The immobilization of spermatozoa by freezing and thawing and the protective action of glycerol.
The immobilization of spermatozoa by freezing and thawing and the protective action of glycerol.
复制标题
冷冻和解冻对精子的固定以及甘油的保护作用。
作者：
By J. E. Lovelock;And;C. Polge
Polge、Smith和Parkes（1949）发现，如果在悬浮液中加入甘油，精子可以冷冻和解冻而不丧失活力。在这一发现之后，家禽（Polge，1951）、公牛（Polge & Rowson，1952）和鲱鱼（Blaxter，1953）的精子在冷冻状态下长期保存而不丧失功能。这种方法的实际应用的重要性已经讨论过（Polge & Parkes，1952）。通过观察到甘油也可保护红细胞免受冷冻和解冻的不利影响，从而促进了甘油保护作用的研究（Smith，1950），然后描述了冷冻和解冻时红细胞发生损伤的性质以及甘油提供的保护作用（Lovelock，1953 a，B）。简单地说，冷冻和解冻引起的溶血是由于当水以冰的形式被除去时，细胞所暴露的浓缩盐溶液的破坏作用。在甘油的存在下，在低于凝固点的温度下的电解质浓度充分降低，以充分解释由甘油提供的保护。由于冻融引起红细胞溶血的过程和甘油的保护作用明显很简单，因此观察其他活细胞在冻融过程中的行为是很有意义的。看来，冷冻效应的这种临时解释最好用对高渗盐溶液具有广泛敏感性的活细胞来检验。使用比溶血更精细的损伤标准似乎也是可取的。不同动物的精子满足这两种要求。兔子的皮肤对高渗盐溶液特别敏感（Emmens，1948），而鲱鱼的皮肤显然能耐受至少和海水一样强的盐溶液。此外，对精子活动力的观察，即使有些缺乏精确度，也为评估冷冻损伤提供了一个不同的敏感标准。本文报道了冷冻和解冻对家兔、公牛、家禽和青鱼精子活力的影响。还描述了甘油对这些效应的修饰。
It was discovered by Polge, Smith & Parkes (1949) that spermatozoa could be frozen and thawed without loss of motility if glycerol was included in their suspending medium. Following this discovery the spermatozoa of the domestic fowl (Polge, 1951), ofthe bull (Polge & Rowson, 1952) and ofthe herring (Blaxter, 1953) have been preserved without loss of function for long periods in the frozen state. The importance of the practical applications of this method has been discussed (Polge & Parkes, 1952). The investigation of the protective action of glycerol was facilitated by the observation that red blood cells also were protected against the adverse effects of freezing and thawing by means of glycerol (Smith, 1950), and the nature of the damage occurring to red blood cells when they are frozen and thawed, and of the protection given by glycerol was then described (Lovelock, 1953a, b). Briefly, haemolysis by freezing and thawing is due to the destructive action of the concentrated salt solution to which the cells are exposedwhen water is removed as ice. In the presence of glycerol the electrolyte concentration at temperatures below the freezing point is sufficiently reduced to explain in full the protection afforded by glycerol. The apparent simplicity of the processes causing the haemolysis of red blood cells by freezing and thawing, and of the protective action of glycerol, made it ofsome interest to observe the behaviour of other living cells on freezing and thawing. It seemed that this provisional explanation of the effects of freezing could best be tested using living cells with a wide range of sensitivity to hypertonic salt solutions. It also seemed desirable to use a criterion of damage more delicate than haemolysis. Both of these requirements are met by the spermatozoa of different animals. Those of the rabbit are particularly sensitive to hypertonic salt solutions (Emmens, 1948), while those of the herring clearly will tolerate salt solutions at least as strong as sea water. In addition, the observation of the motility of spermatozoa, even if somewhat lacking in precision, provides a different and sensitive criterion for the assessment of freezing damage. This paper records the effects of freezing and thawing upon the motility ofthe spermatozoa of the rabbit, the bull, the fowl and the herring. The modification of these effects by glycerol is also described.