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精子冷冻保存影响解冻后的活力,但不影响巴西鱼 Brycon insignis(Characiformes)的胚胎发生或幼虫生长。

Sperm cryopreservation affects postthaw motility, but not embryogenesis or larval growth in the Brazilian fish Brycon insignis (Characiformes).

机构信息

Department of Animal Science, Federal University of Lavras (UFLA), Lavras, Minas Gerais, Brazil.

出版信息

Theriogenology. 2012 Sep 1;78(4):803-10. doi: 10.1016/j.theriogenology.2012.03.028. Epub 2012 Apr 26.

Abstract

Sperm cryopreservation is an important method for preserving genetic information and facilitating artificial reproduction. The objective was to investigate whether the cryopreservation process affects postthaw sperm motility, embryogenesis, and larval growth in the fish Brycon insignis. Sperm was diluted in methyl glycol and Beltsville Thawing solution, frozen in a nitrogen vapor vessel (dry shipper) and stored in liquid nitrogen. Half of the samples were evaluated both subjectively (% of motile sperm and motility quality score-arbitrary grading system from 0 [no movement] to 5 [rapidly swimming sperm]) and in a computer-assisted sperm analyzer (CASA; percentage of motile sperm and velocity). The other half was used for fertilization and the evaluation of embryogenesis (cleavage and gastrula stages), hatching rate, percentage of larvae with normal development and larval growth up to 112 days posthatching (dph). Fresh sperm was analyzed subjectively (percentage of motile sperm and motility quality score) and used as the control. In the subjective analysis, sperm motility significantly decreased from 100% motile sperm and quality score of 5 in fresh sperm to 54% motile sperm and quality score of 3 after thawing. Under computer-assisted sperm analyzer evaluation, postthaw sperm had 67% motile sperm, 122 μm/sec of curvilinear velocity, 87 μm/sec of straight-line velocity and 103 μm/sec of average path velocity. There were no significant differences between progenies (pooled data) for the percentage of viable embryos in cleavage (62%) or gastrula stages (24%) or in the hatching rate (24%), percentage of normal hatched larvae (93%), larval body weight (39.8 g), or standard length (12.7 cm) at 112 days posthatching. Based on these findings, cryopreserved sperm can be used as a tool to restore the population of endangered species, such as B. insignis, as well as for aquaculture purposes, without any concern regarding quality of the offspring.

摘要

精子冷冻保存是保存遗传信息和促进人工繁殖的重要方法。本研究旨在探讨冷冻过程是否会影响鱼 Brycon insignis 解冻后精子的活力、胚胎发生和幼虫生长。精子在甲基乙二醇和贝尔茨维尔解冻液中稀释,在氮蒸气容器(干运器)中冷冻,然后储存在液氮中。一半的样本进行主观评估(运动精子的百分比和运动质量评分-从 0[无运动]到 5[快速游动精子]的任意分级系统)和计算机辅助精子分析(CASA;运动精子的百分比和速度)。另一半用于受精和胚胎发生(卵裂和原肠胚阶段)的评估、孵化率、正常发育幼虫的百分比以及孵化后 112 天(dph)的幼虫生长。新鲜精子进行主观分析(运动精子的百分比和运动质量评分)并作为对照。在主观分析中,精子活力从新鲜精子的 100%运动精子和 5 分的运动质量评分显著下降到解冻后的 54%运动精子和 3 分的运动质量评分。在计算机辅助精子分析评估中,解冻后精子的运动精子比例为 67%,曲线速度为 122μm/sec,直线速度为 87μm/sec,平均路径速度为 103μm/sec。卵裂(62%)或原肠胚(24%)期的胚胎存活率、孵化率(24%)、正常孵化幼虫的百分比(93%)、幼虫体重(39.8g)或标准体长(12.7cm)在 112 天孵化后均无显著差异(合并数据)。基于这些发现,冷冻保存的精子可用于恢复濒危物种(如 B. insignis)的种群,也可用于水产养殖,而不必担心后代的质量。

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