Flow cytometric evaluation of ram semen freezability treated with α- tocopherol

Authors : Ongun UYSAL, Muhammed Enes İNANÇ, Ayhan Şükrü GÜNGÖR, Ayhan ATA

Pages : 40-40

View : 19 | Download : 4

Publication Date : 2020-03-31

Article Type : Research Paper

Abstract :The aim of this study was evaluation of ram semen freezability treated with different rate of α-tocopherol in non-breeding season. In this study, ejaculates were collected from four Merino rams insert ignore into journalissuearticles values(2-3 ages were used and belonging to the special sheep farm in Bademli, Burdur/Turkey);. The ejaculates containing spermatozoa with >80% motility and concentrations higher than 1.5x109 spermatozoa/ml insert ignore into journalissuearticles values(normozoospermic); were mixed and used in the study. The mixed ejaculates were divided into five equal aliquots and samples were extended with tris base medium insert ignore into journalissuearticles values(tris, citric acid, glucose); containing 800 μM group, 400 μM group, 200 μM group, 100 μM group or 0 μM group insert ignore into journalissuearticles values(control); α-tocopherol and they were equilibrated to 4°C for 2 hours, frozen in mini straws then stored in liquid nitrogen. Straws were thawed at 37°C for 30s in water bath for plasma membrane acrosome integrity insert ignore into journalissuearticles values(PMAI %); and high mitochondrial membrane potential insert ignore into journalissuearticles values(HMMP %); with flow cytometric analyses. Flow cytometry analyses were performed with a Cytoflex Flow Cytometer insert ignore into journalissuearticles values(Beckman Coulter, Fullerton, CA, USA);. Assessments of semen samples were made with a laser beam at 488 nm insert ignore into journalissuearticles values(50 mW laser output);, with 525 ± 40, and 585 ± 42 and 610 ± 20 nm emission filters. Data were collected from 10.000 events. The highest PMAI insert ignore into journalissuearticles values(24.39±2.11 %); and HMMP insert ignore into journalissuearticles values(39.90±3.13 %); were in 100 μM compared to other groups insert ignore into journalissuearticles values(p<0.05);. Also, the lowest PMAI and HMMP values were obtained in 800 μM and 400 μM groups respectively insert ignore into journalissuearticles values(p<0.05);. In conclusion, 100 μM α-tocopherol should be supplemented to ram semen freezing extender for membrane & acrosome integrity and high mitochondrial membrane potential.
Keywords : α tocopherol, flow cytometry