Songsasen N, Yu I, Murton S, Paccamonti D L, Eilts B E, Godke R A, Leibo S P
Audubon Center for Research of Endangered Species and Department of Biological Sciences, University of New Orleans, New Orleans, LA, USA.
Cryobiology. 2002 Feb;44(1):79-90. doi: 10.1016/S0011-2240(02)00009-3.
The objective of this study was to determine osmotic tolerance of canine spermatozoa. The study comprised three experiments: (1) spermatozoa suspended either in an egg yolk-citrate (EYC) extender or in Kenney skim milk extender were exposed to NaCl solutions ranging from 290 to 1500 mOsm; (2) spermatozoa suspended in EYC were exposed to 550 to 1500 mOsm solutions of glucose, galactose, or fructose; and (3) spermatozoa suspended in EYC or glucose-bovine serum albumin (G-BSA) were exposed to 0.6 M (approximately 900 mOsm) or 1.2 M (approximately 1600 mOsm) solutions of glycerol, ethylene glycol (EG), or dimethyl sulfoxide (Me(2)SO). In all experiments, motility and membrane integrity of spermatozoa were assessed after they were diluted into isotonic medium at 37 degrees C. Exposure of canine spermatozoa to solutions of either NaCl or monosaccharides at osmolalities >500 mOsm caused a significant reduction of motility (P<0.01). Motility of spermatozoa was more affected by osmotic stress than their membrane integrity. Osmotic sensitivity of canine spermatozoa was dependent on the type of extender; spermatozoa suspended in the Kenney extender were more resistant to osmotic stress than those suspended in the EYC extender. Despite their sensitivity to exposure to high concentrations of nonpermeating agents, canine spermatozoa were rather resistant to exposure to glycerol and EG. However, Me(2)SO was toxic to canine spermatozoa; motility was substantially reduced after spermatozoa were exposed to 0.6 M Me(2)SO. The type of extender also affected the sensitivity of canine spermatozoa to Me(2)SO; spermatozoa suspended in the EYC extender were more resistant than those suspended in G-BSA. It was concluded that canine spermatozoa are sensitive to osmotic stress, but are tolerant to shrinking and swelling caused by exposure to permeating cryoprotectants.
本研究的目的是确定犬精子的渗透压耐受性。该研究包括三个实验：（1）将悬浮于蛋黄 - 柠檬酸盐（EYC）稀释液或肯尼脱脂乳稀释液中的精子暴露于渗透压范围为290至1500 mOsm的NaCl溶液中；（2）将悬浮于EYC中的精子暴露于渗透压为550至1500 mOsm的葡萄糖、半乳糖或果糖溶液中；（3）将悬浮于EYC或葡萄糖 - 牛血清白蛋白（G - BSA）中的精子暴露于渗透压为0.6 M（约900 mOsm）或1.2 M（约1600 mOsm）的甘油、乙二醇（EG）或二甲基亚砜（Me₂SO）溶液中。在所有实验中，精子在37℃稀释到等渗培养基后评估其活力和膜完整性。犬精子暴露于渗透压>500 mOsm的NaCl或单糖溶液中会导致活力显著降低（P<0.01）。精子活力比其膜完整性更易受渗透压应激的影响。犬精子的渗透压敏感性取决于稀释液的类型；悬浮于肯尼稀释液中的精子比悬浮于EYC稀释液中的精子对渗透压应激更具抵抗力。尽管犬精子对暴露于高浓度非渗透性试剂敏感，但它们对暴露于甘油和EG具有相当的抵抗力。然而，Me₂SO对犬精子有毒性；精子暴露于0.6 M Me₂SO后活力大幅降低。稀释液的类型也影响犬精子对Me₂SO的敏感性；悬浮于EYC稀释液中的精子比悬浮于G - BSA中的精子更具抵抗力。得出的结论是，犬精子对渗透压应激敏感，但对暴露于渗透性冷冻保护剂引起的收缩和肿胀具有耐受性。
