Yamashiro Hideaki, Toyomizu Masaaki, Kikusato Motoi, Toyama Natsuki, Sugimura Satoshi, Hoshino Yumi, Abe Hiroyuki, Moisyadi Stefan, Sato Eimei
Laboratory of Animal Reproduction, Graduate School of Agricultural Science, Tohoku University, Sendai, Japan.
J Am Assoc Lab Anim Sci. 2010 Mar;49(2):160-6.
We evaluated the cryosurvival of rat epididymal sperm preserved in raffinose-modified Krebs-Ringer bicarbonate-egg yolk extender supplemented with various energy-yielding substrates (glucose, pyruvate, lactate, and ATP) and assessed the effect on sperm oxygen consumption. The incubation of sperm at 37 degrees C for 10 min in lactate-free extender decreased sperm motility and oxygen consumption before and after thawing compared with those of sperm in glucose- and pyruvate-free mediums. We then focused on the effect of supplementing the extender with lactate (0, 10.79, 21.58, 32.37, and 43.16 mM) and found that sperm frozen and thawed in extender supplemented with 32.37 mM lactate exhibited the highest motility. When we supplemented extender containing 32.37 mM lactate with ATP (0, 0.92, 1.85, 3.70, and 5.55 mM), sperm frozen and thawed in the extender supplemented with 1.85 mM ATP exhibited considerably higher motility and viability than those of sperm frozen and thawed in ATP-free extender. These results provide the first evidence that supplementation of the raffinose-modified Krebs-Ringer bicarbonate-egg yolk extender with 32.37 mM lactate and 1.85 mM ATP increases of number of motile sperm before freezing and enhances the cryosurvival of rat sperm. These supplements to the extender may enhance sperm cryosurvival by improving the metabolic capacity of sperm before freezing.
我们评估了保存在添加了各种能量产生底物（葡萄糖、丙酮酸、乳酸和ATP）的棉子糖修饰的 Krebs-Ringer 碳酸氢盐-蛋黄稀释液中的大鼠附睾精子的冷冻存活率，并评估了其对精子耗氧量的影响。与在不含葡萄糖和丙酮酸的培养基中的精子相比，在不含乳酸的稀释液中于37℃孵育10分钟的精子，在解冻前后的活力和耗氧量均降低。然后，我们重点研究了在稀释液中添加乳酸（0、10.79、21.58、32.37和43.16 mM）的效果，发现用添加了32.37 mM乳酸的稀释液冷冻和解冻的精子具有最高的活力。当我们在含有32.37 mM乳酸的稀释液中添加ATP（0、0.92、1.85、3.70和5.55 mM）时，用添加了1.85 mM ATP的稀释液冷冻和解冻的精子，其活力和存活率明显高于在不含ATP的稀释液中冷冻和解冻的精子。这些结果首次证明，在棉子糖修饰的Krebs-Ringer碳酸氢盐-蛋黄稀释液中添加32.37 mM乳酸和1.85 mM ATP可增加冷冻前活动精子的数量，并提高大鼠精子的冷冻存活率。向稀释液中添加这些物质可能通过提高冷冻前精子的代谢能力来增强精子的冷冻存活率。
