Garrett Louise J A, Revell Stuart G, Leese Henry J
Department of Biology, University of York, York YO10 5YW, United Kingdom.
J Androl. 2008 Jul-Aug;29(4):449-58. doi: 10.2164/jandrol.107.003533. Epub 2007 Nov 28.
This article's objectives are to investigate the relationship between adenosine triphosphate (ATP) production (oxidative phosphorylation and glycolysis) and fertility of bovine spermatozoa, determine the proportion of oxygen consumption devoted to proton leak and that due to nonmitochondrial processes, and discover whether freeze/thawing affects sperm oxygen consumption. Oxygen consumption of bovine spermatozoa was measured using a standard Clark electrode and, for the first time, in an Oxygen Biosensor System (OBS). Total ATP formation by bovine spermatozoa was calculated from the oxygen consumption and lactate production (glycolysis) by the same spermatozoa sample. ATP production varied from 1.99 to 8.09 mumol ATP per 10(8) spermatozoa per hour; glycolysis accounted for 16% to 38% of ATP. Nonmitochondrial oxygen consumption could not be detected in bovine spermatozoa using these methods. A significant proportion (16%-43%) of oxygen consumption was insensitive to oligomycin and was due to "proton leak." There was no significant difference between oxygen consumption of frozen/thawed and fresh spermatozoa for 2 of the 3 bulls tested. However, oxygen consumption of frozen/thawed spermatozoa was significantly higher (P < .05) than fresh spermatozoa for the third bull. When ZO(2) of frozen/thawed spermatozoa from 20 bulls was compared with their 49 day nonreturn rates (NRRs), oxygen consumption was correlated positively with NRR (ie, fresh spermatozoa with a higher ZO(2) were more fertile). Moreover, total ATP production correlated with NNR better than ZO(2). Bulls with a lower NRR produce spermatozoa that are susceptible to damage during the freeze/thawing process, causing an increase in ZO(2), possibly due to mitochondrial membrane damage resulting in more energy being expended in maintaining the proton gradient, or capacitation-like changes causing hyperactivation. Oxygen consumption measured in the OBS may be useful in assessing bovine sperm fertility.
本文的目的是研究三磷酸腺苷(ATP)生成(氧化磷酸化和糖酵解)与牛精子生育力之间的关系,确定用于质子泄漏和非线粒体过程的耗氧量比例,并探究冻融是否会影响精子耗氧量。使用标准克拉克电极并首次在氧生物传感器系统(OBS)中测量了牛精子的耗氧量。根据同一精子样本的耗氧量和乳酸生成量(糖酵解)计算了牛精子的总ATP生成量。ATP生成量为每10⁸个精子每小时1.99至8.09 μmol ATP;糖酵解占ATP的16%至38%。使用这些方法在牛精子中未检测到非线粒体耗氧量。相当一部分(16% - 43%)的耗氧量对寡霉素不敏感,是由“质子泄漏”导致的。在测试的3头公牛中,有2头的冻融精子和新鲜精子的耗氧量没有显著差异。然而,第三头公牛的冻融精子耗氧量显著高于新鲜精子(P <.05)。当比较20头公牛的冻融精子的Zₒ₂与其49天不返情率(NRR)时,耗氧量与NRR呈正相关(即Zₒ₂较高的新鲜精子生育力更强)。此外,总ATP生成量与NRR的相关性优于Zₒ₂。NRR较低的公牛产生的精子在冻融过程中易受损伤,导致Zₒ₂增加,这可能是由于线粒体膜损伤导致维持质子梯度消耗更多能量,或者是类似获能的变化导致超活化。在OBS中测量的耗氧量可能有助于评估牛精子的生育力。