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巴塔哥尼亚鳚(Eleginops maclovinus)精子在4℃的科特兰培养基中储存后的质量

Patagonian blenny (Eleginops maclovinus) spermatozoa quality after storage at 4 ºC in Cortland medium.

作者信息

Ulloa-Rodríguez Patricio, Contreras Pablo, Dumorné Kelly, Lee-Estevez Manuel, Díaz Rommy, Figueroa Elías, Valdebenito Iván, Risopatrón Jennie, Farías Jorge G

机构信息

Department of Chemistry Engineering, La Frontera University, Temuco, Chile; Center of Biotechnology in Reproduction (CEBIOR-BIOREN), La Frontera University, Temuco, Chile.

Center of Biotechnology in Reproduction (CEBIOR-BIOREN), La Frontera University, Temuco, Chile; School of Aquaculture, Catholic University of Temuco, Temuco, Chile.

出版信息

Anim Reprod Sci. 2018 Oct;197:117-125. doi: 10.1016/j.anireprosci.2018.08.019. Epub 2018 Aug 18.

Abstract

Patagonian blenny (E. maclovinus) is a marine species recently placed in captivity and which are potentially farmable. Understanding and improving its sperm capacity to withstand short-term storage conditions is a key element of initiating an artificial propagation program for this species. The aim of this study is to evaluate the ultrastructure and quality of E. maclovinus sperm during refrigerated storage. To address this objective, scanning electron microscopy (SEM), cytofluorimetric analysis (membrane integrity; reactive oxygen species generation; mitochondrial membrane potential) and cell respiration/mitochondrial-function analysis (ATP content; oxygen consumption) could be useful for optimizing or improving management for artificial reproduction of this species. Severe damage of plasma membranes was observed by SEM at day 7 and 14 of in vitro storage. Analyses of sperm quality were conducted during the 14-day cold storage period when sperm were in diluted (with Cortland solution) and undiluted conditions. When there were diluted conditions, there was greater preservation of motile capacity (from day-7; P < 0.05), membrane integrity (from day-7; P < 0.05), mitochondrial membrane potential (from day-10; P < 0.05) and ATP stores (from day-3; P < 0.05). Oxygen consumption indicators were 18.6% ±14.7% greater in the undiluted samples from day-3, and 32.1%±2.1% of the total spermatozoa had ample amounts of superoxide anion in both undiluted and diluted semen on day-0. The use of Cortland solution extended the viability of sperm when there were longer storage times. Factors that have a greater effect on the quality of semen during storage are reactive oxygen species generation and ATP depletion. In conclusion, Patagonian blenny spermatozoa can be stored at 4 °C between 7 and 10 days using Cortland solution.

摘要

巴塔哥尼亚鳚(E. maclovinus)是一种最近才被圈养且具有潜在养殖价值的海洋物种。了解并提高其精子在短期储存条件下的耐受能力,是启动该物种人工繁殖计划的关键要素。本研究旨在评估冷藏储存期间巴塔哥尼亚鳚精子的超微结构和质量。为实现这一目标,扫描电子显微镜(SEM)、细胞荧光分析(膜完整性;活性氧生成;线粒体膜电位)以及细胞呼吸/线粒体功能分析(ATP含量;氧气消耗)有助于优化或改进该物种人工繁殖的管理。体外储存第7天和第14天时,通过扫描电子显微镜观察到质膜受到严重损伤。在14天的冷藏期内,对精子处于稀释(用科特兰溶液)和未稀释状态时的质量进行了分析。在稀释条件下,运动能力(从第7天起;P < 0.05)、膜完整性(从第7天起;P < 0.05)、线粒体膜电位(从第10天起;P < 0.05)和ATP储备(从第3天起;P < 0.05)的保存情况更好。未稀释样品在第3天时的氧气消耗指标高出18.6%±14.7%,在第0天,未稀释和稀释精液中均有32.1%±2.1%的精子含有大量超氧阴离子。当储存时间较长时,使用科特兰溶液可延长精子的活力。储存期间对精液质量影响较大的因素是活性氧生成和ATP消耗。总之,使用科特兰溶液时,巴塔哥尼亚鳚精子可在4℃下储存7至10天。

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