Changes in Kinetic Parameters and Cytological Characteristics of Rooster Spermatozoa under the Influence of Technological Factors

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Abstract

The problems of fertility reducing of rooster semen in the cycle “native sperm–equilibration–short-term and long-term storage (cryopreservation)” are urgent. The purpose of this study was to determine the effect of different methods of preparation (centrifugation or filtration) of rooster semen on its quality characteristics, depending on the method of removing possible pollutions; to evaluate the change in the composition of the cytosol of spermatozoa of native sperm under the influence of dilution and during short-term storage. Materials and methods. Semen of roosters (n = 22) of the Russian white breed was used. Experiment 1: semen was divided into 3 aliquots: I—diluted with synthetic cryoprotective medium LCM in a ratio of 1:1, II—filtered semen diluted with medium (membrane pore diameter 0.2 μm), III—centrifuged (at 3000 rpm in for 10 minutes). Native and frozen/thawed sperm were evaluated in terms of damage to spermatozoa membranes, chromatin, and acrosomes. The composition of carbohydrates and polyols of native spermatozoa was assessed under the influence of dilution and after storage (3 h). The advantage of filtration as a method of technological preparation of semen compared to centrifugation in terms of progressive motility (with rectilinear-translational movement) of sperm (41.0% versus 27.0%) and chromatin damage (43.4% versus 66.4%) has been shown. The same advantage was observed in frozen/thawed sperm filtered before freezing in terms of progressive motility (25.5% vs. 5.5%) and chromatin damage—16.5% vs. 33.6%, respectively. Semen filtration, as a method of technological processing of rooster semen, can be an effective additional step in the preparation of semen for artificial insemination and/or short-term storage. The main component in the composition of the cytosol of native spermatozoa, according to the content of carbohydrates and polyols, was inositol —73.7% of Σ carbohydrates and polyols. The level of inositol decreased during storage by 6.5 times (from 0.030 to 0.007 mg/mL). The data obtained let us suppose the role of inositol as the main antioxidant in the cytosol of spermatozoa. Technological factors of storing rooster semen in various modes (short-term at a temperature of 5°C and long-term at a temperature of –196°C) have a significant impact on the ratio of sperm cytosol components (carbohydrates and polyols). A significant, 2.5‑fold decrease in the relative content of inositol in the cytosol of frozen/thawed spermatozoa, compared with the indicators of native semen, allows us to recommend the introduction of the antioxidant inositol into the composition of cryoprotective media for rooster semen.

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技术因素影响下公鸡精子动力学参数和细胞学特征的变化
摘要 公鸡精液在 "原生精子-校准-短期和长期贮存(冷冻保存)"周期中降低受精率的问题迫在眉睫。本研究的目的是确定公鸡精液的不同制备方法(离心或过滤)对其质量特性的影响,这取决于去除可能的污染的方法;评估在稀释的影响下和短期储存期间原生精子精子细胞质成分的变化。材料和方法使用俄罗斯白种公鸡(n = 22)的精液。实验 1:精液分为 3 等分:I 用合成低温保护介质 LCM 按 1:1 的比例稀释;II 用介质稀释过滤精液(膜孔直径 0.2 μm);III 离心(3000 转/分,10 分钟)。从精子膜、染色质和顶体的损伤角度对原生精子和冷冻/解冻精子进行了评估。在稀释和储存(3 小时)后,对原生精子的碳水化合物和多元醇成分进行了评估。与离心法相比,过滤法作为精液技术制备方法在精子的渐进运动(直线-横向运动)(41.0% 对 27.0%)和染色质损伤(43.4% 对 66.4%)方面具有优势。在冷冻前过滤的冷冻/解冻精子中也观察到了同样的优势,即精子的渐进运动(25.5% 对 5.5%)和染色质损伤(16.5% 对 33.6%)。精液过滤作为公鸡精液的一种技术处理方法,可以成为人工授精和/或短期储存精液制备过程中的一个有效补充步骤。根据碳水化合物和多元醇的含量,本地精子细胞液组成的主要成分是肌醇--占Σ碳水化合物和多元醇的73.7%。在储存过程中,肌醇的含量下降了 6.5 倍(从 0.030 毫克/毫升降至 0.007 毫克/毫升)。这些数据表明肌醇是精子细胞液中的主要抗氧化剂。以不同模式(5°C 温度下的短期储存和 -196°C 温度下的长期储存)储存公鸡精液的技术因素对精子细胞溶液成分(碳水化合物和多元醇)的比例有显著影响。与原生精液的指标相比,冷冻/解冻精子细胞质中肌醇的相对含量明显下降了 2.5 倍,因此我们建议在公鸡精液低温保护介质的成分中加入抗氧化剂肌醇。
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