Inhibition of Mitochondrial Complex I Leads to Decreased Motility and Membrane Integrity Related to Increased Hydrogen Peroxide and Reduced ATP Production, while the Inhibition of Glycolysis Has Less Impact on Sperm Motility.
Inhibition of Mitochondrial Complex I Leads to Decreased Motility and Membrane Integrity Related to Increased Hydrogen Peroxide and Reduced ATP Production, while the Inhibition of Glycolysis Has Less Impact on Sperm Motility.
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DOI：
10.1371/journal.pone.0138777
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发表时间：
2015
期刊：
影响因子：
3.7
通讯作者：
Peña FJ
中科院分区：
文献类型：
--
作者：
Plaza Davila M;Martin Muñoz P;Tapia JA;Ortega Ferrusola C;Balao da Silva C C;Peña FJ
线粒体被认为是体细胞和精子中活性氧的主要来源。然而，没有关于线粒体活性氧在种马精子中的作用的数据。为了阐明线粒体电子传递链在种马精子氧化应激起源中的作用，使用了复合物I（鱼藤酮）和III（抗霉素A）的特异性抑制剂。收集来自七匹安达卢西亚种马的射精，并在鱼藤酮、抗霉素-A或对照媒介物存在下在37°C下在BWW培养基中孵育。在这些抑制剂的存在下孵育降低精子活力和速度（CASA分析）（p<0.01），但在鱼藤酮（复合物I抑制剂）的存在下效果更明显。这些抑制剂也降低ATP含量。复合物I和III的抑制降低了活性氧物质的产生（p<0.01），如用CellRox深红染色后通过流式细胞术评估的。这一观察结果表明，CellRox探针主要识别超氧化物，超氧化物的产生可能反映了强烈的线粒体活性，而不是氧化应激。复合物I的抑制导致过氧化氢产生增加（p<0.01）。糖酵解的抑制导致精子速度降低（p<0.01），而对总活动精子的百分比没有影响。弱和中度（但有统计学意义）的正相关性之间观察到精子活力，速度和膜完整性和活性氧的产生。这些结果表明，种马精子在很大程度上依赖于氧化磷酸化（OXPHOS）生产ATP的运动，但也需要糖酵解，以保持高速度。这些数据还表明，增加过氧化氢起源于线粒体是一种机制参与种马精子衰老。
Mitochondria have been proposed as the major source of reactive oxygen species in somatic cells and human spermatozoa. However, no data regarding the role of mitochondrial ROS production in stallion spermatozoa are available. To shed light on the role of the mitochondrial electron transport chain in the origin of oxidative stress in stallion spermatozoa, specific inhibitors of complex I (rotenone) and III (antimycin-A) were used. Ejaculates from seven Andalusian stallions were collected and incubated in BWW media at 37°C in the presence of rotenone, antimycin-A or control vehicle. Incubation in the presence of these inhibitors reduced sperm motility and velocity (CASA analysis) (p<0.01), but the effect was more evident in the presence of rotenone (a complex I inhibitor). These inhibitors also decreased ATP content. The inhibition of complexes I and III decreased the production of reactive oxygen species (p<0.01) as assessed by flow cytometry after staining with CellRox deep red. This observation suggests that the CellRox probe mainly identifies superoxide and that superoxide production may reflect intense mitochondrial activity rather than oxidative stress. The inhibition of complex I resulted in increased hydrogen peroxide production (p<0.01). The inhibition of glycolysis resulted in reduced sperm velocities (p<0.01) without an effect on the percentage of total motile sperm. Weak and moderate (but statistically significant) positive correlations were observed between sperm motility, velocity and membrane integrity and the production of reactive oxygen species. These results indicate that stallion sperm rely heavily on oxidative phosphorylation (OXPHOS) for the production of ATP for motility but also require glycolysis to maintain high velocities. These data also indicate that increased hydrogen peroxide originating in the mitochondria is a mechanism involved in stallion sperm senescence.