Effect of catalase in diluent on survival and acrosome system of boar spermatozoa stored at 4°C

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The effects of catalase with and without reduced glutathione (GSH), ethylenediaminetetraacetate (EDTA) or sodium laurylsulfate (SLS) in diluent on the survival and acrosome system of boar sper-matozoa stored at 4°C were investigated. Semen was collected from 4 Landrace boars by the manual method and only sperm-rich fraction was used. The basic diluent for all experiments contained 20 ml egg yolk, 1.075 g Tris, 0.533 g citric acid, 2.667 g glucose, 100, 000 I.U. potassium penicillin G and 0.1 g dihydrostreptomycin sulfate per 100 ml of diluent. The sperm samples were cooled to room temperature (1823°C) at a constant rate of 0.33°C per min, centrifuged for 1015 min at 600 g and resuspended in the basic diluent to give a concentration of 10 × 108 cells per ml. The sperm suspen-sion was diluted 1:4 (suspension: diluent) with the diluent containing different levels of catalase and/or other agents and gradually cooled to 4°C over 4-h period. After 5, 10 and 15 days of storage at 4°C, sperm motility was assessed microscopically at 15-min intervals during incubation at 37°C for 6090 min. Acrosome morphology of spermatozoa was assessed after staining in a 7.5% (V/V) buffered solution of Giemsa stain. In experiment 1, the effects of 0, 37.5, 75, 150, 300, 600 and 1200 Sigma Units (S.U.) of catalase per ml of diluent on sperm survival were compared. All levels of catalase improved sperm survival during storage for 15 days. The best results were obtained at concen-trations of 751, 200, 1501, 200 and 150600 S.U./ml for spermatozoa stored for 5, 10 and 15 days, respectively. In experiment 2, the effect of different levels of catalase (0600 S.U./ml) on acrosome morphology of spermatozoa were compared. The addition of catalase improved the maintenance of acrosome integrity of spermatozoa and the concentration required for maximum percentage of sper-matozoa with normal acrosome was 150600 S.U./ml. Unusual stainability of the anterior boundary of acrosome was the major morphological change observed following storage, and the percentage of spermatozoa with such damage was decreased with catalase levels higher than 75 S.U./ml. The results of these experiments suggest that a catalase level of 150300 S.U./ml is optimal for preserv-ing boar spermatozoa under conditions employed. In three other experiments, the effects of catalase (300 S.U./ml) in diluent containing different levels of GSH (0, 5, 10 and 20 mM), EDTA (0, 2.5, 5 and 10 mM) or SLS (0, 0.02, 0.04, 0.08 and 0.16% W/V) on sperm survival were examined. Again, the addition of catalase improved sperm survival. GSH and SLS also prolonged sperm life when they were used singly, but the beneficial effect of these agents was reduced more rapidly than that of catalase alone. Furthermore, both GSH and SLS had no beneficial effect when they were combined with catalase. EDTA was not beneficial to sperm survival regardless of the presence or absence of catalase.
日本繁殖生物学会の論文

Effect of catalase in diluent on survival and acrosome system of boar spermatozoa stored at 4°C

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日本繁殖生物学会 | 論文

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