Beccaglia M, Anastasi P, Chigioni S, Luvoni G C
Department of Veterinary Clinical Sciences, Obstetrics and Gynaecology, University of Milan, Milan, Italy.
Reprod Domest Anim. 2009 Jul;44 Suppl 2:345-9. doi: 10.1111/j.1439-0531.2009.01410.x.
The aims were to evaluate the suitability of a non-commercial TRIS-lecithin (LC) extender and the effect of different concentrations of catalase (CAT) on motility, capacitation status (Chlortetracycline-assay) and zona pellucida (ZP) binding capacity of canine spermatozoa stored at +5 degrees C for 4 days. The sperm-rich fractions of the ejaculates of four stud dogs were divided into four aliquots. After centrifugation, sperm pellets were diluted (200 x 10(6) sperm/ml) in TRIS buffer, citric acid, fructose, antibiotics, supplemented with 20% egg yolk (TRIS-EY) or 0.04% soybean lecithin (TRIS-LC) with CAT (150 or 450 UI/ml) or without CAT, and then preserved at 5 degrees C for 4 days. The results showed that LC is a valid alternative to EY for chilling canine semen, as similar rates of motility, number of uncapacitated spermatozoa and of spermatozoa binding the oocyte ZP were obtained in semen chilled in TRIS-LC or TRIS-EY. Different concentrations of CAT in a TRIS-LC based extender did not improve the quality of semen after chilling. However, a concentration of 150 UI/ml CAT resulted in an increased number of spermatozoa bound to the oocyte ZP, after 4 days of chilling when compared to semen chilled with TRIS-EY and TRIS-LC. In conclusion, an animal protein-free extender with soybean LC, as a replacement of EY, is suitable for 4 days chilling of canine spermatozoa, but the addition of CAT does not improve general semen quality except for a slight effect on sperm-ZP binding.
本研究旨在评估一种非商业性的TRIS-卵磷脂(LC)稀释液的适用性,以及不同浓度的过氧化氢酶(CAT)对犬精子在5℃下储存4天的活力、获能状态(金霉素检测法)和透明带(ZP)结合能力的影响。将四只种公犬射精的富含精子部分分为四个等分试样。离心后,精子沉淀用含有柠檬酸、果糖、抗生素的TRIS缓冲液稀释(200×10⁶精子/ml),添加20%蛋黄(TRIS-EY)或0.04%大豆卵磷脂(TRIS-LC),并添加CAT(150或450 UI/ml)或不添加CAT,然后在5℃下保存4天。结果表明,对于犬精液冷冻保存,LC是EY的有效替代品,因为在TRIS-LC或TRIS-EY中冷冻的精液,其活力、未获能精子数量和与卵母细胞ZP结合的精子数量相似。基于TRIS-LC的稀释液中不同浓度的CAT并不能改善冷冻后精液的质量。然而,与用TRIS-EY和TRIS-LC冷冻的精液相比,浓度为150 UI/ml的CAT在冷冻4天后导致与卵母细胞ZP结合的精子数量增加。总之,用大豆LC替代EY的无动物蛋白稀释液适用于犬精子4天的冷冻保存,但添加CAT除了对精子-ZP结合有轻微影响外,并不能改善精液的总体质量。