Assessment of sperm quality: a flow cytometric approach
Assessment of sperm quality: a flow cytometric approach
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DOI：
10.1016/s0378-4320(01)00160-9
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发表时间：
2001-12-03
影响因子：
2.2
通讯作者：
Graham, JK
中科院分区：
文献类型：
--
作者：
Graham, JK
多年来，科学家们一直在寻求开发能够准确预测精液样本受精能力的实验室分析方法。然而，由于问题的复杂性，这一目标已被证明是难以实现的，而且很可能很难实现。这个问题的部分原因在于精子必须具备使卵子受精的许多特性，以及实验室分析如何同时评估所有这些特性。精液样本中活动精子的百分比是最常用来评估精液质量的指标。然而，这一分析与精液样本的受精能力没有高度相关。运动测定法不能很好地与生育能力联系起来的一个原因是，我们只评估了精子与卵母细胞受精所必须具备的众多属性中的一个。测量多个精子属性的问题之一是所需的时间和成本。使用流式细胞术，可以同时测量精子细胞中的多种精子属性，包括细胞活力、顶体完整性和线粒体功能。此外，精子的获能能力和顶体反应，以及精子的染色体完整性可以用流式细胞术测量。流式细胞术允许我们在不到1分钟的时间内以合理的成本评估50,000个精子。虽然流式细胞术是评估许多精子属性的有力工具，但它不能评估精子与卵母细胞受精所需的所有属性。因此，也正在开发实验室分析方法来评估精子的能力：(1)结合卵母细胞的能力，通过评估精子在体外与鸡蛋卵黄周膜结合的能力；(2)经膜不稳定化合物处理后，体外发生顶体反应；(3)在体外穿透卵母细胞。当使用来自多个精子分析的数据时，与精液样本的受精潜力具有更高的相关性。例如，在一项利用五匹种马进行的研究中，精液样本中活动精子的百分比与生育能力相关性较差（r(2) = 0.22），然而，当精子活力、活力和进入无带仓鼠卵母细胞的渗透率数据一起使用时，这些数据解释了72%的种马生育能力差异（r = 0.849; [Theriogenology 46(1996) 559]）。有了一系列的测试，可以评估许多不同的精子属性，研究人员应该能够更准确地估计精液样本的受精潜力。(C) 2001年Elsevier Science B.V.出版
For many years, scientists have sought to develop laboratory assays that accurately predict the fertilizing capacity of a semen sample. This goal, however, has proven elusive and will most likely be very difficult to achieve, due to the complex nature of the problem. Part of the problem results from the many attributes that a spermatozoon must possess to fertilize an egg, and how laboratory assays can evaluate all of these attributes simultaneously. The percentage of motile sperm in a sample is most commonly used to evaluate semen quality. This assay, however, is not highly correlated with the fertilizing capacity of semen samples. One reason motion assays do not correlate well with fertility is that we are evaluating only one of many attributes that a sperm must possess to fertilize an oocyte. One of the problems of measuring multiple sperm attributes is the time and cost required. Using flow cytometric assays, multiple sperm attributes, including cell viability, acrosomal integrity, and mitochondrial function, can be measured simultaneously in sperm cells. In addition, the ability of sperm to undergo capacitation and the acrosome reaction, as well as the chromosomal integrity of sperm can be measured using flow cytometry. Flow cytometry permits us to evaluate 50,000 sperm in less then 1 min and at reasonable cost. Although flow cytometry is a powerful tool for evaluating many sperm attributes, it cannot evaluate all of the attributes a sperm cell requires to fertilize an oocyte. Therefore, laboratory assays are also being developed to evaluate the ability of sperm: (1) to bind to the oocyte, by evaluating the ability of sperm to bind to the perivitelline membrane of the hen egg in vitro; (2) to undergo an acrosome reaction in vitro, after treatment with membrane destabilizing compounds; and (3) to penetrate oocytes in vitro. When data from multiple sperm assays are used, higher correlations with the fertilizing potential of a semen sample is achieved. For example, in a study conducted utilizing five stallions, the percentage of motile sperm in semen samples correlated poorly with fertility (r(2) = 0.22), however, when data for sperm motility, viability and penetration rates into zona-free hamster oocytes were utilized together, these data explained 72% of the differences in the fertility of the stallions (r = 0.849; [Theriogenology 46 (1996) 559]). Armed with a battery of tests, which evaluate many different sperm attributes, researchers should be able to more accurately estimate the fertilizing potential of semen samples. (C) 2001 Published by Elsevier Science B.V.