Significant variability among bulls in the sperm membrane permeability for water and glycerol: possible implications for semen freezing protocols for individual males.

The aim of this study was to test the hypothesis that bulls have significant intra-individual differences in the hydraulic conductivity (L(p)) and permeability coefficient for glycerol (P(s)) of the sperm cell membrane. The permeability parameters were determined at 22, 10, and 0 degrees C of sperm from 7 Holstein Frisian artificial insemination (AI) bulls, using four ejaculates per bull. A stopped-flow approach was applied to provide temporal resolution sufficient to measure rapid cell volume changes under anisosmotic conditions in the absence or presence of glycerol. This technique utilizes a concentration-dependent self-quenching entrapped fluorophore. The resulting cell volume changes were used in three-parameter fitting calculations to compute L(p) in the absence glycerol, and L(p) in the presence of glycerol (L(p)(gly)) and P(s). Averaged over all bulls, L(p) in the absence of glycerol was 0.28+/-0.01, 0.15+/-0.01 and 0.10+/-0.01 microm min(-1)atm(-1) (mean+/-SD) at 22, 10 and 0 degrees C, respectively, yielding an Arrhenius activation energy (E(a)) of 7.39 kcal/mol. The average L(p)(gly) value at 22 degrees C, was 3.8 times lower than L(p) in the absence of glycerol (P<0.05). L(p)(gly), P(s), and the reflection coefficient (sigma) at 22 degrees C were 0.073+/-0.015 microm min(-1)atm(-1), 0.80+/-0.33 x 10(-3)cm min(-1), and 0.92+/-0.10 (mean+/-SD), respectively. Subsequent experiments were performed at 10 and 0 degrees C. Activation energies for L(p)(gly) and P(s) were 10.08 and 8.77 kcal/mol, respectively. The significant differences between individual bulls in L(p) and P(s) indicate that individual males may require individual adjustments of the cooling protocol. Application of these data in a theoretical model to simulate the osmotic events during freezing resulted in predicted optimal cooling rates in the range of published empirical values.