Utilisation of sperm-binding assay combined with computer-assisted sperm analysis to evaluate frozen-thawed bull semen.
Due to homologies between the chicken egg perivitelline membrane with mammalian zona pellucida proteins, spermatozoa of several species are able to bind to this membrane. However, adequate standardisation is required to attest possible applications of this technique for semen evaluation of a given species. Therefore, we thawed and divided cryopreserved semen samples into two aliquotes, one kept in water bath at 37 °C (thawed) and the other submitted to snap-freezing to damage sperm cells (dead spermatozoa). Aliquotes were mixed into different ratios of thawed:dead cells and analysed for motility, membrane and acrosomal integrity, and mitochondrial activity. In parallel, chicken egg perivitelline membranes were inseminated with these ratios, and the number of spermatozoa bound per mm(2) of membrane was assessed by conventional microscopy (CM) and computer-assisted sperm analysis (CASA). Linear regression showed high correlation between thawed:dead sperm ratio and number of spermatozoa bound to the membrane (CM: r(2) = 0.91 and CASA: r(2) = 0.92 respectively). Additionally, positive correlations were found between the number of spermatozoa bound to the membrane and acrosomal integrity, membrane integrity, mitochondrial activity and motility. These findings indicate that sperm-egg-binding assay associated with CASA is a reliable, practical and inexpensive method for examining the fertilising capacity of cryopreserved bull semen.
['Losano|J D A|JD|', 'Angrimani|D S R|DS|', 'Pereira|R J G|RJ|', 'Rocha|A M|AM|', 'Criscuolo|T S|TS|', 'Barnabe|V H|VH|', 'Barnabe|R C|RC|', 'Mendes|C M|CM|', 'Assumpção|M E O A|ME|', 'Nichi|M|M|']
D000818:Animals D002417:Cattle D002645:Chickens D015925:Cryopreservation D003936:Diagnosis, Computer-Assisted D004531:Eggs D008297:Male D055101:Semen Analysis D012662:Semen Preservation D013084:Sperm-Ovum Interactions D015044:Zona Pellucida
sperm-binding assay Bull perivitelline membrane semen analysis