精液冷冻保存导致精子中存在的分子的差异重塑,这些与精子质量及其生理功能降低有关的改变尚未得到充分理解。鉴于此,本研究通过分析山羊精子在冷冻保存过程中的代谢特征变化,探讨山羊精子冷冻损伤的机制。采用超高效液相色谱-四极杆飞行时间质谱(UHPLC-QTOF-MS)技术研究新鲜精子(C组)的代谢产物谱,平衡精子(E组),冻融精子(F组)。总的来说,确定了2570种呈阳性模式的代谢物和2306种呈阴性模式的代谢物,分别。经过三组的比较分析,374个差异丰富的代谢物(DAM)在C与E,291DAM在Cvs.F,和189个DAM在Evs.F以阳性模式获得;同时,530DAM在C与E,405DAM在C与F,和193个DAM在Evs.F是在负模式下获得的,分别。DAM在各种代谢途径中显著富集,包括31个途径在Cvs.E,C中的25条途径与F,和E中的28条途径F,分别。其中,在三个比较中发现了65个DAM和25个显著富集的途径,这可能与精子特征和功能密切相关。特别是,功能术语,如TCA循环,不饱和脂肪酸的生物合成,鞘脂代谢,甘氨酸,丝氨酸和苏氨酸代谢,α-亚麻酸代谢,和丙酮酸代谢,以及相关的关键代谢物,如神经酰胺,甜菜碱,胆碱,富马酸,L-苹果酸和L-乳酸,集中在。总之,我们的研究描述了山羊精子中代谢物的组成及其由冷冻保存过程引起的变化,为进一步探索代谢影响山羊精子质量和抗冻性的分子机制奠定了基础。此外,与冷冻和解冻过程相比,低温平衡对精子质量的影响可能需要更多的关注。
Semen cryopreservation results in the differential remodeling of the molecules presented in sperm, and these alterations related to reductions in sperm quality and its physiological function have not been fully understood. Given this, this study aimed to investigate the
cryoinjury mechanism of goat sperm by analyzing changes of the metabolic characteristics in sperm during the cryopreservation process. The ultra-high-performance liquid chromatography-quadrupole time-of-flight mass spectrometry (UHPLC-QTOF-MS) technique was performed to explore metabolite profiles of fresh sperm (C group), equilibrated sperm (E group), and frozen-thawed sperm (F group). In total, 2570 metabolites in positive mode and 2306 metabolites in negative mode were identified, respectively. After comparative analyses among these three groups, 374 differentially abundant metabolites (DAMs) in C vs. E, 291 DAMs in C vs. F, and 189 DAMs in E vs. F were obtained in the positive mode; concurrently, 530 DAMs in C vs. E, 405 DAMs in C vs. F, and 193 DAMs in E vs. F were obtained in the negative mode, respectively. The DAMs were significantly enriched in various metabolic pathways, including 31 pathways in C vs. E, 25 pathways in C vs. F, and 28 pathways in E vs. F, respectively. Among them, 65 DAMs and 25 significantly enriched pathways across the three comparisons were discovered, which may be tightly associated with sperm characteristics and function. Particularly, the functional terms such as TCA cycle, biosynthesis of unsaturated fatty acids, sphingolipid metabolism, glycine, serine and threonine metabolism, alpha-linolenic acid metabolism, and pyruvate metabolism, as well as associated pivotal metabolites like ceramide, betaine, choline, fumaric acid, L-malic acid and L-lactic acid, were focused on. In conclusion, our research characterizes the composition of metabolites in goat sperm and their alterations induced by the cryopreservation process, offering a critical foundation for further exploring the molecular mechanisms of metabolism influencing the quality and freezing tolerance of goat sperm. Additionally, the impacts of equilibration at low temperature on sperm quality may need more attentions as compared to the freezing and thawing process.