NMR spectroscopy of live human asthenozoospermic and normozoospermic sperm metabolism.
NMR spectroscopy of live human asthenozoospermic and normozoospermic sperm metabolism.
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DOI：
10.1530/raf-21-0101
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发表时间：
2022-04-01
影响因子：
2.1
通讯作者：
Pacey, Allan A.
中科院分区：
文献类型：
--
作者：
Reynolds, Steven;Calvert, Sarah J.;Walters, Stephen J.;Paley, Martyn N.;Pacey, Allan A.
不同男性和个别男性的精液之间的精子活力有所不同。我们研究了密度梯度离心洗涤（DCG，80/40%）后正常精子和弱精子的射精，并比较了同一样本中高活力（80%）和低活力（40%）的精子群。我们的目标是确定与精子活力相关的内源代谢组和能量代谢的差异。 1H-核磁共振波谱 (NMR) 测量活体人类精子的内源代谢组。将精子与 13C 标记底物一起孵育，通过 13C-NMR 检测能量代谢。该研究检查了 850 次射精，诊断出弱精子症的比例为 6.1%。使用 DGC 清洗 160 份正常精子 (N) 和 52 份弱精子 (A) 精液，从沉淀中回收高活力精子 (80N/80A)，从界面回收低活力精子 (40N/40A)。 1H-NMR 光谱 45(N) 和 15(A) 被合并，积分通过精子浓度标准化。将来自 126(N) 和 36(A) 精液的精子与 13C-葡萄糖、13C-果糖或 13C-丙酮酸盐一起孵育。 13C-NMR 乳酸和碳酸氢盐积分通过活动精子或活精子浓度进行标准化。 80A 群体的 1H-NMR 谱胆碱积分显着低于 80N，P < 0.0001。当通过活动精子浓度标准化时，40A 精子的 13C 底物转化为乳酸的速度显着高于 80A 精子。偶尔观察到碳酸氢盐积分。弱精子症精液中的精子与正常精子症的精子具有相似的糖酵解需求，但在 40% 和 80% 的精子群体之间观察到更大的差异。 40% 的精子产生的乳酸水平较高可能表明精子活力受损是由于能量代谢失调造成的。胆碱代谢的改变为了解弱精子症的病因提供了机会。不同男性的精液之间精子游动（活力）的差异如何？正常的精子活力有利于受孕，一些被诊断为不孕症的男性精子活力较低。精子代谢分子以产生运动所需的能量。我们测量了精子内分子的浓度和精子分子的代谢，并将这些与活动精子的比例联系起来。该研究检查了 850 个精子样本，发现 6.1% 的精子活力较低。低活力精子样本和正常精子样本之间给予精子的分子代谢相似。然而，当最活跃的精子与其他精子分离时，它们可以更有效地代谢这些分子以实现活力。与正常样本相比，低活力精子样本中的胆碱分子浓度较低。胆碱与细胞膜、能量代谢和氧化应激有关，这可能为了解低活力的原因提供了机会。
Sperm motility varies between ejaculates from different men and from individual men. We studied normozoospermic and asthenozoospermic ejaculates after density-gradient centrifugation washing (DCG, 80/40%) and compared high- (80%) and low (40%)-motility sperm populations within the same sample. Our objective was to identify differences in endogenous metabolomes and energy metabolism in relation to sperm motility. 1H-Nuclear Magnetic Resonance spectroscopy (NMR) measured the endogenous metabolome of live human sperm. Incubating sperm with 13C-labelled substrates detected energy metabolism by 13C-NMR. The study examined 850 ejaculates and diagnosed asthenozoospermia in 6.1%. DGC was used to wash 160 normozoospermic (N) and 52 asthenozoospermic (A) ejaculates to recover high-motility sperm from the pellet (80N/80A) and low motility from the interface (40N/40A). 1H-NMR spectra, 45(N) and 15(A), were binned and the integrals normalised by sperm concentration. Sperm from 126(N) and 36(A) ejaculates were incubated with either 13C-glucose, 13C-fructose or 13C-pyruvate. 13C-NMR lactate and bicarbonate integrals were normalised by motile or vital sperm concentrations. 1H-NMR spectra choline integrals from the 80A population were significantly lower than the 80N, P < 0.0001. 13C-substrate conversion to lactate was significantly higher for 40A sperm than 80A sperm when normalised by motile sperm concentration. Bicarbonate integrals were sporadically observed. Sperm from asthenozoospermic ejaculates had similar glycolytic requirements to normozoospermic ones, with larger differences observed between 40 and 80% sperm populations. Higher lactate levels produced by 40% sperm may indicate that impaired sperm motility is due to dysregulated energy metabolism. The alteration in choline metabolism provides opportunities to understand the aetiology of asthenozoospermia. How well sperm swim (motility) varies between ejaculates from different men? Normal sperm motility is beneficial to conception and some men diagnosed with infertility have low sperm motility. Sperm metabolise molecules to produce the energy required for motility. We measured concentrations of molecules within sperm and metabolism of molecules given to sperm and related these to the proportion of motile sperm. The study examined 850 sperm samples and found low motility in 6.1%. Metabolism of molecules given to sperm was similar between low and normal motility sperm samples. However, when the most motile sperm were separated from the rest, they were more efficient in metabolising these molecules to achieve motility. Lower concentrations of a molecule called choline were found in low-motility sperm samples compared to normal samples. Choline is associated with cell membranes, energy metabolism and oxidative stress, which may give opportunities to understand the causes of low motility.