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KMID : 0900920030270020153
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Korean journal of Animal Reproduction
2003 Volume.27 No. 2 p.153 ~ p.161
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Changes of Glycosidase Activity and Fertilizing Ability in Vitro by Incubation of Frozen-Thawed Spermatozoa in the Pig
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Hwang In-Sun
Cheong Hee-Tae
Yang Boo-Keun
Kim Choung-Ik
Park Choon-Keun
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Abstract
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This study has evaluated effect of the spermatozoa incubation on the glycosidase activity and fertilizing ability in vitro in the pig. To identify sperm glycosidases specific for sugar residues found in the zona pellucida of pig oocytes, the spermatozoa were treated experimentally and assayed for activities of alpha-L-fucosidase, alpha-D-mannosidase, beta-D-galactosidase and N-acetyl-beta-D-glucosaminidase (beta-GlcNAc’ase). The glycosidases activity were higher in spermatozoa incubated for 2h than without incubation. The beta-GlcNAc’ase activity was at least two-fold higher than other glycosidase regardless of spermatozoa incubation. In the same glycosidases, the activity had a tendency to increase as time of spermatozoa incubation was prolonged, but there were no differences in spermatozoa incubated during the various periods (4~24h). The percentages of spermatozoa that reached acrosome reaction were affected by glycosidases in the medium (P<0.05, for mannosidase), and were higher in spermatozoa with that than without incubation. On the other hand, the spermatozoa motility were decreased with incubation periods, but no effects by different glycosidases on the change of sperm motility during the various periods of incubation. In other experiment, the binding and penetration of pig spermatozoa were tested with oocytes matured in vitro in the presence of various glycosidase. The penetration rates were decreased with incubation of spermatozoa when oocytes were inseminated in medium with different glycosidases. These rates were higher in spermatozoa non-incubated than with incubation for 2h (P<0.05 for GlcNAc’ase; P<0.01 for control group). The sperm-zona binding rate in control group were higherthan in medium with glycosidases. In addition, the highest binding rate were obtained in medium with GlcNAc’ase. In all glycosidases, the sperm-zona binding rate in spermatozoa without incubation were higher than incubation for 2h. The significant differences were obtained in spermatozoa treated with alpha-D-mannosidase (P<0.05). These results suggest that beta-GlcNAc’ase is present mainly in the plasma membrane of pig spermatozoa. It was also shown that the glycosidase activity were increased in all glycosidases in spite of low sperm-zona binding rate and penetration rates by spermatozoa incubation.
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KEYWORD
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Fertilization ability, Glycosidase activity, In-vitro, Pig, Spermatozoa incubation
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