PROTECTIVE ENVIRONMENTS FOR ANIMAL SPERM: EVOLUTION OF METHODS AND TOPICAL ASPECTS (REVIEW ARTICLE)

A. Sushko, G. Zhegunov, M. Savelieva, Larysa Yeletska, Irina Martinyk
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Abstract

A retrospective review of domestic and foreign sources of literature is presented, as well as data of published own research on cryopreservation of animal sperm. The main historical stages of the creation of protective environments for deep freezing of sperm are given. In the 30s of the last century, a phenomenon characterized by the death of spermatozoa upon sharp cooling in the range of positive temperatures was discovered. It is called temperature shock of sperm. To prevent it, it is proposed to add substances containing phospholipids to the composition of diluents. Such environments can contain both simple components - native chicken egg yolk or milk, and high-tech - lipoproteins, isolated phospholipids of various origins. To stabilize protein-lipid complexes of plasma membranes and acrosomes of sperm during the cooling process, carbohydrates are added to the diluents. Sugars are components of energy supply for sperm and, along with salts, they are the main osmotic regulators. A combination of two or three carbohydrates in the medium was traditionally considered necessary. However, the Kharkiv school of reproductive specialists has proven the possibility of creating effective protective environments using only one sugar - sucrose or lactose - based on considerable practical experience. The effectiveness of germ cell freezing is shown depending on the cryoprotectants used. Glycerin is the first known endocellular cryoprotectant, which is still unsurpassed in sperm cryopreservation. Our own experimental data on the effect of combinations of glycerol with substances from the amide group on the main biological indicators of sperm after deconservation are presented. Cryoprotectants dimethylacetamide (DMAC) and dimethylformamide (DMF) were tested in own experiments on stallion semen. The experiments studied the effect of different concentrations of the above-mentioned penetrating cryoprotectants both on the main physiological characteristics of stallion sperm (motility, survival), and on the degree of damage to the membrane apparatus of sperm. The effectiveness of certain combinations of these substances has been proven. Methods of preventing the negative impact of oxygen and the development of lipid peroxidation processes in sperm during cryopreservation are presented. The concept of using additional hormonal components in diluents, in particular prostaglandin F2a, is revealed. The materials related to the effect on the quality of reproductive cells of healing preparations are displayed. Keywords: artificial insemination, environments, semen, animals, bulls, stallions, cryoprotectants, freezing.
动物精子保护环境:方法的演变和热点问题(综述文章)
本文回顾了国内外有关动物精子冷冻保存的文献,并对已发表的动物精子冷冻保存研究进行了综述。给出了精子深度冷冻保护环境产生的主要历史阶段。在上世纪30年代,人们发现了一种现象,即在正温度范围内急剧冷却时精子死亡。这被称为精子温度冲击。为了防止它,建议在稀释剂的组成中加入含有磷脂的物质。这样的环境既可以包含简单的成分——天然的鸡蛋蛋黄或牛奶,也可以包含高科技的成分——脂蛋白,即各种来源的分离磷脂。为了在冷却过程中稳定质膜和精子顶体的蛋白脂复合物,在稀释剂中添加了碳水化合物。糖是精子能量供应的组成部分,与盐一起,它们是主要的渗透调节因子。传统上认为,两种或三种碳水化合物的混合物是必要的。然而,哈尔科夫生殖专家学校已经证明,根据相当多的实践经验,只使用一种糖——蔗糖或乳糖——就有可能创造有效的保护环境。生殖细胞冷冻的有效性取决于所使用的冷冻保护剂。甘油是第一种已知的细胞内冷冻保护剂,在精子冷冻保存中仍然是无与伦比的。我们自己的实验数据的影响,甘油与物质从酰胺组对精子保存后的主要生物学指标的影响。对冷冻保护剂二甲基乙酰胺(DMAC)和二甲基甲酰胺(DMF)在种马精液中的应用进行了试验。本实验研究了不同浓度的上述穿透性冷冻保护剂对种马精子主要生理特性(活力、存活)和对精子膜器官损伤程度的影响。这些物质的某些组合的有效性已得到证实。介绍了在冷冻保存期间防止氧的负面影响和精子中脂质过氧化过程的发展的方法。在稀释剂中使用额外激素成分的概念,特别是前列腺素F2a。展示了愈合制剂对生殖细胞质量影响的有关材料。关键词:人工授精,环境,精液,动物,公牛,种马,冷冻保护剂,冷冻
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