Yildiz C, Kaya A, Aksoy M, Tekeli T
Department of Reproduction and Artificial Insemination, Faculty of Veterinary Medicine, Selçuk University, Konya, Turkey.
Theriogenology. 2000 Sep 1;54(4):579-85. doi: 10.1016/S0093-691X(00)00373-3.
Influence of different sugars supplemented to the extender on the motility, viability and intact acrosome rates of dog spermatozoa during dilution, equilibration and freezing was studied. The ejaculate was divided into 10 aliquots, which were diluted 1:3 with TRIS-citric acid extender containing 240 mMTRIS, 63 mM citric acid, 8% (v/v) glycerol, 20% (v/v) egg yolk and 70 mM sugar, which was either fructose, galactose, glucose, xylose (monosaccharide), lactose, trehalose, maltose, sucrose (disaccharide) or raffinose (trisaccharide). No sugar was added to the extender in the control group. Extended semen samples were cooled to 5 degrees C over 45 min, packaged in 0.25-mL straws, equilibrated for 2 h at 5 degrees C and frozen in liquid nitrogen vapor. Samples were thawed by placing straws into 37 degrees C water for 30 sec. Motility, viable sperm and intact acrosome rates decreased gradually in all groups after equilibration and consecutively freezing (P<0.001). The type of sugar significantly effected motility, viability and acrosomal integrity during equilibration and freezing (P<0.05). Galactose, lactose, trehalose, maltose and sucrose reduced damaged acrosome percentages in equilibrated samples (P<0.05). Sugar supplementation did not enhance motility and viability during equilibration. The disaccharides, except lactose, reduced post-thaw dead sperm and/or damaged acrosome percentages without promoting post-thaw motility (P<0.01), whereas monosaccharides, especially fructose and xylose, improved motility (P<0.05) along with viability and intact acrosome rates (P<0.05). Trehalose, xylose and fructose significantly increased total active sperm rates (motility x live sperm rate x normal acrosome rate) compared to other sugars (P<0.01) and control (P<0.0001) in frozen thawed samples. Therefore, sugar supplementation of the extender influenced post-equilibration and post-thaw sperm quality, and the type or locality of protective impact of the sugar on dog spermatozoa vary according to type of the sugar.
研究了在稀释、平衡和冷冻过程中,向稀释液中添加不同糖类对犬精子活力、存活率和顶体完整率的影响。将射精精液分成10等份,用含有240 mM三羟甲基氨基甲烷(TRIS)、63 mM柠檬酸、8%(v/v)甘油、20%(v/v)蛋黄和70 mM糖类的TRIS - 柠檬酸稀释液按1:3进行稀释,糖类分别为果糖、半乳糖、葡萄糖、木糖(单糖)、乳糖、海藻糖、麦芽糖、蔗糖(二糖)或棉子糖(三糖)。对照组的稀释液中不添加糖类。将稀释后的精液样本在45分钟内冷却至5℃,装入0.25 mL细管中,在5℃平衡2小时,然后在液氮蒸汽中冷冻。将细管置于37℃水中30秒进行解冻。平衡和连续冷冻后,所有组的精子活力、存活精子和顶体完整率均逐渐下降(P<0.001)。糖类类型在平衡和冷冻过程中对精子活力、存活率和顶体完整性有显著影响(P<0.05)。半乳糖、乳糖、海藻糖、麦芽糖和蔗糖降低了平衡样本中顶体受损的百分比(P<0.05)。添加糖类在平衡过程中并未提高精子活力和存活率。除乳糖外的二糖降低了解冻后死精子和/或顶体受损的百分比,但未提高解冻后精子活力(P<0.01),而单糖,尤其是果糖和木糖,提高了精子活力(P<0.05)以及存活率和顶体完整率(P<0.05)。与其他糖类(P<0.01)和对照组(P<0.0001)相比,海藻糖、木糖和果糖在冻融样本中显著提高了总活动精子率(活力×活精子率×正常顶体率)。因此,向稀释液中添加糖类会影响平衡后和解冻后的精子质量,糖类对犬精子的保护作用类型或部位因糖类类型而异。