Studies were designed to evaluate bull sperm motility and membrane int
egrity following exposure to Tyrode's solution varying in osmolality f
rom 100 to 1537 mOsm. Congo red and bisbenzimide (HOECHST 33258) stain
s were used to distinguish between sperm with intact versus disrupted
plasma membranes. Sperm motility was subjectively evaluated. No signif
icant differences mere found between motile and unstained sperm (sperm
with intact membranes) in solutions with nearly physiologic osmolalit
ies between 200 and 300 mOsm. However, in 100- and 150-mOsm solutions,
the percentage of motile sperm (5 and 19%, respectively) was lower th
an the percentage of sperm unstained with Congo red (18 and 35%). With
HOECHST 33258 stain, the corresponding values were 7 and 14% versus 2
6 and 31%. The percentage of motile sperm declined greatly in the 500-
mOsm medium, but the proportion of unstained sperm was affected Little
until the osmotic pressure exceeded 732 mOsm. Partial recovery of mot
ility occurred when sperm were returned to the isotonic medium. This s
tudy indicated that the sperm plasma membrane was more resistant to os
motic damage than were the mechanisms responsible for sperm motility.
Lack of motility in hypertonic media was not an absolute indicator of
cell death, and unstained sperm overestimated sperm viability, which a
re factors to consider when sperm are being evaluated, especially rela
tive to cryopreservation.