Paul R K, Balaganur K, Bahire S V, Kumar D, Singh R
Division of Animal Physiology and Biochemistry, Indian Council of Agricultural Research - Central Sheep and Wool Research Institute, Avikanagar, via Jaipur, Rajasthan, Pin-304 501, India.
Reprod Fertil Dev. 2018 Oct;30(11):1389-1401. doi: 10.1071/RD18063.
Mammalian spermatozoa remain immotile and metabolically inactive in the cauda epididymidis, thus maintaining fertility for several weeks. The aim of this study was to functionally characterise and evaluate the effect of cauda epididymal plasma (CEP) on liquid preservation of ram spermatozoa. Four experiments were conducted to investigate the effects of: (1) CEP and its fractions on sperm motility; (2) CEP (10%, 15%, 20% v/v) on liquid preservation of ram spermatozoa; (3) seminal plasma (SP; 20%, 30%, 50% v/v) on liquid-preserved spermatozoa; and (4) both CEP and post-storage SP treatment on sperm characteristics. Biochemical characterisation of ram CEP revealed high protein (30.9mgmL-1), catalase (68.9IUmL-1), alkaline phosphatase (17.5IUmL-1) activities and total antioxidant capacity (1112µM Trolox equivalent). Progressive motility of prewashed cauda spermatozoa was reduced (P<0.05) by CEP or its protein-rich fraction compared with protein-free plasma or phosphate-buffered saline. After 48h storage, total motility, rapid motility (average path velocity >75µms-1; 53.9%, 73.5% and 71.8% with 0, 15% and 20% CEP respectively) and straight line velocity (86.3, 102.1 and 102.4µms-1 with 0, 15% and 20% CEP respectively) were significantly (P<0.05) higher in the CEP-treated groups than the control. Viability and acrosomal integrity were similar between groups; however, functional membrane integrity was higher (P<0.05) in the 15% CEP-treated group. Treatment of liquid-preserved spermatozoa with either 20%, 30% or 50% SP improved (P<0.05) rapid motility and kinematics at each time point of storage compared with control. In conclusion, liquid preservation of ram spermatozoa in the presence of 15% or 20% CEP and post-storage treatment with SP significantly improve sperm characteristics.
哺乳动物的精子在附睾尾部保持不活动且代谢不活跃状态，从而维持数周的生育能力。本研究的目的是对附睾尾血浆（CEP）进行功能特性鉴定，并评估其对绵羊精子液体保存的影响。进行了四项实验来研究以下方面的影响：（1）CEP及其组分对精子活力的影响；（2）CEP（10%、15%、20% v/v）对绵羊精子液体保存的影响；（3）精浆（SP；20%、30%、50% v/v）对液体保存精子的影响；以及（4）CEP和储存后SP处理对精子特性的影响。绵羊CEP的生化特性显示其具有高蛋白（30.9mg/mL-1）、过氧化氢酶（68.9IU/mL-1）、碱性磷酸酶（17.5IU/mL-1）活性以及总抗氧化能力（1112µM Trolox当量）。与无蛋白血浆或磷酸盐缓冲盐水相比，CEP或其富含蛋白质的组分可降低（P<0.05）预洗附睾尾精子的前向运动能力。储存48小时后，CEP处理组的总活力、快速运动能力（平均路径速度>75µm/s-1；分别添加0%、15%和20% CEP时为53.9%、73.5%和71.8%）以及直线速度（分别添加0%、15%和20% CEP时为86.3、102.1和102.4µm/s-1）均显著高于对照组（P<0.05）。各组之间的活力和顶体完整性相似；然而，15% CEP处理组的功能膜完整性更高（P<0.05）。与对照组相比，用20%﹑30%或50% SP处理液体保存的精子，在每个储存时间点均能提高（P<0.05）快速运动能力和运动学参数。总之，在15%或20% CEP存在的情况下对绵羊精子进行液体保存，并在储存后用SP处理，可显著改善精子特性。
