Analysis of stallion spermatozoa metabolism using Agilent Seahorse XFp Technology

IF 2.2 2区农林科学 Q1 AGRICULTURE, DAIRY & ANIMAL SCIENCE

Animal Reproduction Science Pub Date : 2024-11-03 DOI:10.1016/j.anireprosci.2024.107633

Jose Manuel Ortiz-Rodriguez , Diego Bucci , Laura Tovar-Pascual , Silvia Granata , Marcella Spinaci , Salvatore Nesci

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引用次数: 0

Abstract

Sperm metabolism consists of a sophisticated network of biochemical reactions and varies between species, resulting in different metabolic strategies for ATP production to maintain sperm functionality. ATP can be produced through glycolysis or in the mitochondria by oxidative phosphorylation (OXPHOS). Since OXPHOS is the predominant metabolic pathway in horses spermatozoa, various assessments of mitochondrial activity are used to evaluate fertility, utilizing techniques such as fluorescent probes analysed via microscopy or flow cytometry, and polarographic electrode assays to measure current flow in response to an applied voltage. Though, these methods are limited by low throughput, as they assess mitochondrial activity at a single time point under a specific treatment condition. This study explores, for the first time, the application of the Agilent Seahorse XFp Technology to evaluate metabolism in stallion spermatozoa. This method enables real-time measurement of cellular metabolism across multiple samples or experimental conditions simultaneously. Ejaculates from eight different stallions were collected, and pools were prepared from three of them. Sperm viability and mitochondrial activity were evaluated by fluorescence microscopy, sperm motility by a computer-assisted sperm analysis system, and sperm metabolism was analysed via the Seahorse XFp analyser. Results confirmed a preference for OXPHOS over glycolysis in ATP production in stallion sperm, with mitochondria contributing significantly to total ATP generation. The Seahorse XFp Technology proved effective in evaluating equine sperm bioenergetics, offering insights into metabolic pathways critical for sperm function. In conclusion, this technology grants a new method for high-throughput analysis of sperm metabolism and quality, which could be applied to future reproductive studies in male equine fertility.

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使用安捷伦海马 XFp 技术分析种马精子的新陈代谢。

精子的新陈代谢由复杂的生化反应网络组成，不同物种的新陈代谢策略各不相同，从而产生不同的 ATP 以维持精子的功能。ATP 可通过糖酵解或线粒体中的氧化磷酸化（OXPHOS）产生。由于 OXPHOS 是马精子中最主要的代谢途径，因此线粒体活性的各种评估方法都被用来评估生育能力，这些方法利用的技术包括通过显微镜或流式细胞仪分析的荧光探针，以及测量电流对外加电压响应的极谱电极测定法。不过，这些方法受限于低通量，因为它们是在特定处理条件下评估单一时间点的线粒体活性。本研究首次探索应用安捷伦海马 XFp 技术评估种马精子的新陈代谢。该方法可同时对多个样本或实验条件下的细胞代谢进行实时测量。采集了八种不同种公马的精子，并从其中三种精子中制备了精子池。精子活力和线粒体活性通过荧光显微镜进行评估，精子活力通过计算机辅助精子分析系统进行评估，精子代谢则通过海马 XFp 分析仪进行分析。结果证实，在公马精子的 ATP 生成过程中，OXPHOS 比糖酵解更优先，线粒体对总 ATP 生成的贡献很大。事实证明，海马 XFp 技术能有效评估马精子的生物能，深入了解对精子功能至关重要的代谢途径。总之，该技术为精子代谢和质量的高通量分析提供了一种新方法，可应用于未来雄性马的生殖研究。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Animal Reproduction Science 农林科学-奶制品与动物科学

CiteScore

4.50

自引率

9.10%

发文量

136

审稿时长

54 days

期刊介绍： Animal Reproduction Science publishes results from studies relating to reproduction and fertility in animals. This includes both fundamental research and applied studies, including management practices that increase our understanding of the biology and manipulation of reproduction. Manuscripts should go into depth in the mechanisms involved in the research reported, rather than a give a mere description of findings. The focus is on animals that are useful to humans including food- and fibre-producing; companion/recreational; captive; and endangered species including zoo animals, but excluding laboratory animals unless the results of the study provide new information that impacts the basic understanding of the biology or manipulation of reproduction. The journal''s scope includes the study of reproductive physiology and endocrinology, reproductive cycles, natural and artificial control of reproduction, preservation and use of gametes and embryos, pregnancy and parturition, infertility and sterility, diagnostic and therapeutic techniques. The Editorial Board of Animal Reproduction Science has decided not to publish papers in which there is an exclusive examination of the in vitro development of oocytes and embryos; however, there will be consideration of papers that include in vitro studies where the source of the oocytes and/or development of the embryos beyond the blastocyst stage is part of the experimental design.