Cholesterol-loaded-cyclodextrins improve the post-thaw quality of stallion sperm
An unacceptable proportion of stallion sperm do not survive the freeze-thaw process. The hypothesis of this study was that adding cholesterol to a stallion semen extender would stabilise the sperm membrane, resulting in an improved post-thaw semen quality in terms of increased sperm viability, membr...
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| Veröffentlicht in: | Animal reproduction science 2014-03, Vol.145 (3-4), p.123-129 |
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| creator | Murphy, C. English, A.M. Holden, S.A. Fair, S. |
| description | An unacceptable proportion of stallion sperm do not survive the freeze-thaw process. The hypothesis of this study was that adding cholesterol to a stallion semen extender would stabilise the sperm membrane, resulting in an improved post-thaw semen quality in terms of increased sperm viability, membrane integrity and fluidity, and reduced oxidative stress. Semen was collected from three stallions and diluted in four extenders: TALP; TALP+0.75mg methyl-β-cyclodextrin–cholesterol (MβCD)/mL (MβCD0.75); TALP+1.5mg MβCD-cholesterol/mL (MβCD1.5); and Equipro. Following 15min incubation, samples were centrifuged and diluted to 100×106sperm/mL, frozen in 0.5mL straws and stored in liquid nitrogen. Sperm from each treatment was assessed for progressive linear motility (PLM) and acceptable membrane integrity under hypotonic conditions on a phase contrast microscope at 1000× while viability, membrane fluidity and superoxide generation were assessed by flow cytometry. The MβCD1.5 and MβCD0.75 treatments had a greater proportion of viable sperm than the TALP treatment (P |
| doi_str_mv | 10.1016/j.anireprosci.2014.01.013 |
| format | Article |
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The hypothesis of this study was that adding cholesterol to a stallion semen extender would stabilise the sperm membrane, resulting in an improved post-thaw semen quality in terms of increased sperm viability, membrane integrity and fluidity, and reduced oxidative stress. Semen was collected from three stallions and diluted in four extenders: TALP; TALP+0.75mg methyl-β-cyclodextrin–cholesterol (MβCD)/mL (MβCD0.75); TALP+1.5mg MβCD-cholesterol/mL (MβCD1.5); and Equipro. Following 15min incubation, samples were centrifuged and diluted to 100×106sperm/mL, frozen in 0.5mL straws and stored in liquid nitrogen. Sperm from each treatment was assessed for progressive linear motility (PLM) and acceptable membrane integrity under hypotonic conditions on a phase contrast microscope at 1000× while viability, membrane fluidity and superoxide generation were assessed by flow cytometry. The MβCD1.5 and MβCD0.75 treatments had a greater proportion of viable sperm than the TALP treatment (P<0.01). There was no effect of treatment on PLM or membrane integrity. The MβCD1.5 treatment had a greater proportion of viable sperm positive for membrane fluidity than the TALP treatment (P<0.05). The MβCD1.5 and MβCD0.75 treatments had a lesser proportion of viable sperm positive for superoxide generation than the TALP treatment (P<0.001). This study has demonstrated that adding cholesterol to stallion sperm prior to cryopreservation increases post-thaw viability, with these viable sperm being of better quality in terms of increased membrane fluidity and reduced superoxide generation.</description><identifier>ISSN: 0378-4320</identifier><identifier>EISSN: 1873-2232</identifier><identifier>DOI: 10.1016/j.anireprosci.2014.01.013</identifier><identifier>PMID: 24583046</identifier><language>eng</language><publisher>Netherlands: Elsevier B.V</publisher><subject>Animals ; Cholesterol ; Cholesterol - chemistry ; Cholesterol - pharmacology ; Cryoprotective Agents - chemistry ; Cryoprotective Agents - pharmacology ; Cyclodextrins - chemistry ; Cyclodextrins - pharmacology ; Equine ; Horses - physiology ; Male ; Membrane fluidity ; Reactive oxygen species ; Semen Analysis ; Semen Preservation - methods ; Semen Preservation - veterinary ; Sperm ; Spermatozoa - drug effects ; Viability</subject><ispartof>Animal reproduction science, 2014-03, Vol.145 (3-4), p.123-129</ispartof><rights>2014 Elsevier B.V.</rights><rights>Copyright © 2014 Elsevier B.V. All rights reserved.</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c377t-4fd127aa26327a0614b08639b9f5fcacb96a73f018976949cf4d341230a2a2583</citedby><cites>FETCH-LOGICAL-c377t-4fd127aa26327a0614b08639b9f5fcacb96a73f018976949cf4d341230a2a2583</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktohtml>$$Uhttps://dx.doi.org/10.1016/j.anireprosci.2014.01.013$$EHTML$$P50$$Gelsevier$$H</linktohtml><link.rule.ids>314,780,784,3550,27924,27925,45995</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/24583046$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Murphy, C.</creatorcontrib><creatorcontrib>English, A.M.</creatorcontrib><creatorcontrib>Holden, S.A.</creatorcontrib><creatorcontrib>Fair, S.</creatorcontrib><title>Cholesterol-loaded-cyclodextrins improve the post-thaw quality of stallion sperm</title><title>Animal reproduction science</title><addtitle>Anim Reprod Sci</addtitle><description>An unacceptable proportion of stallion sperm do not survive the freeze-thaw process. The hypothesis of this study was that adding cholesterol to a stallion semen extender would stabilise the sperm membrane, resulting in an improved post-thaw semen quality in terms of increased sperm viability, membrane integrity and fluidity, and reduced oxidative stress. Semen was collected from three stallions and diluted in four extenders: TALP; TALP+0.75mg methyl-β-cyclodextrin–cholesterol (MβCD)/mL (MβCD0.75); TALP+1.5mg MβCD-cholesterol/mL (MβCD1.5); and Equipro. Following 15min incubation, samples were centrifuged and diluted to 100×106sperm/mL, frozen in 0.5mL straws and stored in liquid nitrogen. Sperm from each treatment was assessed for progressive linear motility (PLM) and acceptable membrane integrity under hypotonic conditions on a phase contrast microscope at 1000× while viability, membrane fluidity and superoxide generation were assessed by flow cytometry. The MβCD1.5 and MβCD0.75 treatments had a greater proportion of viable sperm than the TALP treatment (P<0.01). There was no effect of treatment on PLM or membrane integrity. The MβCD1.5 treatment had a greater proportion of viable sperm positive for membrane fluidity than the TALP treatment (P<0.05). The MβCD1.5 and MβCD0.75 treatments had a lesser proportion of viable sperm positive for superoxide generation than the TALP treatment (P<0.001). This study has demonstrated that adding cholesterol to stallion sperm prior to cryopreservation increases post-thaw viability, with these viable sperm being of better quality in terms of increased membrane fluidity and reduced superoxide generation.</description><subject>Animals</subject><subject>Cholesterol</subject><subject>Cholesterol - chemistry</subject><subject>Cholesterol - pharmacology</subject><subject>Cryoprotective Agents - chemistry</subject><subject>Cryoprotective Agents - pharmacology</subject><subject>Cyclodextrins - chemistry</subject><subject>Cyclodextrins - pharmacology</subject><subject>Equine</subject><subject>Horses - physiology</subject><subject>Male</subject><subject>Membrane fluidity</subject><subject>Reactive oxygen species</subject><subject>Semen Analysis</subject><subject>Semen Preservation - methods</subject><subject>Semen Preservation - veterinary</subject><subject>Sperm</subject><subject>Spermatozoa - drug effects</subject><subject>Viability</subject><issn>0378-4320</issn><issn>1873-2232</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2014</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqNkM1LAzEQxYMotlb_BVlvXrZOPprdPUrxCwQ96Dmk2Vmakt3UJFX735tSFY_CwFzevDfvR8gFhSkFKq9WUz3YgOvgo7FTBlRMgebhB2RM64qXjHF2SMbAq7oUnMGInMS4AoBKyuaYjJiY1RyEHJPn-dI7jAmDd6XzusW2NFvjfIufKdghFrbPOe9YpCUWax9TmZb6o3jbaGfTtvBdEZN2zvqhiGsM_Sk56rSLePa9J-T19uZlfl8-Pt09zK8fS8OrKpWiaymrtGaS5wWSigXUkjeLppt1RptFI3XFO6B1U8lGNKYTLReUcdBMs_z9hFzuffN3b5vcQPU2GnROD-g3UdEZ1ILKXDlLm73UZGAxYKfWwfY6bBUFtQOqVuoPULUDqoDm4fn2_Dtms-ix_b38IZgF870Ac9l3i0FlCxwMttnQJNV6-4-YL4nkjcw</recordid><startdate>20140301</startdate><enddate>20140301</enddate><creator>Murphy, C.</creator><creator>English, A.M.</creator><creator>Holden, S.A.</creator><creator>Fair, S.</creator><general>Elsevier B.V</general><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>7X8</scope></search><sort><creationdate>20140301</creationdate><title>Cholesterol-loaded-cyclodextrins improve the post-thaw quality of stallion sperm</title><author>Murphy, C. ; English, A.M. ; Holden, S.A. ; Fair, S.</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c377t-4fd127aa26327a0614b08639b9f5fcacb96a73f018976949cf4d341230a2a2583</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2014</creationdate><topic>Animals</topic><topic>Cholesterol</topic><topic>Cholesterol - chemistry</topic><topic>Cholesterol - pharmacology</topic><topic>Cryoprotective Agents - chemistry</topic><topic>Cryoprotective Agents - pharmacology</topic><topic>Cyclodextrins - chemistry</topic><topic>Cyclodextrins - pharmacology</topic><topic>Equine</topic><topic>Horses - physiology</topic><topic>Male</topic><topic>Membrane fluidity</topic><topic>Reactive oxygen species</topic><topic>Semen Analysis</topic><topic>Semen Preservation - methods</topic><topic>Semen Preservation - veterinary</topic><topic>Sperm</topic><topic>Spermatozoa - drug effects</topic><topic>Viability</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Murphy, C.</creatorcontrib><creatorcontrib>English, A.M.</creatorcontrib><creatorcontrib>Holden, S.A.</creatorcontrib><creatorcontrib>Fair, S.</creatorcontrib><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>MEDLINE - Academic</collection><jtitle>Animal reproduction science</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Murphy, C.</au><au>English, A.M.</au><au>Holden, S.A.</au><au>Fair, S.</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Cholesterol-loaded-cyclodextrins improve the post-thaw quality of stallion sperm</atitle><jtitle>Animal reproduction science</jtitle><addtitle>Anim Reprod Sci</addtitle><date>2014-03-01</date><risdate>2014</risdate><volume>145</volume><issue>3-4</issue><spage>123</spage><epage>129</epage><pages>123-129</pages><issn>0378-4320</issn><eissn>1873-2232</eissn><abstract>An unacceptable proportion of stallion sperm do not survive the freeze-thaw process. The hypothesis of this study was that adding cholesterol to a stallion semen extender would stabilise the sperm membrane, resulting in an improved post-thaw semen quality in terms of increased sperm viability, membrane integrity and fluidity, and reduced oxidative stress. Semen was collected from three stallions and diluted in four extenders: TALP; TALP+0.75mg methyl-β-cyclodextrin–cholesterol (MβCD)/mL (MβCD0.75); TALP+1.5mg MβCD-cholesterol/mL (MβCD1.5); and Equipro. Following 15min incubation, samples were centrifuged and diluted to 100×106sperm/mL, frozen in 0.5mL straws and stored in liquid nitrogen. Sperm from each treatment was assessed for progressive linear motility (PLM) and acceptable membrane integrity under hypotonic conditions on a phase contrast microscope at 1000× while viability, membrane fluidity and superoxide generation were assessed by flow cytometry. The MβCD1.5 and MβCD0.75 treatments had a greater proportion of viable sperm than the TALP treatment (P<0.01). There was no effect of treatment on PLM or membrane integrity. The MβCD1.5 treatment had a greater proportion of viable sperm positive for membrane fluidity than the TALP treatment (P<0.05). The MβCD1.5 and MβCD0.75 treatments had a lesser proportion of viable sperm positive for superoxide generation than the TALP treatment (P<0.001). This study has demonstrated that adding cholesterol to stallion sperm prior to cryopreservation increases post-thaw viability, with these viable sperm being of better quality in terms of increased membrane fluidity and reduced superoxide generation.</abstract><cop>Netherlands</cop><pub>Elsevier B.V</pub><pmid>24583046</pmid><doi>10.1016/j.anireprosci.2014.01.013</doi><tpages>7</tpages></addata></record> |
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| subjects | Animals Cholesterol Cholesterol - chemistry Cholesterol - pharmacology Cryoprotective Agents - chemistry Cryoprotective Agents - pharmacology Cyclodextrins - chemistry Cyclodextrins - pharmacology Equine Horses - physiology Male Membrane fluidity Reactive oxygen species Semen Analysis Semen Preservation - methods Semen Preservation - veterinary Sperm Spermatozoa - drug effects Viability |
| title | Cholesterol-loaded-cyclodextrins improve the post-thaw quality of stallion sperm |
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