake measured in the presence of substrates + ADP (V3) by rate of oxygen uptake measured with substrates alone (V4) [10 (link)].
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