The ability to reverse swelling caused by hypo-osmotic stress is an important cell function; in spermatozoa, it is likely to be of consequence during ejaculation and also during the thawing process that terminates cryopreservation. In this study, the time course of boar and bull sperm volume changes after exposure to hypo-osmotic conditions at 39 degrees C was recorded. Cell volume measurements of washed sperm suspensions were performed electronically in Hepes-buffered saline solutions of 300 and 180 mosmol kg(-1) containing 2.5 mmol K(+) l(-1). Treatment with quinine in the presence or absence of the potassium ionophore valinomycin was used to determine whether potassium channels were involved in the reversal of swelling. After exposure to hypo-osmotic conditions, both bull and boar spermatozoa showed initial swelling (up to 200% and 140% of initial volume, respectively, within 5 min), which was subsequently partially reversed (to about 150% and 120%, respectively, after 20 min). Incubation with quinine led to an increase in swelling in both species. However, bull sperm volume was already maximal (up to 294%) after 30 s and declined thereafter, whereas boar sperm volume increased slowly to a maximum of about 220% after 20 min. Valinomycin treatment caused quinine-induced swelling in bull spermatozoa to decrease rapidly to control (no quinine, no valinomycin) values, whereas in quinine-treated boar spermatozoa it had an opposite, enhancing effect. Interpreting these results in the light of data from studies by others on a variety of cell types, it is proposed that swelling-activated potassium channels are involved in regulatory volume decrease in both species of spermatozoa, but that boar spermatozoa may contain fewer swelling-activated chloride channels than do bull spermatozoa.
AM Petrunkina, RA Harrison, M Hebel, KF Weitze and E Topfer-Petersen
AM Petrounkina, RA Harrison, R Petzoldt, KF Weitze and E Topfer-Petersen
The osmotic reactivity of boar spermatozoa during incubation in vitro was studied using a hypo-osmotic swelling test in conjunction with electronic measurement of cell volume. Sperm populations showed fluctuations in both iso-osmotic cell volume and hypo-osmotic volume response that fitted mathematical models for periodicity. Significant differences of frequency and amplitude were observed during sperm incubation under capacitating conditions as compared with those under non-capacitating conditions. In addition, different boars showed specific differences in their fluctuation characteristics under capacitating conditions. During incubation under capacitating conditions, a decrease in osmotic reactivity was observed that correlated with a decrease in motility, while the absolute value of the earliest maximum of the osmotic-induced response correlated with an increase in the proportion of discharged acrosomes. The time course of the cyclical behaviour of osmotic reactivity may be a useful parameter for assessing boar sperm response to capacitating conditions.