Agca Yuksel, Liu Jun, Mullen Steve, Johnson-Ward Jill, Gould Kenneth, Chan Anthony, Critser John
Comparative Medicine Center, College of Veterinary Medicine, University of Missouri, 1600 East Rollins, Room E-109, Columbia, MO 65211, USA.
J Androl. 2005 Jul-Aug;26(4):470-7. doi: 10.2164/jandrol.04169.
Osmotic properties of chimpanzee spermatozoa were studied at 22 degrees C. An electronic particle counter was used to determine the isosmotic cell volume, and the volume response after exposure to four commonly used cryoprotectants: dimethyl sulfoxide, glycerol, propylene glycol, and ethylene glycol. The data were analyzed to determine the hydraulic conductivity and the permeability coefficients for the four cryoprotectants. The osmotically inactive volume fraction was determined using a Boyle van't Hoff plot of cells exposed to sodium chloride solutions. A computer-assisted semen analysis system was used to determine the osmotic tolerance of chimp spermatozoa, as well as the effects of a one-step addition and removal of 1 M permeating cryoprotectant on sperm motility. The isosmotic volume of chimpanzee sperm is 27.7 microm3. The osmotically inactive cell fraction is 69%. Hydraulic conductivity was higher in the presence of ethylene glycol: 4.09 +/- 0.76 (mean +/- SEM) and propylene glycol: 3.91 +/- 0.71 as compared to dimethyl sulfoxide: 3.49 +/- 0.79 and glycerol: 2.83 +/- 0.40 microm/min per atmosphere. The permeability of chimpanzee sperm in ethylene glycol (2.18 +/- 0.40 x 10(-3) cm/min) and propylene glycol (1.75 +/- 0.17 x 10(-3) cm/min) was higher than in glycerol (1.42 +/- 0.12 x 10(-3) cm/min) and dimethyl sulfoxide (0.82 +/- 0.015 x 10(-3) cm/min). Although chimpanzee sperm tolerated osmotic stress in the range of 169-400 mOsm very well, loss of motility was observed as the solution concentrations diverged from isosmotic condition. Exposure to the four cryoprotectants at 1 M did not cause a significant reduction in sperm motility. This information on membrane permeability characteristics and cryoprotectant tolerance will aid in designing more reliable cryopreservation protocols for chimpanzee sperm.
在22摄氏度下研究了黑猩猩精子的渗透特性。使用电子粒子计数器测定等渗细胞体积,以及暴露于四种常用冷冻保护剂(二甲基亚砜、甘油、丙二醇和乙二醇)后的体积反应。分析数据以确定四种冷冻保护剂的水力传导率和渗透系数。使用暴露于氯化钠溶液的细胞的博伊尔-范特霍夫图来确定非渗透活性体积分数。使用计算机辅助精液分析系统来确定黑猩猩精子的渗透耐受性,以及一步添加和去除1M渗透冷冻保护剂对精子活力的影响。黑猩猩精子的等渗体积为27.7立方微米。非渗透活性细胞分数为69%。与二甲基亚砜(3.49±0.79)和甘油(2.83±0.40)相比,在乙二醇(4.09±0.76,平均值±标准误)和丙二醇(3.91±0.71)存在时水力传导率更高,单位为每分钟每大气压微米。黑猩猩精子在乙二醇(2.18±0.40×10⁻³厘米/分钟)和丙二醇(1.75±0.17×10⁻³厘米/分钟)中的渗透率高于甘油(1.42±0.12×10⁻³厘米/分钟)和二甲基亚砜(0.82±0.015×10⁻³厘米/分钟)。尽管黑猩猩精子在169 - 400毫渗量范围内对渗透应激耐受性良好,但当溶液浓度偏离等渗条件时,观察到活力丧失。在1M浓度下暴露于四种冷冻保护剂不会导致精子活力显著降低。这些关于膜渗透特性和冷冻保护剂耐受性的信息将有助于设计更可靠的黑猩猩精子冷冻保存方案。