L-carnitine supplementation in conventional slow and ultra-rapid freezing media improves motility, membrane integrity, and fertilizing ability of dog epididymal sperm

IF 2.2 2区农林科学 Q1 AGRICULTURE, DAIRY & ANIMAL SCIENCE

Animal Reproduction Science Pub Date : 2024-08-24 DOI:10.1016/j.anireprosci.2024.107580

A.E. Ramón-López , J.P. Fernández-Collahuazo , J.X. Samaniego , J.M. Duma , M.S. Méndez , M.E. Soria , L. Galarza-Álvarez , E. Muñoz-León , D.A. Galarza

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Abstract

This study aimed to assess the impact of L-carnitine (LC) supplementation in conventional-slow (CS) and ultra-rapid (UR) freezing media on post-thaw quality and fertilizing ability of dog epididymal spermatozoa. Sperm samples were collected from 60 epididymides obtained from 30 adult orchiectomized dogs via retrograde flushing. Twenty pooled sperm samples were then created (3 epididymal samples/pool). Four treatments were established according to the freezing method (CS and UR) and LC supplementation (5 and 0 mM [control, Co]): CS-LC5, CS-Co, UR-LC5, and UR-Co. The CS freezing involved exposing 0.25 mL straw to liquid nitrogen vapors (LN₂), while UR freezing submerged 30-µL drops of sperm samples directly into LN₂. Sperm kinematics, membrane integrity, and fertilizing ability (by heterologous in vitro fertilization using bovine oocytes) were evaluated for all treatments. Post-thaw results revealed that the CS freezing treatments resulted in significantly higher values (P < 0.05) of curvilinear and average-path velocities, and beat-cross frequency compared to the UR freezing treatments, regardless of LC supplementation. The CS-LC5 and UR-LC5 treatments cryoprotected the sperm by increasing (P < 0.05) the percentage of ‘live-sperm/intact-acrosome’ compared to their controls treatments CS-Co and UR-Co. Regarding fertilizing ability, the CS-LC5 treatment yielded a higher percentage (P < 0.05) of pronuclei formation compared to both UR treatments. The UR-LC5 treatment, however, obtained greater percentage (P < 0.05) than their control UR-Co. In conclusion, supplementation with L-carnitine in conventional-slow and ultra-rapid freezing improved sperm motility, plasma, and acrosome membranes integrity and fertilizing ability of dog epididymal spermatozoa.

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在常规慢速和超快速冷冻培养基中补充左旋肉碱可提高狗附睾精子的活力、膜完整性和受精能力

本研究旨在评估在常规慢速（CS）和超快速（UR）冷冻培养基中补充左旋肉碱（LC）对狗附睾精子解冻后质量和受精能力的影响。通过逆行冲洗从 30 只睾丸切除的成年狗的 60 个附睾中收集精子样本。然后制作了 20 份精子样本池（每池 3 份附睾样本）。根据冷冻方法（CS 和 UR）和 LC 补充量（5 毫摩尔和 0 毫摩尔[对照组，Co]）确定了四种处理方法：CS-LC5、CS-Co、UR-LC5 和 UR-Co。CS 冻结法是将 0.25 毫升吸管置于液氮蒸汽（LN2）中，而 UR 冻结法是将 30 微升的精子样品直接滴入 LN2 中。对所有处理方法的精子运动学、膜完整性和受精能力（使用牛卵母细胞进行异源体外受精）进行了评估。解冻后的结果显示，与UR冷冻处理相比，CS冷冻处理的曲线速度、平均路径速度和搏动交叉频率的值明显更高（P <0.05），与补充LC的情况无关。与对照组CS-Co和UR-Co相比，CS-LC5和UR-LC5处理的 "活精子/非顶体 "百分比增加（P <0.05），从而对精子起到了冷冻保护作用。在受精能力方面，与两种 UR 处理相比，CS-LC5 处理产生的前核形成比例更高（P < 0.05）。然而，UR-LC5处理比其对照UR-Co获得的百分比更高（P < 0.05）。总之，在传统慢速冷冻和超快速冷冻中补充左旋肉碱可提高狗附睾精子的活力、浆膜和顶体膜的完整性以及受精能力。

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来源期刊

Animal Reproduction Science 农林科学-奶制品与动物科学

CiteScore

4.50

自引率

9.10%

发文量

136

审稿时长

54 days

期刊介绍： Animal Reproduction Science publishes results from studies relating to reproduction and fertility in animals. This includes both fundamental research and applied studies, including management practices that increase our understanding of the biology and manipulation of reproduction. Manuscripts should go into depth in the mechanisms involved in the research reported, rather than a give a mere description of findings. The focus is on animals that are useful to humans including food- and fibre-producing; companion/recreational; captive; and endangered species including zoo animals, but excluding laboratory animals unless the results of the study provide new information that impacts the basic understanding of the biology or manipulation of reproduction. The journal''s scope includes the study of reproductive physiology and endocrinology, reproductive cycles, natural and artificial control of reproduction, preservation and use of gametes and embryos, pregnancy and parturition, infertility and sterility, diagnostic and therapeutic techniques. The Editorial Board of Animal Reproduction Science has decided not to publish papers in which there is an exclusive examination of the in vitro development of oocytes and embryos; however, there will be consideration of papers that include in vitro studies where the source of the oocytes and/or development of the embryos beyond the blastocyst stage is part of the experimental design.