Cryoprotectant and freezing-process alter the ability of chicken sperm to acrosome react

Sperm surviving after freezing-thawing is usually 40-50% of the initial population. Damage during this process affects both fertilizing ability and its duration in avian species. However, the effect of cryopreservation on the sperm ability to undergo the acrosome reaction, the initial event of fertilization, is still in question in birds. In this paper, the influence of cryoprotectant (glycerol and dimethylacetamide-DMA) and of two different cryopreservation processes (pellets or straws) on the ability of rooster sperm to acrosome react (AR measured with PNA-FITC) was studied. Motility parameters (CASA) and plasma membrane integrity (propidium iodide exclusion) were also measured. The addition of cryoprotectants (CPA) immediately provoked a dramatic decrease in the ability of sperm to undergo the acrosome reaction, glycerol being more harmful than DMA. The cryoprotectant removal was also harmful. The other parts of the freezing process further decreased the ability to AR. Motility was affected to a lesser extent by CPA presence although plasma membrane integrity was not altered. The DMA/rapid freezing procedure was the most harmful for plasma membrane integrity. Taken together, these results show that AR is more dramatically altered by CPA presence than motility and membrane integrity and CPA provokes a more pronounced effect than the freezing-thawing process especially in the case of using glycerol/slow freezing process.