Experiment lc exposed cells to a narrower osmotic range of anisosmotic conditions. Exposure to anisosmotic conditions was performed as described above using the following osmolalities of PBS: 140, 220, 260, 280, 290, 300, 320, 360, 440, and 600 mOsmol/kg. After mixing, the suspension was allowed to equilibrate for 10 min before motility was assessed. A total of three boars (n = 3) were used and all data were collected at 22°C.
Experiment Id was designed for return to isosmolality after exposure to anisosmotic conditions. After exposure, cells were returned to near isosmolality by adding 1500 |xl of isosmotic PBS. Motility was assessed after 1 min and after 5 min. buy asthma inhalers
Experiment 2. Activation energy for boar spermatozoa solute and water permeability. One hundred microliters of 2 M glycerol, 2 M DMSO, or 4 M EG was added drop by drop over 60 sec to 100 (xl of cell suspension, yielding final CPA concentrations of 1 M and 2 M, respectively. Cells were allowed to equilibrate for approximately 3 min before all 200 |xl were returned to isosmotic media. Changes in cell volume were measured over time. A total of 3 donors (n = 3) were used, and each experimental run was replicated three times for each donor. The experiments were performed at 22°C, 8°C, and 0°C in the presence of glycerol and EG, and at 22°C for DMSO.
Experiment 3. Osmotic tolerance limits of boar spermatozoa in the presence of extender. The same procedures were performed as described in experiment lc; however, the spermatozoa were suspended approximately 1:3 in Modena extender.