Effect of equilibration time on the motility and functional integrity of canine spermatozoa frozen in three different extenders
Effect of equilibration time on the motility and functional integrity of canine spermatozoa frozen in three different extenders
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DOI：
10.1016/j.rvsc.2016.03.010
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发表时间：
2016-06-01
影响因子：
2.4
通讯作者：
Bencharif, D.
中科院分区：
文献类型：
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作者：
Belala, R.;Briand-Amirat, L.;Bencharif, D.
用6%低密度脂蛋白(LDL)、6%脂蛋白(Lipoo)和40%卵黄血浆(EYP)三种冷冻稀释液，研究了冷冻时间对犬精子解冻后活力参数的影响。第二个实验的目的是评估冷冻在三种稀释液中的犬精子在实验一中找到的最佳平衡时间的功能完整性。在第一个实验中，收集7只狗的20个精液，在4个平衡时间(30min、1h、3h和6h)后，将其冷冻在三种稀释剂(低密度脂蛋白、脂蛋白和EYP)中。使用图像分析仪(HT-IVOS 14.0)对解冻后的精液进行评估。平衡时间为6h时，三种延长器的运动和递进运动效果较好。(低密度脂蛋白：58.9%对42.7%；脂蛋白：54.4%对31.9%；EYP：55.4%对40.5%，运动6h对40.5%)。在第二个实验中，6只狗的10个精液在与前一个实验相同的条件下冷冻，平衡时间为6h。在稀释后(TO)、平衡后和解冻后三个不同阶段，对精子膜完整性参数(低渗肿胀试验和SYBR14/碘化丙啶染色)、顶体(FITC-PIZS SAGININ染色)和DNA(吖啶橙染色)进行了评价。解冻后的结果如下：膜完整性(宿主：62；6%vs 58%vs 64.4%；SYBR14/IP：63.6%vs 57.9%vs 64.8%)；顶体完整性(FITC-PSA：79.4%vs 74%vs 76.2%)；DNA完整性(丫啶橙：98.9%vs 98.5%vs 98.7%)。不同稀释液之间无显著差异，因此在不改变精子活力和完整性参数的情况下，6%Lipo6%和40%EYP可替代6%低密度脂蛋白用于犬精子的冷冻-解冻保护。(C)2016爱思唯尔有限公司。保留所有权利。
The present work aimed to assess the effect of equilibration time on post-thaw motility parameters of canine sperm frozen in three extenders: 6% low-density lipoproteins (LDL), 6% liposomes (LIPO), and 40% egg yolk plasma (EYP). A second experiment is aimed at evaluating the functional integrity of canine spermatozoa frozen in the three extenders at the best equilibration time found in the experiment one. In the first experiment, 20 ejaculates harvested from 7 dogs, were frozen in three extenders (LDL, LIPO, and EYP) after four equilibration times (30 min, 1 h, 3 h, and 6 h). The semen was evaluated after thawing using an image analyser (HT-IVOS 14.0). The 6 h equilibration time gave better results of motility and progressive motility in the three studied extenders. (LDL: 58.9% vs. 42.7%; LIPO: 54.4% vs. 31.9%; EYP: 55.4% vs 40.5% for motility 6 vs. 1 h). In the second experiment, 10 ejaculates taken from 6 dogs were frozen under the same conditions as the previous experiment, after 6 h equilibration time. The integrity parameters of the spermatozoal membrane (hypo-osmotic swelling test, and SYBR14/propidium Iodide staining), acrosome (FITC-Pisium sativum Aglutinin staining), and DNA (acridine orange staining) were evaluated at three different stages: post-dilution (TO), post-equilibration, and post-thawing. Post-thaw results were as follows: membrane integrity (HOSt: 62;6% vs 58% vs 64.4%; SYBR14/IP: 63.6% vs 57.9% vs 64.8%); acrosome integrity (FITC-PSA: 79.4% vs 74% vs 76.2%) and DNA integrity (Acridine-orange: 98.9% vs 98.5% vs 98.7%) respectively for LDL vs. LIPO vs. EYP. No significant difference existed between the extenders tested; thus 6%LIPO and 40%EYP could be good candidates for replacement of 6%LDL in the protection of canine sperm during the freeze-thaw process without altering motility and integrity parameters. (C) 2016 Elsevier Ltd. All rights reserved.