Escherichia coli membrane-derived oxygen-reducing enzyme system (Oxyrase) protects bubaline spermatozoa during cryopreservation
© 2020 Wiley Periodicals LLC..
The objective of this study was to determine the effectiveness of deoxygenation of semen extender using Escherichia coli membrane-derived oxygen scavenger (Oxyrase) on post-thaw quality of buffalo (Bubalus bubalis) spermatozoa. Sixteen semen ejaculates, four each from four bulls, were each divided into five equal fractions, diluted using Tris-egg yolk extender supplemented with different concentrations of Oxyrase (0, 0.3, 0.6, 0.9, and 1.2 U/ml), designated as treatments T1, T2, T3, T4, and T5, respectively, and cryopreserved. Immediately after thawing, Oxyrase did not improve sperm kinetics and motility; however, it improved the keeping quality (significantly lower deterioration of post-thaw sperm motility after incubation for 120 min) in T3. Further, T3 reduced (p < .05) cholesterol efflux and protected the intactness of the sperm plasma membrane. Flow cytometry with Fluo-3 AM/propidium iodide (PI) dual staining revealed the highest (p < .05) proportion of live spermatozoa with low intracellular calcium in T3. Oxyrase supplementation protected spermatozoa from premature capacitation which was confirmed by low expression of tyrosine-phosphorylated proteins (32, 75, and 80 kDa) and a relatively lower percentage of F-pattern (uncapacitated spermatozoa) in chlortetracycline assay. Importantly, the Oxyrase fortification decreased superoxide anion in a dose-dependent manner indicating reduced availability of oxygen at sperm mitochondrial level. Similarly, in Oxyrase-fortified sperm, malondialdehyde concentration, an index of lipid peroxidation, is also reduced in a dose-dependent manner. In conclusion, we demonstrate that deoxygenation of buffalo semen by Oxyrase has the potential of improving post-thaw sperm quality by overcoming the problem of cryocapacitation and oxidative damage during cryopreservation process.
| Medienart: |
E-Artikel |
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| Erscheinungsjahr: |
2020 |
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| Erschienen: |
2020 |
| Enthalten in: |
Zur Gesamtaufnahme - volume:87 |
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| Enthalten in: |
Molecular reproduction and development - 87(2020), 10 vom: 07. Okt., Seite 1048-1058 |
| Sprache: |
Englisch |
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| Beteiligte Personen: |
Dalal, Jasmer [VerfasserIn] |
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| Links: |
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| Themen: |
Buffalo bull |
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| Anmerkungen: |
Date Completed 26.08.2021 Date Revised 26.08.2021 published: Print-Electronic Citation Status MEDLINE |
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| doi: |
10.1002/mrd.23411 |
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| funding: |
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| Förderinstitution / Projekttitel: |
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NLM313553297 |
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| 245 | 1 | 0 | |a Escherichia coli membrane-derived oxygen-reducing enzyme system (Oxyrase) protects bubaline spermatozoa during cryopreservation |
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| 520 | |a The objective of this study was to determine the effectiveness of deoxygenation of semen extender using Escherichia coli membrane-derived oxygen scavenger (Oxyrase) on post-thaw quality of buffalo (Bubalus bubalis) spermatozoa. Sixteen semen ejaculates, four each from four bulls, were each divided into five equal fractions, diluted using Tris-egg yolk extender supplemented with different concentrations of Oxyrase (0, 0.3, 0.6, 0.9, and 1.2 U/ml), designated as treatments T1, T2, T3, T4, and T5, respectively, and cryopreserved. Immediately after thawing, Oxyrase did not improve sperm kinetics and motility; however, it improved the keeping quality (significantly lower deterioration of post-thaw sperm motility after incubation for 120 min) in T3. Further, T3 reduced (p < .05) cholesterol efflux and protected the intactness of the sperm plasma membrane. Flow cytometry with Fluo-3 AM/propidium iodide (PI) dual staining revealed the highest (p < .05) proportion of live spermatozoa with low intracellular calcium in T3. Oxyrase supplementation protected spermatozoa from premature capacitation which was confirmed by low expression of tyrosine-phosphorylated proteins (32, 75, and 80 kDa) and a relatively lower percentage of F-pattern (uncapacitated spermatozoa) in chlortetracycline assay. Importantly, the Oxyrase fortification decreased superoxide anion in a dose-dependent manner indicating reduced availability of oxygen at sperm mitochondrial level. Similarly, in Oxyrase-fortified sperm, malondialdehyde concentration, an index of lipid peroxidation, is also reduced in a dose-dependent manner. In conclusion, we demonstrate that deoxygenation of buffalo semen by Oxyrase has the potential of improving post-thaw sperm quality by overcoming the problem of cryocapacitation and oxidative damage during cryopreservation process | ||
| 650 | 4 | |a Journal Article | |
| 650 | 4 | |a Research Support, Non-U.S. Gov't | |
| 650 | 4 | |a Oxyrase | |
| 650 | 4 | |a buffalo bull | |
| 650 | 4 | |a cryopreservation | |
| 650 | 4 | |a oxidative stress | |
| 650 | 4 | |a spermatozoa | |
| 650 | 7 | |a Cryoprotective Agents |2 NLM | |
| 650 | 7 | |a Oxygenases |2 NLM | |
| 650 | 7 | |a EC 1.13.- |2 NLM | |
| 650 | 7 | |a Oxyrase |2 NLM | |
| 650 | 7 | |a EC 1.14.- |2 NLM | |
| 700 | 1 | |a Chandolia, Ramesh Kumar |e verfasserin |4 aut | |
| 700 | 1 | |a Jan, Mustafa Hassan |e verfasserin |4 aut | |
| 700 | 1 | |a Pawaria, Shikha |e verfasserin |4 aut | |
| 700 | 1 | |a Verma, Nisha |e verfasserin |4 aut | |
| 700 | 1 | |a Jerome, Andonissamy |e verfasserin |4 aut | |
| 700 | 1 | |a Kumar, Dharmendra |e verfasserin |4 aut | |
| 700 | 1 | |a Kumar, Pradeep |e verfasserin |4 aut | |
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| 773 | 1 | 8 | |g volume:87 |g year:2020 |g number:10 |g day:07 |g month:10 |g pages:1048-1058 |
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