The spermatozoa of the dasyurid marsupial, Sminthopsis crassicaudata, are highly susceptible to cold shock

Abstract

Since the late 1970s research has suggested that marsupial spermatozoa did not suffer cold shock. We have re-examined cold shock to investigate problems with freezing of spermatozoa from a dasyurid marsupial, the fat-tailed dunnart (Sminthopsis crassicaudata). Epididymal spermatozoa were rapidly cooled to 0.5°C in a pre-cooled tube held in an iced-water slurry. Upon re-warming all spermatozoa were immotile and the addition of 10% or 20% egg yolk to the sperm medium had no beneficial effect. Spermatozoa that were rapidly cooled to 4°C maintained only 2% motility when re-warmed but the addition of at least 10% egg yolk was beneficial and upon re-warming greater than 65% of the initial motility was maintained. In order to achieve motile spermatozoa at 0°C, controlled-rate cooling at 0.5°C min–1 was examined. In the absence of egg yolk there was a significant decline in the percentage of motile spermatozoa below 4°C. However, the inclusion of at least 10% egg yolk resulted in no loss of motility in spermatozoa cooled to 0°C. This is the first experimental study indicating that spermatozoa from a marsupial are highly susceptible to cold shock and that the impact of rapid chilling can be mitigated by the addition of 10% egg yolk. The ability to successfully cool the spermatozoa of S. crassicaudata to 0°C may have an important role in future studies examining dasyurid sperm cryopreservation.