Al Ahmad M Z Ali, Chatagnon G, Amirat-Briand L, Moussa M, Tainturier D, Anton M, Fieni F
UPSP DGER, Sanitary Risks and Biotechnology of Reproduction, National Veterinary School of Nantes, Nantes, France.
Reprod Domest Anim. 2008 Aug;43(4):429-36. doi: 10.1111/j.1439-0531.2007.00930.x. Epub 2008 Jan 3.
To improve the results obtained with a reference cryopreservation extender (control extender: Triladyl + 20% (v/v) egg yolk + 6.4% (v/v) glycerol) for freezing caprine semen, glutamine was added to 18 split ejaculates at concentrations of 0, 20, 40, 80 and 120 mM (experiment 1). In experiment 2, glutamine was added to 18 split ejaculates at concentrations of 20, 25, 30, 35 and 40 mM. In the third experiment, the egg yolk was replaced with the low-density lipoprotein (LDL) fraction of egg yolk. The quality of frozen then thawed spermatozoa in each extender was compared using computer-assisted semen analysis. In experiment 1, glutamine at concentrations of 20 mm and 40 mm significantly improved sperm motility compared with the control extender. However, at 120 mM, a significant decrease in motility and velocity was observed. In experiment 2, motility, curvilinear velocity and amplitude of lateral head displacement (ALH) were improved in glutamine at 25 mM compared with the control. In experiment 3, 8% LDL and 25 mM glutamine significantly improved sperm motility, straight line velocity and ALH. In the fourth experiment, the quality of the previously defined freezing extender (Triladyl + 8% (v/v) LDL + 25 mM glutamine + 6.4% (v/v) glycerol) was tested by comparing acrosome, tail membrane, plasma membrane and DNA integrity in 18 split ejaculates of frozen then thawed spermatozoa with spermatozoa that had been frozen then thawed in the control extender, and with spermatozoa from fresh, unfrozen sperm. The percentage of spermatozoa with intact acrosomes and tail membranes was significantly higher with the newly defined extender than that observed with the control extender. There was no significant difference in the percentage of spermatozoa with intact DNA between the frozen and fresh semen.
为了提高使用参考冷冻保存稀释液（对照稀释液：Triladyl + 20%（v/v）蛋黄 + 6.4%（v/v）甘油）冷冻山羊精液所获得的结果，将谷氨酰胺以0、20、40、80和120 mM的浓度添加到18份分割射精精液中（实验1）。在实验2中，将谷氨酰胺以20、25、30、35和40 mM的浓度添加到18份分割射精精液中。在第三个实验中，用蛋黄的低密度脂蛋白（LDL）部分替代蛋黄。使用计算机辅助精液分析比较了每种稀释液中冷冻后解冻精子的质量。在实验1中，与对照稀释液相比，20 mM和40 mM浓度的谷氨酰胺显著提高了精子活力。然而，在120 mM时，观察到活力和速度显著下降。在实验2中，与对照相比，25 mM谷氨酰胺提高了精子活力、曲线速度和头部侧向位移幅度（ALH）。在实验3中，8%LDL和25 mM谷氨酰胺显著提高了精子活力、直线速度和ALH。在第四个实验中，通过比较18份分割射精精液中冷冻后解冻精子的顶体、尾膜、质膜和DNA完整性，测试了先前定义的冷冻稀释液（Triladyl + 8%（v/v）LDL + 25 mM谷氨酰胺 + 6.4%（v/v）甘油）的质量，这些精液分别在该稀释液中冷冻后解冻、在对照稀释液中冷冻后解冻以及来自新鲜未冷冻的精子。新定义的稀释液中顶体和尾膜完整的精子百分比显著高于对照稀释液。冷冻精液和新鲜精液中DNA完整的精子百分比没有显著差异。
