Determination of Plasma Membrane Characteristics of Boar Spermatozoa and their Relevance to Cryopreservation1

Abstract

The osmotic tolerance limits for boar spermatozoa were determined at 22 degrees C. These cells can swell to within 1.02 times and shrink to within 0.97 times their isosmotic volume and maintain > 70% motility. In the presence of an extender, cells can swell to within 1.1 times and shrink to within 0.97 times their isosmotic volume and maintain > 70% motility. Plasma membrane permeability coefficients were determined in the presence of 1 M dimethyl sulfoxide (DMSO), 1 M glycerol, and 2 M ethylene glycol (EG) at 22 degrees C. Hydraulic conductivity (Lp) was estimated to be 0.120+/-0.016 (mean+/-SEM), 0.138+/-0.006, and 0.204 +/-0.021 microm/min/atm in the presence of DMSO, glycerol, and EG, respectively, at 22 degrees C. Solute permeability (P[CPA]) was determined to be 0.930+/-0.118, 0.481+/-0.045, and 1.98+/-0.106 x 10(-3) cm/min, for DMSO, glycerol, and EG, respectively. Subsequent experiments were performed at 8 degrees C and 0 degrees C. Activation energies were calculated for Lp in the presence of glycerol and EG to be 7.20 and 11.51 Kcal/mol, respectively. The activation energies for P(CPA) were 4.06 and 7.48 Kcal/mol for glycerol and EG permeability, respectively. These membrane characteristics were used to calculate volume flux during addition and removal of cryoprotectant agents as well as during cooling and warming. In addition, the potential for intracellular ice formation during cooling and warming was calculated.