{"title":"在体外,公马精子在 22 °C 下的衰老与 Ca2+ 和氧化还原平衡的改变有关,可能通过调节新陈代谢来减缓衰老。","authors":"","doi":"10.1016/j.theriogenology.2024.08.021","DOIUrl":null,"url":null,"abstract":"<div><h3>Background</h3><p>Conservation of equine semen in the liquid state is a central procedure in horse breeding and constitutes the basis of associated reproductive technologies. The intense mitochondrial activity of the stallion spermatozoa increases oxidative stress along the storage period, leading to sperm demise within 24–48 h of storage, particularly when maintained at room temperature. Recently, the relationship between metabolism and oxidative stress has been revealed. The study aimed to extend the period of conservation of equine semen, at room temperature through modification of the metabolites present in the media.</p></div><div><h3>Material and methods</h3><p>Processed ejaculates (n = 9) by single-layer colloid centrifugation were split in different aliquots and extended in Tyrode's basal media, or modified Tyrode's consisting of 1 mM glucose, 1 mM glucose 10 mM pyruvate, 40 mM glucose, 40 mM Glucose 10 mM pyruvate, 67 mM glucose and 67 mM glucose 10 mM pyruvate. At time 0h, and after 24 and 96 h of storage, motility was evaluated by CASA, while mitochondrial production of Reactive oxygen species (ROS), and intracellular Ca<sup>2</sup>+ concentrations were determined via flow cytometry using Mitosox Red and Fluo-4 respectively. ROS and Ca<sup>2+</sup> were estimated as Relative Fluorescence Units (RFU) in compensated, arcsin-transformed data in the live sperm population.</p></div><div><h3>Results</h3><p>After 48 h of incubation, motility was greater in all the 10 mM pyruvate-based media, with the poorest result in the 40 mM glucose (41 ± 1.1 %) while the highest motility was yielded in the 40 mM glucose 10 mM pyruvate aliquot (60.3 ± 3.5 %; <em>P</em> < 0.001); after 96 h of storage highest motility values were observed in the 40 mM glucose 10 mM pyruvate media (23.0 ± 6.2 %) while the lowest was observed in the 1 mM glucose media was 9.2 ± 2.0 % <em>(P</em> < 0.05). Mitochondrial ROS was lower in the 40 mM glucose 10 mM pyruvate group compared to the 40 mM glucose (<em>P</em> < 0.01). Over time Ca<sup>2+</sup> increased in all treatment groups compared to time 0h.</p></div><div><h3>Discussion and conclusion</h3><p>Viable spermatozoa may experience oxidative stress and alterations in Ca2+ homeostasis during prolonged storage, however, these effects can be reduced by regulating metabolism. The 40 mM glucose- 10 mM pyruvate group yielded the highest sperm quality parameters.</p></div>","PeriodicalId":23131,"journal":{"name":"Theriogenology","volume":null,"pages":null},"PeriodicalIF":2.4000,"publicationDate":"2024-08-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.sciencedirect.com/science/article/pii/S0093691X2400342X/pdfft?md5=864a3516573d1173e1becf55ace534b9&pid=1-s2.0-S0093691X2400342X-main.pdf","citationCount":"0","resultStr":"{\"title\":\"In vitro, the aging of stallion spermatozoa at 22 °C is linked to alteration in Ca2+ and redox homeostasis and may be slowed by regulating metabolism\",\"authors\":\"\",\"doi\":\"10.1016/j.theriogenology.2024.08.021\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<div><h3>Background</h3><p>Conservation of equine semen in the liquid state is a central procedure in horse breeding and constitutes the basis of associated reproductive technologies. The intense mitochondrial activity of the stallion spermatozoa increases oxidative stress along the storage period, leading to sperm demise within 24–48 h of storage, particularly when maintained at room temperature. Recently, the relationship between metabolism and oxidative stress has been revealed. The study aimed to extend the period of conservation of equine semen, at room temperature through modification of the metabolites present in the media.</p></div><div><h3>Material and methods</h3><p>Processed ejaculates (n = 9) by single-layer colloid centrifugation were split in different aliquots and extended in Tyrode's basal media, or modified Tyrode's consisting of 1 mM glucose, 1 mM glucose 10 mM pyruvate, 40 mM glucose, 40 mM Glucose 10 mM pyruvate, 67 mM glucose and 67 mM glucose 10 mM pyruvate. At time 0h, and after 24 and 96 h of storage, motility was evaluated by CASA, while mitochondrial production of Reactive oxygen species (ROS), and intracellular Ca<sup>2</sup>+ concentrations were determined via flow cytometry using Mitosox Red and Fluo-4 respectively. ROS and Ca<sup>2+</sup> were estimated as Relative Fluorescence Units (RFU) in compensated, arcsin-transformed data in the live sperm population.</p></div><div><h3>Results</h3><p>After 48 h of incubation, motility was greater in all the 10 mM pyruvate-based media, with the poorest result in the 40 mM glucose (41 ± 1.1 %) while the highest motility was yielded in the 40 mM glucose 10 mM pyruvate aliquot (60.3 ± 3.5 %; <em>P</em> < 0.001); after 96 h of storage highest motility values were observed in the 40 mM glucose 10 mM pyruvate media (23.0 ± 6.2 %) while the lowest was observed in the 1 mM glucose media was 9.2 ± 2.0 % <em>(P</em> < 0.05). Mitochondrial ROS was lower in the 40 mM glucose 10 mM pyruvate group compared to the 40 mM glucose (<em>P</em> < 0.01). Over time Ca<sup>2+</sup> increased in all treatment groups compared to time 0h.</p></div><div><h3>Discussion and conclusion</h3><p>Viable spermatozoa may experience oxidative stress and alterations in Ca2+ homeostasis during prolonged storage, however, these effects can be reduced by regulating metabolism. 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引用次数: 0
摘要
背景马精液的液态保存是马匹繁育的核心程序,也是相关生殖技术的基础。种马精子线粒体的强烈活动增加了贮存期间的氧化应激,导致精子在贮存 24-48 小时内死亡,尤其是在室温下贮存时。最近,新陈代谢与氧化应激之间的关系得到了揭示。本研究旨在通过改变介质中的代谢物来延长马精液在室温下的保存期。材料和方法通过单层胶体离心法处理的精液(n = 9)被分成不同的等分,并在泰罗德基础培养基或由 1 mM 葡萄糖、1 mM 葡萄糖 10 mM 丙酮酸、40 mM 葡萄糖、40 mM 葡萄糖 10 mM 丙酮酸、67 mM 葡萄糖和 67 mM 葡萄糖 10 mM 丙酮酸组成的改良泰罗德培养基中延长保存时间。在储存 0 小时、24 小时和 96 小时后,用 CASA 评估运动能力,同时用 Mitosox Red 和 Fluo-4 通过流式细胞仪分别测定线粒体产生的活性氧(ROS)和细胞内 Ca2+ 浓度。结果培养 48 小时后,在所有 10 mM 丙酮酸培养基中,精子的运动能力都较强,在 40 mM 葡萄糖培养基中,精子的运动能力最弱(41 ± 1.1 %),而 40 mM 葡萄糖 10 mM 丙酮酸等分液的运动能力最高(60.3 ± 3.5 %;P <;0.001);储存 96 小时后,40 mM 葡萄糖 10 mM 丙酮酸培养基的运动能力值最高(23.0 ± 6.2 %),而 1 mM 葡萄糖培养基的运动能力值最低(9.2 ± 2.0 %)(P <;0.05)。与 40 mM 葡萄糖相比,40 mM 葡萄糖 10 mM 丙酮酸组的线粒体 ROS 更低(P <0.01)。讨论和结论有活力的精子在长期储存期间可能会经历氧化应激和 Ca2+ 平衡的改变,但是这些影响可以通过调节新陈代谢来减少。40 mM 葡萄糖-10 mM 丙酮酸组的精子质量参数最高。
In vitro, the aging of stallion spermatozoa at 22 °C is linked to alteration in Ca2+ and redox homeostasis and may be slowed by regulating metabolism
Background
Conservation of equine semen in the liquid state is a central procedure in horse breeding and constitutes the basis of associated reproductive technologies. The intense mitochondrial activity of the stallion spermatozoa increases oxidative stress along the storage period, leading to sperm demise within 24–48 h of storage, particularly when maintained at room temperature. Recently, the relationship between metabolism and oxidative stress has been revealed. The study aimed to extend the period of conservation of equine semen, at room temperature through modification of the metabolites present in the media.
Material and methods
Processed ejaculates (n = 9) by single-layer colloid centrifugation were split in different aliquots and extended in Tyrode's basal media, or modified Tyrode's consisting of 1 mM glucose, 1 mM glucose 10 mM pyruvate, 40 mM glucose, 40 mM Glucose 10 mM pyruvate, 67 mM glucose and 67 mM glucose 10 mM pyruvate. At time 0h, and after 24 and 96 h of storage, motility was evaluated by CASA, while mitochondrial production of Reactive oxygen species (ROS), and intracellular Ca2+ concentrations were determined via flow cytometry using Mitosox Red and Fluo-4 respectively. ROS and Ca2+ were estimated as Relative Fluorescence Units (RFU) in compensated, arcsin-transformed data in the live sperm population.
Results
After 48 h of incubation, motility was greater in all the 10 mM pyruvate-based media, with the poorest result in the 40 mM glucose (41 ± 1.1 %) while the highest motility was yielded in the 40 mM glucose 10 mM pyruvate aliquot (60.3 ± 3.5 %; P < 0.001); after 96 h of storage highest motility values were observed in the 40 mM glucose 10 mM pyruvate media (23.0 ± 6.2 %) while the lowest was observed in the 1 mM glucose media was 9.2 ± 2.0 % (P < 0.05). Mitochondrial ROS was lower in the 40 mM glucose 10 mM pyruvate group compared to the 40 mM glucose (P < 0.01). Over time Ca2+ increased in all treatment groups compared to time 0h.
Discussion and conclusion
Viable spermatozoa may experience oxidative stress and alterations in Ca2+ homeostasis during prolonged storage, however, these effects can be reduced by regulating metabolism. The 40 mM glucose- 10 mM pyruvate group yielded the highest sperm quality parameters.
期刊介绍:
Theriogenology provides an international forum for researchers, clinicians, and industry professionals in animal reproductive biology. This acclaimed journal publishes articles on a wide range of topics in reproductive and developmental biology, of domestic mammal, avian, and aquatic species as well as wild species which are the object of veterinary care in research or conservation programs.