Effects of discontinuous Percoll gradient centrifugation on the quality of bovine spermatozoa evaluated with computer-assisted semen analysis and fluorescent probes association
Summary The objective of this study was to evaluate the quality of bovine frozen‐thawed sperm cells after Percoll gradient centrifugation. Frozen semen doses were obtained from six bulls of different breeds, including three taurine and three Zebu animals. Four ejaculates per bull were evaluated befo...
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| Veröffentlicht in: | Andrologia 2012-02, Vol.44 (1), p.9-15 |
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| creator | Oliveira, L. Z. Arruda, R. P. Celeghini, E. C. C. de Andrade, A. F. C. Perini, A. P. Resende, M. V. Miguel, M. C. V. Lucio, A. C. Hossepian de Lima, V. F. M. |
| description | Summary
The objective of this study was to evaluate the quality of bovine frozen‐thawed sperm cells after Percoll gradient centrifugation. Frozen semen doses were obtained from six bulls of different breeds, including three taurine and three Zebu animals. Four ejaculates per bull were evaluated before and after discontinuous Percoll gradient centrifugation. Sperm motility was assessed by computer‐assisted semen analysis and the integrity of the plasma and acrosomal membranes, as well as mitochondrial function, were evaluated using a combination of fluorescent probes propidium iodide, fluorescein isothiocyanate‐conjugated Pisum sativum agglutinin and 5,5′,6,6′‐tetrachloro‐1,1′,3,3′‐tetraethylbenzimidazolcarbocyanine iodide. The procedure of Percoll gradient centrifugation increased the percentage of total and progressive sperm motility, beat frequency, rectilinear motility, linearity and rapidly moving cells. In addition, the percentage of cells with intact plasma membrane and mitochondrial membrane potential was increased in post‐centrifugation samples. However, the percentage of sperm cells with intact acrosomal membrane was markedly reduced. The method used selected the motile cells with intact plasma membrane and higher mitochondrial functionality in frozen‐thawed bull semen, but processing, centrifugation and/or the Percoll medium caused damage to the acrosomal membrane. |
| doi_str_mv | 10.1111/j.1439-0272.2010.01096.x |
| format | Article |
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The objective of this study was to evaluate the quality of bovine frozen‐thawed sperm cells after Percoll gradient centrifugation. Frozen semen doses were obtained from six bulls of different breeds, including three taurine and three Zebu animals. Four ejaculates per bull were evaluated before and after discontinuous Percoll gradient centrifugation. Sperm motility was assessed by computer‐assisted semen analysis and the integrity of the plasma and acrosomal membranes, as well as mitochondrial function, were evaluated using a combination of fluorescent probes propidium iodide, fluorescein isothiocyanate‐conjugated Pisum sativum agglutinin and 5,5′,6,6′‐tetrachloro‐1,1′,3,3′‐tetraethylbenzimidazolcarbocyanine iodide. The procedure of Percoll gradient centrifugation increased the percentage of total and progressive sperm motility, beat frequency, rectilinear motility, linearity and rapidly moving cells. In addition, the percentage of cells with intact plasma membrane and mitochondrial membrane potential was increased in post‐centrifugation samples. However, the percentage of sperm cells with intact acrosomal membrane was markedly reduced. The method used selected the motile cells with intact plasma membrane and higher mitochondrial functionality in frozen‐thawed bull semen, but processing, centrifugation and/or the Percoll medium caused damage to the acrosomal membrane.</description><identifier>ISSN: 0303-4569</identifier><identifier>EISSN: 1439-0272</identifier><identifier>DOI: 10.1111/j.1439-0272.2010.01096.x</identifier><identifier>PMID: 21615453</identifier><language>eng</language><publisher>Oxford, UK: Blackwell Publishing Ltd</publisher><subject>Acrosome integrity ; Animals ; Cattle ; Cell Separation - veterinary ; Centrifugation, Density Gradient ; Computers ; Cryopreservation - veterinary ; cryopreserved semen ; Fluorescent Dyes ; Male ; Povidone ; Semen Analysis - veterinary ; Semen Preservation - veterinary ; Silicon Dioxide ; Sperm Motility ; sperm selection ; Spermatozoa - physiology</subject><ispartof>Andrologia, 2012-02, Vol.44 (1), p.9-15</ispartof><rights>2011 Blackwell Verlag GmbH</rights><rights>2011 Blackwell Verlag GmbH.</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c4076-8c7c550cdb7bc19ca0039695072895cccef1a6b0e19a16a913196ee30b7c8e93</citedby></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://onlinelibrary.wiley.com/doi/pdf/10.1111%2Fj.1439-0272.2010.01096.x$$EPDF$$P50$$Gwiley$$H</linktopdf><linktohtml>$$Uhttps://onlinelibrary.wiley.com/doi/full/10.1111%2Fj.1439-0272.2010.01096.x$$EHTML$$P50$$Gwiley$$H</linktohtml><link.rule.ids>314,776,780,1411,27901,27902,45550,45551</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/21615453$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Oliveira, L. Z.</creatorcontrib><creatorcontrib>Arruda, R. P.</creatorcontrib><creatorcontrib>Celeghini, E. C. C.</creatorcontrib><creatorcontrib>de Andrade, A. F. C.</creatorcontrib><creatorcontrib>Perini, A. P.</creatorcontrib><creatorcontrib>Resende, M. V.</creatorcontrib><creatorcontrib>Miguel, M. C. V.</creatorcontrib><creatorcontrib>Lucio, A. C.</creatorcontrib><creatorcontrib>Hossepian de Lima, V. F. M.</creatorcontrib><title>Effects of discontinuous Percoll gradient centrifugation on the quality of bovine spermatozoa evaluated with computer-assisted semen analysis and fluorescent probes association</title><title>Andrologia</title><addtitle>Andrologia</addtitle><description>Summary
The objective of this study was to evaluate the quality of bovine frozen‐thawed sperm cells after Percoll gradient centrifugation. Frozen semen doses were obtained from six bulls of different breeds, including three taurine and three Zebu animals. Four ejaculates per bull were evaluated before and after discontinuous Percoll gradient centrifugation. Sperm motility was assessed by computer‐assisted semen analysis and the integrity of the plasma and acrosomal membranes, as well as mitochondrial function, were evaluated using a combination of fluorescent probes propidium iodide, fluorescein isothiocyanate‐conjugated Pisum sativum agglutinin and 5,5′,6,6′‐tetrachloro‐1,1′,3,3′‐tetraethylbenzimidazolcarbocyanine iodide. The procedure of Percoll gradient centrifugation increased the percentage of total and progressive sperm motility, beat frequency, rectilinear motility, linearity and rapidly moving cells. In addition, the percentage of cells with intact plasma membrane and mitochondrial membrane potential was increased in post‐centrifugation samples. However, the percentage of sperm cells with intact acrosomal membrane was markedly reduced. The method used selected the motile cells with intact plasma membrane and higher mitochondrial functionality in frozen‐thawed bull semen, but processing, centrifugation and/or the Percoll medium caused damage to the acrosomal membrane.</description><subject>Acrosome integrity</subject><subject>Animals</subject><subject>Cattle</subject><subject>Cell Separation - veterinary</subject><subject>Centrifugation, Density Gradient</subject><subject>Computers</subject><subject>Cryopreservation - veterinary</subject><subject>cryopreserved semen</subject><subject>Fluorescent Dyes</subject><subject>Male</subject><subject>Povidone</subject><subject>Semen Analysis - veterinary</subject><subject>Semen Preservation - veterinary</subject><subject>Silicon Dioxide</subject><subject>Sperm Motility</subject><subject>sperm selection</subject><subject>Spermatozoa - physiology</subject><issn>0303-4569</issn><issn>1439-0272</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2012</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNo9UU1v3CAQRVWrJkrzFypuPXkDxsBy6CHKZ6VV2kOk9oYwHidssdkATnb7q_oTi7PpjvgYvZnHoPcQwpQsaImz9YI2TFWklvWiJgUtW4nF9h06PhTeo2PCCKsaLtQROk1pTUo0XMqm-YiOaioobzg7Rn-v-h5sTjj0uHPJhjG7cQpTwj8g2uA9foimczBmbMsRXT89mOzCiMvKj4CfJuNd3s38Njy7EXDaQBxMDn-CwfBs_GQydPjF5Udsw7CZMsTKpOTSDCcYYMRmNH5XkJJ0uPdTiJDmcXgTQwsFTilY9zr3E_rQG5_g9O0-QffXV_cXt9Xq-823i_NVZRsiRbW00nJObNfK1lJlDSFMCcWJrJeKW2uhp0a0BKgyVBhFGVUCgJFW2iUodoK-7J8tP3iaIGU9FHXAezNCUUcr2ggheV2Xzs9vnVM7QKc30Q0m7vR_jUvD133Di_OwO9Qp0bOdeq1n1_Tsmp7t1K926q0-v7ucs8Kv9vxZse2Bb-JvLSSTXP-8u9GX4tfqWhKib9k_0lmnjA</recordid><startdate>201202</startdate><enddate>201202</enddate><creator>Oliveira, L. Z.</creator><creator>Arruda, R. P.</creator><creator>Celeghini, E. C. C.</creator><creator>de Andrade, A. F. C.</creator><creator>Perini, A. P.</creator><creator>Resende, M. V.</creator><creator>Miguel, M. C. V.</creator><creator>Lucio, A. C.</creator><creator>Hossepian de Lima, V. F. M.</creator><general>Blackwell Publishing Ltd</general><scope>BSCLL</scope><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>7X8</scope></search><sort><creationdate>201202</creationdate><title>Effects of discontinuous Percoll gradient centrifugation on the quality of bovine spermatozoa evaluated with computer-assisted semen analysis and fluorescent probes association</title><author>Oliveira, L. Z. ; Arruda, R. P. ; Celeghini, E. C. C. ; de Andrade, A. F. C. ; Perini, A. P. ; Resende, M. V. ; Miguel, M. C. V. ; Lucio, A. C. ; Hossepian de Lima, V. F. M.</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c4076-8c7c550cdb7bc19ca0039695072895cccef1a6b0e19a16a913196ee30b7c8e93</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2012</creationdate><topic>Acrosome integrity</topic><topic>Animals</topic><topic>Cattle</topic><topic>Cell Separation - veterinary</topic><topic>Centrifugation, Density Gradient</topic><topic>Computers</topic><topic>Cryopreservation - veterinary</topic><topic>cryopreserved semen</topic><topic>Fluorescent Dyes</topic><topic>Male</topic><topic>Povidone</topic><topic>Semen Analysis - veterinary</topic><topic>Semen Preservation - veterinary</topic><topic>Silicon Dioxide</topic><topic>Sperm Motility</topic><topic>sperm selection</topic><topic>Spermatozoa - physiology</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Oliveira, L. Z.</creatorcontrib><creatorcontrib>Arruda, R. P.</creatorcontrib><creatorcontrib>Celeghini, E. C. C.</creatorcontrib><creatorcontrib>de Andrade, A. F. C.</creatorcontrib><creatorcontrib>Perini, A. P.</creatorcontrib><creatorcontrib>Resende, M. V.</creatorcontrib><creatorcontrib>Miguel, M. C. V.</creatorcontrib><creatorcontrib>Lucio, A. C.</creatorcontrib><creatorcontrib>Hossepian de Lima, V. F. M.</creatorcontrib><collection>Istex</collection><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>MEDLINE - Academic</collection><jtitle>Andrologia</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Oliveira, L. Z.</au><au>Arruda, R. P.</au><au>Celeghini, E. C. C.</au><au>de Andrade, A. F. C.</au><au>Perini, A. P.</au><au>Resende, M. V.</au><au>Miguel, M. C. V.</au><au>Lucio, A. C.</au><au>Hossepian de Lima, V. F. M.</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Effects of discontinuous Percoll gradient centrifugation on the quality of bovine spermatozoa evaluated with computer-assisted semen analysis and fluorescent probes association</atitle><jtitle>Andrologia</jtitle><addtitle>Andrologia</addtitle><date>2012-02</date><risdate>2012</risdate><volume>44</volume><issue>1</issue><spage>9</spage><epage>15</epage><pages>9-15</pages><issn>0303-4569</issn><eissn>1439-0272</eissn><abstract>Summary
The objective of this study was to evaluate the quality of bovine frozen‐thawed sperm cells after Percoll gradient centrifugation. Frozen semen doses were obtained from six bulls of different breeds, including three taurine and three Zebu animals. Four ejaculates per bull were evaluated before and after discontinuous Percoll gradient centrifugation. Sperm motility was assessed by computer‐assisted semen analysis and the integrity of the plasma and acrosomal membranes, as well as mitochondrial function, were evaluated using a combination of fluorescent probes propidium iodide, fluorescein isothiocyanate‐conjugated Pisum sativum agglutinin and 5,5′,6,6′‐tetrachloro‐1,1′,3,3′‐tetraethylbenzimidazolcarbocyanine iodide. The procedure of Percoll gradient centrifugation increased the percentage of total and progressive sperm motility, beat frequency, rectilinear motility, linearity and rapidly moving cells. In addition, the percentage of cells with intact plasma membrane and mitochondrial membrane potential was increased in post‐centrifugation samples. However, the percentage of sperm cells with intact acrosomal membrane was markedly reduced. The method used selected the motile cells with intact plasma membrane and higher mitochondrial functionality in frozen‐thawed bull semen, but processing, centrifugation and/or the Percoll medium caused damage to the acrosomal membrane.</abstract><cop>Oxford, UK</cop><pub>Blackwell Publishing Ltd</pub><pmid>21615453</pmid><doi>10.1111/j.1439-0272.2010.01096.x</doi><tpages>7</tpages><oa>free_for_read</oa></addata></record> |
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| source | MEDLINE; Wiley Online Library Journals Frontfile Complete |
| subjects | Acrosome integrity Animals Cattle Cell Separation - veterinary Centrifugation, Density Gradient Computers Cryopreservation - veterinary cryopreserved semen Fluorescent Dyes Male Povidone Semen Analysis - veterinary Semen Preservation - veterinary Silicon Dioxide Sperm Motility sperm selection Spermatozoa - physiology |
| title | Effects of discontinuous Percoll gradient centrifugation on the quality of bovine spermatozoa evaluated with computer-assisted semen analysis and fluorescent probes association |
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