Caspase Activation, Hydrogen Peroxide Production and Akt Dephosphorylation Occur During Stallion Sperm Senescence
To investigate the mechanisms inducing sperm death after ejaculation, stallion ejaculates were incubated in BWW media during 6 h at 37°C. At the beginning of the incubation period and after 1, 2, 4 and 6 h sperm motility and kinematics (CASA), mitochondrial membrane potential and membrane permeabili...
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| Veröffentlicht in: | Reproduction in domestic animals 2014-08, Vol.49 (4), p.657-664 |
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| container_title | Reproduction in domestic animals |
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| creator | Gallardo Bolaños, JM Balao da Silva, C Martín Muñoz, P Plaza Dávila, M Ezquerra, J Aparicio, IM Tapia, JA Ortega Ferrusola, C Peña, FJ |
| description | To investigate the mechanisms inducing sperm death after ejaculation, stallion ejaculates were incubated in BWW media during 6 h at 37°C. At the beginning of the incubation period and after 1, 2, 4 and 6 h sperm motility and kinematics (CASA), mitochondrial membrane potential and membrane permeability and integrity were evaluated (flow cytometry). Also, at the same time intervals, active caspase 3, hydrogen peroxide, superoxide anion (flow cytometry) and Akt phosphorylation (flow cytometry) were evaluated. Major decreases in sperm function occurred after 6 h of incubation, although after 1 h decrease in the percentages of motile and progressive motile sperm occurred. The decrease observed in sperm functionality after 6 h of incubation was accompanied by a significant increase in the production of hydrogen peroxide and the greatest increase in caspase 3 activity. Additionally, the percentage of phosphorylated Akt reached a minimum after 6 h of incubation. These results provide evidences that sperm death during in vitro incubation is largely an apoptotic phenomena, probably stimulated by endogenous production of hydrogen peroxide and the lack of prosurvival factors maintaining Akt in a phosphorylated status. Disclosing molecular mechanisms leading to sperm death may help to develop new strategies for stallion sperm conservation. |
| doi_str_mv | 10.1111/rda.12343 |
| format | Article |
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At the beginning of the incubation period and after 1, 2, 4 and 6 h sperm motility and kinematics (CASA), mitochondrial membrane potential and membrane permeability and integrity were evaluated (flow cytometry). Also, at the same time intervals, active caspase 3, hydrogen peroxide, superoxide anion (flow cytometry) and Akt phosphorylation (flow cytometry) were evaluated. Major decreases in sperm function occurred after 6 h of incubation, although after 1 h decrease in the percentages of motile and progressive motile sperm occurred. The decrease observed in sperm functionality after 6 h of incubation was accompanied by a significant increase in the production of hydrogen peroxide and the greatest increase in caspase 3 activity. Additionally, the percentage of phosphorylated Akt reached a minimum after 6 h of incubation. These results provide evidences that sperm death during in vitro incubation is largely an apoptotic phenomena, probably stimulated by endogenous production of hydrogen peroxide and the lack of prosurvival factors maintaining Akt in a phosphorylated status. Disclosing molecular mechanisms leading to sperm death may help to develop new strategies for stallion sperm conservation.</description><identifier>ISSN: 0936-6768</identifier><identifier>EISSN: 1439-0531</identifier><identifier>DOI: 10.1111/rda.12343</identifier><identifier>PMID: 24924976</identifier><language>eng</language><publisher>Germany: Blackwell Science</publisher><subject>Animal reproduction ; Animals ; Apoptosis ; Caspase 3 - metabolism ; Caspase 7 - metabolism ; caspases ; Caspases - metabolism ; Cell Membrane Permeability - physiology ; Cellular Senescence - physiology ; death ; dephosphorylation ; ejaculation ; Enzyme Activation ; Enzymes ; flow cytometry ; Flow Cytometry - veterinary ; Horses ; Horses - physiology ; Hydrogen peroxide ; Hydrogen Peroxide - metabolism ; hydrogen production ; incubation period ; kinematics ; Male ; membrane permeability ; membrane potential ; Membrane Potential, Mitochondrial - physiology ; mitochondrial membrane ; Phosphorylation ; Proto-Oncogene Proteins c-akt - metabolism ; Semen Preservation - veterinary ; Senescence ; Sperm ; sperm motility ; Sperm Motility - physiology ; spermatozoa ; Spermatozoa - physiology ; Spermatozoa - ultrastructure ; stallions ; superoxide anion ; Time Factors</subject><ispartof>Reproduction in domestic animals, 2014-08, Vol.49 (4), p.657-664</ispartof><rights>2014 Blackwell Verlag GmbH</rights><rights>2014 Blackwell Verlag GmbH.</rights><rights>Copyright © 2014 Wiley-VCH Verlag GmbH & Co. KGaA Weinheim</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c4483-78cb3315445390d50e11850a8c46a7440ef243939c7b8e3128fe6db386b5fcb93</citedby><cites>FETCH-LOGICAL-c4483-78cb3315445390d50e11850a8c46a7440ef243939c7b8e3128fe6db386b5fcb93</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://onlinelibrary.wiley.com/doi/pdf/10.1111%2Frda.12343$$EPDF$$P50$$Gwiley$$H</linktopdf><linktohtml>$$Uhttps://onlinelibrary.wiley.com/doi/full/10.1111%2Frda.12343$$EHTML$$P50$$Gwiley$$H</linktohtml><link.rule.ids>314,780,784,1417,27924,27925,45574,45575</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/24924976$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Gallardo Bolaños, JM</creatorcontrib><creatorcontrib>Balao da Silva, C</creatorcontrib><creatorcontrib>Martín Muñoz, P</creatorcontrib><creatorcontrib>Plaza Dávila, M</creatorcontrib><creatorcontrib>Ezquerra, J</creatorcontrib><creatorcontrib>Aparicio, IM</creatorcontrib><creatorcontrib>Tapia, JA</creatorcontrib><creatorcontrib>Ortega Ferrusola, C</creatorcontrib><creatorcontrib>Peña, FJ</creatorcontrib><title>Caspase Activation, Hydrogen Peroxide Production and Akt Dephosphorylation Occur During Stallion Sperm Senescence</title><title>Reproduction in domestic animals</title><addtitle>Reprod Dom Anim</addtitle><description>To investigate the mechanisms inducing sperm death after ejaculation, stallion ejaculates were incubated in BWW media during 6 h at 37°C. At the beginning of the incubation period and after 1, 2, 4 and 6 h sperm motility and kinematics (CASA), mitochondrial membrane potential and membrane permeability and integrity were evaluated (flow cytometry). Also, at the same time intervals, active caspase 3, hydrogen peroxide, superoxide anion (flow cytometry) and Akt phosphorylation (flow cytometry) were evaluated. Major decreases in sperm function occurred after 6 h of incubation, although after 1 h decrease in the percentages of motile and progressive motile sperm occurred. The decrease observed in sperm functionality after 6 h of incubation was accompanied by a significant increase in the production of hydrogen peroxide and the greatest increase in caspase 3 activity. Additionally, the percentage of phosphorylated Akt reached a minimum after 6 h of incubation. These results provide evidences that sperm death during in vitro incubation is largely an apoptotic phenomena, probably stimulated by endogenous production of hydrogen peroxide and the lack of prosurvival factors maintaining Akt in a phosphorylated status. Disclosing molecular mechanisms leading to sperm death may help to develop new strategies for stallion sperm conservation.</description><subject>Animal reproduction</subject><subject>Animals</subject><subject>Apoptosis</subject><subject>Caspase 3 - metabolism</subject><subject>Caspase 7 - metabolism</subject><subject>caspases</subject><subject>Caspases - metabolism</subject><subject>Cell Membrane Permeability - physiology</subject><subject>Cellular Senescence - physiology</subject><subject>death</subject><subject>dephosphorylation</subject><subject>ejaculation</subject><subject>Enzyme Activation</subject><subject>Enzymes</subject><subject>flow cytometry</subject><subject>Flow Cytometry - veterinary</subject><subject>Horses</subject><subject>Horses - physiology</subject><subject>Hydrogen peroxide</subject><subject>Hydrogen Peroxide - metabolism</subject><subject>hydrogen production</subject><subject>incubation period</subject><subject>kinematics</subject><subject>Male</subject><subject>membrane permeability</subject><subject>membrane potential</subject><subject>Membrane Potential, Mitochondrial - physiology</subject><subject>mitochondrial membrane</subject><subject>Phosphorylation</subject><subject>Proto-Oncogene Proteins c-akt - metabolism</subject><subject>Semen Preservation - veterinary</subject><subject>Senescence</subject><subject>Sperm</subject><subject>sperm motility</subject><subject>Sperm Motility - physiology</subject><subject>spermatozoa</subject><subject>Spermatozoa - physiology</subject><subject>Spermatozoa - ultrastructure</subject><subject>stallions</subject><subject>superoxide anion</subject><subject>Time Factors</subject><issn>0936-6768</issn><issn>1439-0531</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2014</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNp1kl9v0zAUxS0EYmXwwBcAS7yARDY7dvznsWrZhjSxat3Eo-U4NyVbGmd2Auu3x122PSBh2bqS_TtH1z5G6D0lRzSN41DZI5ozzl6gGeVMZ6Rg9CWaEc1EJqRQB-hNjDeE0EJJ-Rod5FynKcUM3S1s7G0EPHdD89sOje--4rNdFfwGOryC4O-bCvAq-Gp0-1NsuwrPbwe8hP6Xj2mFXfugwxfOjQEvx9B0G7webNvud9c9hC1eQwfRQefgLXpV2zbCu8d6iK5Pvl0tzrLzi9Pvi_l55jhXLJPKlYzRgvOCaVIVBChVBbHKcWEl5wTqPF2VaSdLBYzmqgZRlUyJsqhdqdkh-jz59sHfjRAHs21SB21rO_BjNMlaFoxorRL66R_0xo-hS93tKaYkoYQn6stEueBjDFCbPjRbG3aGErPPwaQczEMOif3w6DiWW6ieyaeHT8DxBPxpWtj938lcLudPltmkaOIA988KG26NkEwW5uePUyNXKqeLkytzmfiPE19bb-wmNNFcr3NCefoGjOdCs7_KZ6kq</recordid><startdate>201408</startdate><enddate>201408</enddate><creator>Gallardo Bolaños, JM</creator><creator>Balao da Silva, C</creator><creator>Martín Muñoz, P</creator><creator>Plaza Dávila, M</creator><creator>Ezquerra, J</creator><creator>Aparicio, IM</creator><creator>Tapia, JA</creator><creator>Ortega Ferrusola, C</creator><creator>Peña, FJ</creator><general>Blackwell Science</general><general>Blackwell Publishing Ltd</general><scope>FBQ</scope><scope>BSCLL</scope><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>7QO</scope><scope>8FD</scope><scope>FR3</scope><scope>P64</scope><scope>7X8</scope></search><sort><creationdate>201408</creationdate><title>Caspase Activation, Hydrogen Peroxide Production and Akt Dephosphorylation Occur During Stallion Sperm Senescence</title><author>Gallardo Bolaños, JM ; Balao da Silva, C ; Martín Muñoz, P ; Plaza Dávila, M ; Ezquerra, J ; Aparicio, IM ; Tapia, JA ; Ortega Ferrusola, C ; Peña, FJ</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c4483-78cb3315445390d50e11850a8c46a7440ef243939c7b8e3128fe6db386b5fcb93</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2014</creationdate><topic>Animal reproduction</topic><topic>Animals</topic><topic>Apoptosis</topic><topic>Caspase 3 - metabolism</topic><topic>Caspase 7 - metabolism</topic><topic>caspases</topic><topic>Caspases - metabolism</topic><topic>Cell Membrane Permeability - physiology</topic><topic>Cellular Senescence - physiology</topic><topic>death</topic><topic>dephosphorylation</topic><topic>ejaculation</topic><topic>Enzyme Activation</topic><topic>Enzymes</topic><topic>flow cytometry</topic><topic>Flow Cytometry - veterinary</topic><topic>Horses</topic><topic>Horses - physiology</topic><topic>Hydrogen peroxide</topic><topic>Hydrogen Peroxide - metabolism</topic><topic>hydrogen production</topic><topic>incubation period</topic><topic>kinematics</topic><topic>Male</topic><topic>membrane permeability</topic><topic>membrane potential</topic><topic>Membrane Potential, Mitochondrial - physiology</topic><topic>mitochondrial membrane</topic><topic>Phosphorylation</topic><topic>Proto-Oncogene Proteins c-akt - metabolism</topic><topic>Semen Preservation - veterinary</topic><topic>Senescence</topic><topic>Sperm</topic><topic>sperm motility</topic><topic>Sperm Motility - physiology</topic><topic>spermatozoa</topic><topic>Spermatozoa - physiology</topic><topic>Spermatozoa - ultrastructure</topic><topic>stallions</topic><topic>superoxide anion</topic><topic>Time Factors</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Gallardo Bolaños, JM</creatorcontrib><creatorcontrib>Balao da Silva, C</creatorcontrib><creatorcontrib>Martín Muñoz, P</creatorcontrib><creatorcontrib>Plaza Dávila, M</creatorcontrib><creatorcontrib>Ezquerra, J</creatorcontrib><creatorcontrib>Aparicio, IM</creatorcontrib><creatorcontrib>Tapia, JA</creatorcontrib><creatorcontrib>Ortega Ferrusola, C</creatorcontrib><creatorcontrib>Peña, FJ</creatorcontrib><collection>AGRIS</collection><collection>Istex</collection><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>Biotechnology Research Abstracts</collection><collection>Technology Research Database</collection><collection>Engineering Research Database</collection><collection>Biotechnology and BioEngineering Abstracts</collection><collection>MEDLINE - Academic</collection><jtitle>Reproduction in domestic animals</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Gallardo Bolaños, JM</au><au>Balao da Silva, C</au><au>Martín Muñoz, P</au><au>Plaza Dávila, M</au><au>Ezquerra, J</au><au>Aparicio, IM</au><au>Tapia, JA</au><au>Ortega Ferrusola, C</au><au>Peña, FJ</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Caspase Activation, Hydrogen Peroxide Production and Akt Dephosphorylation Occur During Stallion Sperm Senescence</atitle><jtitle>Reproduction in domestic animals</jtitle><addtitle>Reprod Dom Anim</addtitle><date>2014-08</date><risdate>2014</risdate><volume>49</volume><issue>4</issue><spage>657</spage><epage>664</epage><pages>657-664</pages><issn>0936-6768</issn><eissn>1439-0531</eissn><abstract>To investigate the mechanisms inducing sperm death after ejaculation, stallion ejaculates were incubated in BWW media during 6 h at 37°C. At the beginning of the incubation period and after 1, 2, 4 and 6 h sperm motility and kinematics (CASA), mitochondrial membrane potential and membrane permeability and integrity were evaluated (flow cytometry). Also, at the same time intervals, active caspase 3, hydrogen peroxide, superoxide anion (flow cytometry) and Akt phosphorylation (flow cytometry) were evaluated. Major decreases in sperm function occurred after 6 h of incubation, although after 1 h decrease in the percentages of motile and progressive motile sperm occurred. The decrease observed in sperm functionality after 6 h of incubation was accompanied by a significant increase in the production of hydrogen peroxide and the greatest increase in caspase 3 activity. Additionally, the percentage of phosphorylated Akt reached a minimum after 6 h of incubation. These results provide evidences that sperm death during in vitro incubation is largely an apoptotic phenomena, probably stimulated by endogenous production of hydrogen peroxide and the lack of prosurvival factors maintaining Akt in a phosphorylated status. Disclosing molecular mechanisms leading to sperm death may help to develop new strategies for stallion sperm conservation.</abstract><cop>Germany</cop><pub>Blackwell Science</pub><pmid>24924976</pmid><doi>10.1111/rda.12343</doi><tpages>8</tpages></addata></record> |
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| subjects | Animal reproduction Animals Apoptosis Caspase 3 - metabolism Caspase 7 - metabolism caspases Caspases - metabolism Cell Membrane Permeability - physiology Cellular Senescence - physiology death dephosphorylation ejaculation Enzyme Activation Enzymes flow cytometry Flow Cytometry - veterinary Horses Horses - physiology Hydrogen peroxide Hydrogen Peroxide - metabolism hydrogen production incubation period kinematics Male membrane permeability membrane potential Membrane Potential, Mitochondrial - physiology mitochondrial membrane Phosphorylation Proto-Oncogene Proteins c-akt - metabolism Semen Preservation - veterinary Senescence Sperm sperm motility Sperm Motility - physiology spermatozoa Spermatozoa - physiology Spermatozoa - ultrastructure stallions superoxide anion Time Factors |
| title | Caspase Activation, Hydrogen Peroxide Production and Akt Dephosphorylation Occur During Stallion Sperm Senescence |
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