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低质量公猪精液剂量的基质过滤对精子质量的影响。

Effects of matrix filtration of low-quality boar semen doses on sperm quality.

作者信息

Bussalleu E, Pinart E, Rivera M M, Briz M, Sancho S, Yeste M, Casas I, Fàbrega A, Rigau T, Rodriguez-Gil J E, Bonet S

机构信息

Biotecnologia de la Reproducció Animal i Humana, Departament de Biologia, Facultat de Ciències, Universitat de Girona, Girona, Catalonia, Spain.

出版信息

Reprod Domest Anim. 2009 Jun;44(3):499-503. doi: 10.1111/j.1439-0531.2008.01221.x. Epub 2009 Jan 23.

Abstract

The aim of this work was to develop a method to enhance the sperm parameters of ejaculates with low sperm quality from Piétrain boars. Seminal doses were filtered through columns of DEAE Sephadex (length 2.5 +/- 0.5 cm), CM Sephadex (length 5 +/- 0.5 cm), glass wool (length 2 +/- 0.5 cm) or glass bead (length 10 +/- 0.5 cm), with an exit flow rate of 1 ml/40 s in all cases. For each male, 10 ml of the sperm cell-rich fraction diluted at 1 : 6 were filtered. Sperm quality was assessed before and after filtration. Sperm morphology, sperm motility and sperm concentration were determined using the computer program sca((R)) 2002 Production, and sperm viability was evaluated by fluorescence multistaining. Osmotic resistance test and hyperosmotic resistance test were used to determine the osmotic resistance of spermatozoa, whereas l-lactate production estimated the metabolic activity. Results showed a decrease of sperm concentration and osmotic resistance of spermatozoa after filtration in the four matrixes. However, an increase in the frequency of viable spermatozoa with intact acrosome after filtration in glass bead columns and an increase of morphologically normal spermatozoa after filtration in Sephadex CM-50, glass wool and glass bead columns were observed. Despite the decrease in the frequency of progressive motile spermatozoa, l-lactate production and mitochondrial sheath integrity maintained constant after filtration. Our findings indicate that column filtration is an effective method to enhance the sperm quality by selecting viable and morphologically normal spermatozoa without altering DNA, plasma membrane, mitochondrial sheath integrity or inducing premature acrosome reaction.

摘要

本研究的目的是开发一种方法,以提高皮特兰公猪精液质量低下的射精精子参数。精液剂量通过DEAE葡聚糖柱(长度2.5±0.5厘米)、CM葡聚糖柱(长度5±0.5厘米)、玻璃棉柱(长度2±0.5厘米)或玻璃珠柱(长度10±0.5厘米)进行过滤,所有情况下出口流速均为1毫升/40秒。对于每头公猪,将10毫升以1:6稀释的富含精子细胞的部分进行过滤。在过滤前后评估精子质量。使用计算机程序sca((R)) 2002 Production测定精子形态、精子活力和精子浓度,并通过荧光多重染色评估精子活力。采用渗透压抗性试验和高渗抗性试验测定精子的渗透压抗性,而l-乳酸产量估计代谢活性。结果显示,在四种基质中过滤后,精子浓度和精子的渗透压抗性均降低。然而,在玻璃珠柱中过滤后,具有完整顶体的活精子频率增加,在葡聚糖CM-50、玻璃棉和玻璃珠柱中过滤后,形态正常的精子增加。尽管进行性运动精子的频率降低,但过滤后l-乳酸产量和线粒体鞘完整性保持不变。我们的研究结果表明,柱过滤是一种通过选择活的和形态正常的精子来提高精子质量的有效方法,而不会改变DNA、质膜、线粒体鞘完整性或诱导过早的顶体反应。

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