Balbach Melanie, Buck Jochen, Levin Lonny R
Department of Pharmacology, Weill Cornell Medical College;
Department of Pharmacology, Weill Cornell Medical College.
J Vis Exp. 2020 Jan 22(155). doi: 10.3791/60815.
Mammalian sperm acquire fertilization capacity in the female reproductive tract in a process known as capacitation. Capacitation-associated processes require energy. There remains an ongoing debate about the sources generating the ATP which fuels sperm progressive motility, capacitation, hyperactivation, and acrosome reaction. Here, we describe the application of an extracellular flux analyzer as a tool to analyze changes in energy metabolism during mouse sperm capacitation. Using H- and O2- sensitive fluorophores, this method allows monitoring glycolysis and oxidative phosphorylation in real-time in non-capacitated versus capacitating sperm. Using this assay in the presence of different energy substrates and/or pharmacological activators and/or inhibitors can provide important insights into the contribution of different metabolic pathways and the intersection between signaling cascades and metabolism during sperm capacitation.
哺乳动物的精子在雌性生殖道中通过一个称为获能的过程获得受精能力。与获能相关的过程需要能量。关于产生为精子的渐进性运动、获能、超活化和顶体反应提供动力的ATP的来源，目前仍存在争议。在这里，我们描述了一种细胞外通量分析仪作为分析小鼠精子获能过程中能量代谢变化的工具的应用。使用对H和O2敏感的荧光团，该方法可以实时监测未获能精子与获能精子中的糖酵解和氧化磷酸化。在存在不同能量底物和/或药理激活剂和/或抑制剂的情况下使用该测定法，可以深入了解不同代谢途径的贡献以及精子获能过程中信号级联与代谢之间的交叉点。
