Preliminary Study of Effects of Centrifugation of Stallion Semen on Motility and Pro-AKAP4 Concentration over 72 Hours Post-collection

摘要

The study aimed to investigate the effects of centrifugation of stallion semen on progressive sperm motility over a 72-hour period;and determine if the protein marker,precursor of A-kinase anchor protein 4(pro-AKAP4),may be maintained at a higher level over time when compared to non-centrifuged samples.Pro-AKAP4 has been studied for its function in stabilizing the structure of spermatozoa and its use as a marker for fertilizing capacity of spermatozoa.Semen collections were obtained from four fertile,commercially used Standardbred stallions collected three times each.Semen samples were diluted to a final concentration of 50 million total cells per milliliter.One extended aliquot of semen from each collection was used for the non-centrifuged treatment,and one extended aliquot of semen was centrifuged at 750×g for 10 min.The supernatant from the centrifuged sample was then removed with a 10 mL plastic transfer pipette,and the spermatozoa cell pellet was suspended in fresh extender to a final concentration of 50 million total cells per milliliter.Both centrifuged and non-centrifuged samples from each collection were stored in Equine Express II stallion semen shipping containers for 72 h and fresh ice packs were added to each container every 24 h.Both progressive sperm motility and levels of pro-AKAP4 were determined at times 0,12,24,48 and 72 h after collection.Differences were found between stallions,times and treatments with the main finding being that pro-AKAP4 concentration in centrifuged samples was maintained at a much higher level over the 72-hour period studied.