Yunnan Animal Science and Veterinary Institute, Jindian, Panlong District, Kunming, 650224, China.
Yunnan Provincial Engineering Research Center of Livestock Genetic Resource Conservation and Germplasm Enhancement, Jindian, Panlong District, Kunming, 650224, China.
Sci Rep. 2024 Nov 27;14(1):29489. doi: 10.1038/s41598-024-80556-2.
Results of artificial insemination (AI) are affected by changes in sperm quality and the function throughout collection and preservation procedures. Proteome and metabolome alterations of sperm treated with the different procedures in goat, however, aren't fully understood. To this end, we sought to investigate the impacts of rectal probe electrostimulation (EE) and artificial vagina (AV) semen collection methods on the quality and the cryotolerance of goat sperm, with additional focus on proteomic and metabolomic analyses. Semen samples were collected from Yunshang black goats and categorized into four groups: fresh sperm collected via AV (XAZ), fresh sperm collected via EE (XEZ), frozen sperm post-AV collection (DAZ) and frozen sperm post-EE collection (DEZ). Four comparisons (XAZ vs. XEZ, DAZ vs. XAZ, DEZ vs. XEZ, DAZ vs. DEZ) were performed, respectively. This study first evaluated sperm motility, acrosome integrity, plasma membrane integrity, mitochondrial activity, and reactive oxygen species (ROS) levels. The results indicated that there were no significant differences in fresh sperm quality parameters between the EE and AV methods. However, notable differences emerged post-cryopreservation. Specifically, the AV method proved more advantageous in preserving the motility, integrities of acrosome and plasma membrane, mitochondrial activity of frozen sperm compared to the EE method. Through the multi-omics approaches, a total of 210 differentially abundant proteins (DAPs) related to sperm characteristics and function were identified across the four comparations. Moreover, 32 differentially abundant metabolites (DAMs) were detected. Comprehensive bioinformatics analysis underscored significant molecular pathways in the co-enrichment of DAPs and DAMs, particularly focusing on the citrate cycle, ROS, oxidative phosphorylation, and glycine, serine, and threonine metabolism etc. We elucidated the differential impacts of AV and EE collection methods on the quality and cryotolerance of goat semen from omics perspectives, which offer a critical foundation for further exploration into optimizing semen collection and cryopreservation techniques in goat breeding program.
人工授精（AI）的结果受精子质量和收集及保存过程中功能变化的影响。然而，关于不同处理方法对山羊精子的蛋白质组和代谢组改变还不完全了解。为此，我们试图研究直肠探针电刺激（EE）和人工阴道（AV）精液收集方法对山羊精子质量和冷冻耐受性的影响，并特别关注蛋白质组和代谢组分析。精液样本取自云尚黑山羊，分为四组：通过 AV 收集的新鲜精子（XAZ）、通过 EE 收集的新鲜精子（XEZ）、AV 收集后冷冻的精子（DAZ）和 EE 收集后冷冻的精子（DEZ）。分别进行了四个比较（XAZ 与 XEZ、DAZ 与 XAZ、DEZ 与 XEZ、DAZ 与 DEZ）。本研究首先评估了精子的运动能力、顶体完整性、质膜完整性、线粒体活性和活性氧（ROS）水平。结果表明，EE 和 AV 方法对新鲜精子质量参数没有显著差异。然而，冷冻保存后出现了显著差异。具体而言，与 EE 方法相比，AV 方法在保存冷冻精子的活力、顶体和质膜的完整性、线粒体活性方面更具优势。通过多组学方法，在这四个比较中总共鉴定出了 210 个与精子特征和功能相关的差异丰度蛋白（DAPs）。此外，还检测到 32 个差异丰度代谢物（DAMs）。综合生物信息学分析强调了 DAPs 和 DAMs 共富集的显著分子途径，特别是关注柠檬酸循环、ROS、氧化磷酸化和甘氨酸、丝氨酸和苏氨酸代谢等。我们从组学角度阐明了 AV 和 EE 收集方法对山羊精液质量和冷冻耐受性的不同影响，为进一步探索优化山羊繁殖计划中的精液收集和冷冻保存技术提供了重要基础。
