Osmotic tolerance and membrane permeability characteristics of rhesus monkey (Macaca mulatta) spermatozoa.

Biophysical characteristics of the plasma membrane, such as osmotic sensitivity and water and cryoprotectant permeability are important determinants of the function of spermatozoa after cryopreservation. A series of experiments was conducted with rhesus macaque spermatozoa at 23 degrees C to determine their: (1) cell volume and osmotically inactive fraction of the cell volume; (2) permeability coefficients for water and the cryoprotectants dimethyl sulfoxide, glycerol, propylene glycol, and ethylene glycol; (3) tolerance to anisosmotic conditions; and (4) motility after a one step addition and removal of the four cryoprotectants. An electronic particle counter and computer aided semen analysis were used to determine the cell volume and permeability coefficients, and motility, respectively. Rhesus spermatozoa isosmotic cell volume was 27.7+/-3.0 microm3 (mean+/-SEM) with an osmotically inactive cell fraction of 51%. Hydraulic conductivity in the presence of dimethyl sulfoxide, glycerol, propylene glycol, and ethylene glycol was 1.09+/-0.30, 0.912+/-0.27, 1.53+/-0.53, and 1.94+/-0.47 microm/min/atm, respectively. Cryoprotectant permeability was 1.39+/-0.31, 2.21+/-0.32, 3.38+/-0.63, and 6.07+/-1.1 (x10(-3)cm/min), respectively. Rhesus sperm tolerated all hyposmotic exposures. However, greater than 70% motility loss was observed after exposure to solutions of 600 mOsm and higher. A one step addition and removal of all four cryoprotectants did not cause significant motility loss. These data suggest that rhesus sperm are tolerant to hyposmotic conditions, and ethylene glycol may be the most appropriate cryoprotectant for rhesus sperm cryopreservation, as it has the highest permeability coefficient of the tested cryoprotectants.

Chimpanzee (Pan troglodytes) spermatozoa osmotic tolerance and cryoprotectant permeability characteristics.

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.