Hypoosmotic swelling of boar spermatozoa compared to other methods for analysing the sperm membrane

The aims of this work were to adapt the hypoosmotic swelling test (HOST) to boar spermatozoa and to compare this method with other tests which evaluate the integrity of the sperm membrane. The spermatozoa were incubated in 50, 100, 150 or 200 mOsm/L solutions for 5, 30, 60 or 120 min. An easily identifiable swelling and coiling of the tails occurred when the boar spermatozoa were incubated at 37 degrees C for 30 to 120 min in a mixture of fructose and Na-citrate (100-150 mOsm/L). Transmission electron microscopy showed that the hypoosmotic swelling reaction of the spermatozoa was caused by coiling of the flagellum inside the plasma membrane. When used as described, HOST was found to be highly reliable when known populations of live spermatozoa were tested. We also compared the results obtained with HOST with those obtained using eosin Y and carboxyfluorescein diacetate. The percentage of spermatozoa unstained with eosin Y and the percentage of spermatozoa which fluoresced with carboxyfluorescein diacetate were similar. However, the hypoosmotic swelling values were significantly below those of the other tests. This may be because either HOST evaluates different aspects of sperm membrane than other sperm membrane tests or the membranes of some spermatozoa are inactivated by contact with the hypoosmotic solution. In short, our findings suggest that HOST is a sensitive and reproducible test to assess the functional integrity of boar sperm membranes after incubation under hypoosmotic stress conditions and may be a useful tool for detecting subpopulations of subviable spermatozoa when used in conjunction with another type of membrane integrity test.