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標題: Hydrostatic pressure pre-treatment affects the protein profile of boar sperm before and after freezing-thawing
作者: Huang, S.Y.
Pribenszky, C.
Kuo, Y.H.
Teng, S.H.
Chen, Y.H.
Chung, M.T.
Chiu, Y.F.
關鍵字: Boar;Frozen semen;Hydrostatic pressure;Protein profile;spermatogenic cells;polyacrylamide-gels;escherichia-coli;stress-response;shock proteins;mouse;identification;dehydrogenase;resistance;expression
Project: Animal Reproduction Science
期刊/報告no:: Animal Reproduction Science, Volume 112, Issue 1-2, Page(s) 136-149.
A sublethal environmental stress, high-hydrostatic pressure (HHP) was reported to significantly improve the motility, viability and fertility parameters of frozen bull and boar semen. However, the mechanism of how HHP treatment improves survival rates at sperm cryopreservation remains unclear. The purpose of this study was to evaluate the effect of HHP treatment of fresh boar semen on the protein profile of boar sperm before and after freezing. Fresh, extended semen of eight boars was split, one part was treated with 200, 300 or 400 bar for 90 min using a custom made pressuring device before the start of the semen freezing procedure, and the other part was prepared without HHP treatment. After thawing, samples were checked for motility. The effect of HHP treatment on the post-thaw motility of frozen semen was significant (P = 0.02). Post-thaw motility of each treatment groups increased compared to control (46% vs. 52%, 56% and 56%; control vs. 200 bar, 300 bar and 400 bar treatments). Samples for protein analysis were collected from the 300 bar treatment group before HHP treatment at room temperature (25 +/- 3 degrees C, at 5 degrees C of the cooling process and after thawing with or without HHP treatment. The sperm were lysed using a urea-pyranoside-dithiothreitol buffer to extract their proteins for protein analysis. Approximately 800 mu g total proteins were assayed by two-dimensional gel electrophoresis and stained with colloidal Coomassie blue. The levels of 125 protein spots were quantified. The results revealed that the levels of 7 protein spots differed significantly among treatments. The identities of various protein constituents were identified by mass spectrometry and database searching. Ubiquinol-cytochrome c reductase complex core protein 1, perilipin, and carbohydrate-binding protein AWN precursor were identified as HHP response proteins being significantly higher in HHP-treated samples. Testis-specific glyceraldehyde 3-phosphate dehydrogenase, outer dense fiber of sperm tails 2 isoform 10, cytosolic 5'-nucleotidase 1B, and quinone oxidoreductase represented the cooling and freezing related proteins. The differing levels of these identified proteins could be valuable for further exploring the protective mechanism of the HHP treatment in frozen-thawed porcine sperm. (C) 2008 Elsevier B.V. All rights reserved.
ISSN: 0378-4320
DOI: 10.1016/j.anireprosci.2008.04.016
Appears in Collections:動物科學系

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