猪精子的体外老化：精子接近度和精液的作用。

In vitro aging of boar spermatozoa: role of sperm proximity and seminal plasma.

机构信息

Unit for Reproductive Medicine of Clinics/Clinic for Pigs and Small Ruminants, University of Veterinary Medicine Hannover, Hannover, Germany.

出版信息

Andrology. 2019 May;7(3):382-390. doi: 10.1111/andr.12600. Epub 2019 Feb 21.

DOI:10.1111/andr.12600

PMID:30793513

Abstract

BACKGROUND

Knowledge on the effect of seminal plasma on sperm function in extended semen during in vitro storage is lacking.

OBJECTIVES

The aim was to examine the interactive role of sperm concentration and seminal plasma concentration on boar sperm function during in vitro aging.

MATERIAL AND METHODS

Experiment 1: Twenty-one boar ejaculates were aliquoted with Beltsville Thawing Solution into five semen doses containing between 32.5 and 8.5 × 10 sperm/mL. Experiment 2: Semen samples (n = 8) containing 18 × 10 or 10 × 10 sperm/mL with their natural amount of seminal plasma and 10 × 10 sperm/mL substituted with autologous seminal plasma to the same concentration as in doses with 18 × 10 sperm/mL were prepared. Experiment 3: Four variants of semen doses containing 18 × 10 or 10 × 10 sperm/mL with either 10% or 0.5% (v/v) seminal plasma were used. Lipid peroxidation was assessed using Bodipy 581/591 in samples (n = 8) with two different sperm concentrations. Semen was examined during 144-h storage at 17 °C by computer-assisted semen analysis and flow cytometry.

RESULTS

Experiment 1: 3D kinematic patterns revealed a concentration- and time-dependent loss of sperm kinematics in samples with < 23 × 10 sperm/mL (p < 0.05). Percent viable spermatozoa with high mitochondria membrane potential were lower (p < 0.05) in samples with < 15 × 10 sperm/mL. Experiment 2: Seminal plasma supplementation in samples with 10 × 10 sperm/mL did not restore the loss of sperm kinematics (p > 0.05). Experiment 3: At 144 h, motility was lowest in samples containing 10 × 10 sperm/mL and 10% (v/v) seminal plasma (p < 0.05). Sperm lipid peroxidation did not differ between samples with different sperm concentration.

CONCLUSION

Long-term exposure to seminal plasma has a negative impact on in vitro-aged boar spermatozoa. Reduced sperm-to-sperm proximity but not the reduction of seminal plasma limits sperm function in long-term stored boar semen.

摘要

背景

关于在体外储存过程中延长精液中精子功能的精液浆的影响，目前知识还很缺乏。

目的

本研究旨在检测精子浓度和精液浆浓度在猪精子体外老化过程中的相互作用。

材料和方法

实验 1：将 21 份猪精液用 Beltsville 解冻液等分，制成 5 份精液剂量，每份精液剂量中精子浓度分别为 32.5 至 8.5×10 个/ml。实验 2：准备 8 份精液样本（分别含有 18×10 或 10×10 个/ml 的精子和其天然量的精液浆，以及将 10×10 个/ml 的精子用自体精液浆替代至与含有 18×10 个/ml 精子的剂量相同的浓度）。实验 3：使用含有 18×10 或 10×10 个/ml 精子且精液浆浓度分别为 10%或 0.5%（v/v）的 4 种精液剂量变体。使用 Bodipy 581/591 在 2 种不同精子浓度的样本中评估脂质过氧化。在 17℃下储存 144 小时期间，通过计算机辅助精液分析和流式细胞术对精液进行检查。

结果

实验 1：3D 运动学模式显示，在<23×10 个/ml 的样本中，精子运动学呈浓度和时间依赖性丧失（p<0.05）。高线粒体膜电位的活精子百分比在<15×10 个/ml 的样本中较低（p<0.05）。实验 2：在 10×10 个/ml 的样本中补充精液浆并没有恢复精子运动学的丧失（p>0.05）。实验 3：在 144 小时时，含有 10×10 个/ml 精子和 10%（v/v）精液浆的样本中的活力最低（p<0.05）。不同精子浓度的样本之间的精子脂质过氧化没有差异。