title: Effects of anti-lipid peroxidases on frozen-thawed boar spermatozoa.
abstract: This study evaluated the effects of anti-lipid peroxidases when supplemented to the thawing and incubation media of frozen-thawed boar spermatozoa. Semen pellets were thawed and incubated in media with 1.0 mM α-tocopherol or diethylenetriamine. After 1 h, the acrosome reaction was induced using calcium ionophore A23187, and acrosomes were evaluated using Wells--Awa staining. The number of spermatozoa with fragmented DNA was evaluated using silver staining after single-cell gel electrophoresis. Membrane lipid peroxidation was measured by the end point generation of malondialdehyde. The diethylenetriamine-supplemented media had a higher (P < 0.05) percentage of acrosome-reacted spermatozoa (84.4 ± 4.1%) compared to the control (78.3 ± 4.2%) and α-tocopherol-supplemented (78.0 ± 3.9%). The control had a higher (P < 0.05) percentage of spermatozoa with fragmented DNA (59.3 ± 4.3%) compared to the DETA (28.7 ± 4.1%) and α-tocopherol supplementation (28.0 ± 3.8%). Spermatozoa supplemented with diethylenetriamine had higher amounts (P < 0.05) of malondialdehyde generated (3.60 ± 0.05 μM/10(7) cells) compared to the α-tocopherol supplementation (0.14 ± 0.05 μM/10(7) cells) and the control (0.12 ± 0.05 μM/10(7) cells). These results indicate that supplementing with either 1.0 mM diethylenetriamine or α-tocopherol during semen thawing and incubation protects against DNA fragmentation, and diethylenetriamine increases the percent of spermatozoa capable of completing the acrosome reaction that could induce membrane lipid peroxidation.
authors.affiliation: Department of Biology, Ferrum College, Ferrum, VA 24088, USA.
authors.forename: Sarah J
authors.initials: SJ
authors.lastname: Casey
authors.forename: Rachel
authors.initials: R
authors.lastname: Taupier
authors.forename: Brian D
authors.initials: BD
authors.lastname: Whitaker
mesh_terms: D020101:Acrosome Reaction / Q000187:drug effects; D000818:Animals; D000975:Antioxidants / Q000494:pharmacology*; D015925:Cryopreservation / Q000379:methods; D053938:DNA Fragmentation; D015227:Lipid Peroxidation / Q000187:drug effects*; D008297:Male; D011073:Polyamines / Q000494:pharmacology*; D012662:Semen Preservation / Q000379:methods*; D013094:Spermatozoa / Q000737:chemistry; D013552:Swine; D024502:alpha-Tocopherol / Q000494:pharmacology*
chemical_list: D000975:Antioxidants; D011073:Polyamines; C005391:diethylenetriamine; D024502:alpha-Tocopherol