Improved preimplantation development of bovine ICSI embryos generated with spermatozoa pretreated with membrane-destabilizing agents lysolecithin and Triton X-100
In cattle, intracytoplasmic sperm injection (ICSI) has a low efficiency. The acrosome content may be responsible for this effect because of the large amount of hydrolytic enzymes that are released within the oocyte. With the aim of removing the acrosome and destabilize the membranes, cryopreserved b...
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| Veröffentlicht in: | Theriogenology 2016-10, Vol.86 (6), p.1489-1497 |
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| creator | Zambrano, Fabiola Aguila, Luis Arias, María E. Sánchez, Raúl Felmer, Ricardo |
| description | In cattle, intracytoplasmic sperm injection (ICSI) has a low efficiency. The acrosome content may be responsible for this effect because of the large amount of hydrolytic enzymes that are released within the oocyte. With the aim of removing the acrosome and destabilize the membranes, cryopreserved bovine spermatozoa were treated with lysolecithin (LL) and Triton X-100 (TX) at different concentrations. We evaluated the membrane integrity, the acrosome integrity, DNA integrity, and the variation of phospholipase C zeta. The rates of development (cleavage and blastocysts) were also evaluated along with pronuclear formation and the embryo quality. Spermatozoa incubated with LL and TX (0.01%, 0.02%, 0.03%, and 0.04%) decreased (P |
| doi_str_mv | 10.1016/j.theriogenology.2016.05.007 |
| format | Article |
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The acrosome content may be responsible for this effect because of the large amount of hydrolytic enzymes that are released within the oocyte. With the aim of removing the acrosome and destabilize the membranes, cryopreserved bovine spermatozoa were treated with lysolecithin (LL) and Triton X-100 (TX) at different concentrations. We evaluated the membrane integrity, the acrosome integrity, DNA integrity, and the variation of phospholipase C zeta. The rates of development (cleavage and blastocysts) were also evaluated along with pronuclear formation and the embryo quality. Spermatozoa incubated with LL and TX (0.01%, 0.02%, 0.03%, and 0.04%) decreased (P < 0.0001) sperm viability in a dose-dependent manner. The acrosome reaction was also increased (P < 0.0001) in all tested concentrations of LL and TX achieving 100% at 0.05% concentration in both treatments. Terminal deoxynucleotidyl transferase dUTP nick-end labeling assay reported an increase (P < 0.05) in DNA fragmentation only with the highest concentration of LL (0.06%), whereas all concentrations assessed of TX reported an increased respect to the control. Phospholipase C zeta expression decreased (P < 0.05) in spermatozoa treated with LL and TX at all concentrations tested. A higher cleavage rate was observed in ICSI-TX (66%) and ICSI-LL (65%) groups compared with the untreated control group (51%) and the blastocyst formation rate significantly increased in the ICSI-LL group (29%) compared with the control (21%). No differences were observed in the pronuclear formation and quality of the embryos. In conclusion, the destabilization of the plasma membrane and the release of the acrosomal content with LL and TX before ICSI improve the rate of embryonic development, without affecting the quality of the embryos produced by this technique.</description><identifier>ISSN: 0093-691X</identifier><identifier>EISSN: 1879-3231</identifier><identifier>DOI: 10.1016/j.theriogenology.2016.05.007</identifier><identifier>PMID: 27325573</identifier><language>eng</language><publisher>United States: Elsevier Inc</publisher><subject>Acrosome ; Acrosome - drug effects ; Acrosome - physiology ; Acrosome Reaction - drug effects ; Animals ; Bovine spermatozoa ; Cattle - embryology ; Cell Membrane - drug effects ; Cryopreservation - veterinary ; DNA Fragmentation - drug effects ; Embryonic Development ; Female ; Glycoproteins - pharmacology ; ICSI ; In Situ Nick-End Labeling ; Lysophospholipase - pharmacology ; Male ; Membrane disruption ; Octoxynol - pharmacology ; Semen Preservation - veterinary ; Sperm Injections, Intracytoplasmic - veterinary ; Spermatozoa - drug effects ; Spermatozoa - physiology ; Spermatozoa - ultrastructure ; Type C Phospholipases - analysis ; Type C Phospholipases - metabolism</subject><ispartof>Theriogenology, 2016-10, Vol.86 (6), p.1489-1497</ispartof><rights>2016 Elsevier Inc.</rights><rights>Copyright © 2016 Elsevier Inc. All rights reserved.</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c386t-cd5e9027ff67539ada414f77342db69c0bea377456191ebe7f7134d468489b003</citedby><cites>FETCH-LOGICAL-c386t-cd5e9027ff67539ada414f77342db69c0bea377456191ebe7f7134d468489b003</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktohtml>$$Uhttps://dx.doi.org/10.1016/j.theriogenology.2016.05.007$$EHTML$$P50$$Gelsevier$$H</linktohtml><link.rule.ids>314,778,782,3539,27913,27914,45984</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/27325573$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Zambrano, Fabiola</creatorcontrib><creatorcontrib>Aguila, Luis</creatorcontrib><creatorcontrib>Arias, María E.</creatorcontrib><creatorcontrib>Sánchez, Raúl</creatorcontrib><creatorcontrib>Felmer, Ricardo</creatorcontrib><title>Improved preimplantation development of bovine ICSI embryos generated with spermatozoa pretreated with membrane-destabilizing agents lysolecithin and Triton X-100</title><title>Theriogenology</title><addtitle>Theriogenology</addtitle><description>In cattle, intracytoplasmic sperm injection (ICSI) has a low efficiency. The acrosome content may be responsible for this effect because of the large amount of hydrolytic enzymes that are released within the oocyte. With the aim of removing the acrosome and destabilize the membranes, cryopreserved bovine spermatozoa were treated with lysolecithin (LL) and Triton X-100 (TX) at different concentrations. We evaluated the membrane integrity, the acrosome integrity, DNA integrity, and the variation of phospholipase C zeta. The rates of development (cleavage and blastocysts) were also evaluated along with pronuclear formation and the embryo quality. Spermatozoa incubated with LL and TX (0.01%, 0.02%, 0.03%, and 0.04%) decreased (P < 0.0001) sperm viability in a dose-dependent manner. The acrosome reaction was also increased (P < 0.0001) in all tested concentrations of LL and TX achieving 100% at 0.05% concentration in both treatments. Terminal deoxynucleotidyl transferase dUTP nick-end labeling assay reported an increase (P < 0.05) in DNA fragmentation only with the highest concentration of LL (0.06%), whereas all concentrations assessed of TX reported an increased respect to the control. Phospholipase C zeta expression decreased (P < 0.05) in spermatozoa treated with LL and TX at all concentrations tested. A higher cleavage rate was observed in ICSI-TX (66%) and ICSI-LL (65%) groups compared with the untreated control group (51%) and the blastocyst formation rate significantly increased in the ICSI-LL group (29%) compared with the control (21%). No differences were observed in the pronuclear formation and quality of the embryos. In conclusion, the destabilization of the plasma membrane and the release of the acrosomal content with LL and TX before ICSI improve the rate of embryonic development, without affecting the quality of the embryos produced by this technique.</description><subject>Acrosome</subject><subject>Acrosome - drug effects</subject><subject>Acrosome - physiology</subject><subject>Acrosome Reaction - drug effects</subject><subject>Animals</subject><subject>Bovine spermatozoa</subject><subject>Cattle - embryology</subject><subject>Cell Membrane - drug effects</subject><subject>Cryopreservation - veterinary</subject><subject>DNA Fragmentation - drug effects</subject><subject>Embryonic Development</subject><subject>Female</subject><subject>Glycoproteins - pharmacology</subject><subject>ICSI</subject><subject>In Situ Nick-End Labeling</subject><subject>Lysophospholipase - pharmacology</subject><subject>Male</subject><subject>Membrane disruption</subject><subject>Octoxynol - pharmacology</subject><subject>Semen Preservation - veterinary</subject><subject>Sperm Injections, Intracytoplasmic - veterinary</subject><subject>Spermatozoa - drug effects</subject><subject>Spermatozoa - physiology</subject><subject>Spermatozoa - ultrastructure</subject><subject>Type C Phospholipases - analysis</subject><subject>Type C Phospholipases - metabolism</subject><issn>0093-691X</issn><issn>1879-3231</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2016</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqNUcFu1DAUtBCIbgu_gHzgwCXBjhM7kbigFaUrVeJAkXqzHPtl61VsB9u7aPs5fClebQFx4_Qkv5l54xmE3lJSU0L5-12dHyDasAUf5rA91k15rUlXEyKeoRXtxVCxhtHnaEXIwCo-0PsLdJnSjhDCOKcv0UUjWNN1gq3Qz41bYjiAwUsE65ZZ-ayyDR4bOMAcFgc-4zDhMRysB7xZf91gcGM8hoSLBYgqF_IPmx9wWiA6lcNjUCe1HOHvzp04ykNlIGU12tk-Wr_FqkjkhOdjCjPoArQeK2_wXbS5eLivKCGv0ItJzQleP80r9O360936prr98nmz_nhbadbzXGnTwUAaMU1cdGxQRrW0nYRgbWNGPmgygmJCtB2nA4URxCQoa03L-7YfxhLNFXp31i2BfN8Xm9LZpGEukUDYJ0l72rRsIFQU6IczVMeQUoRJLtE6FY-SEnlqSe7kvy3JU0uSdLK0VOhvni7tRwfmD_l3LQVwfQZA-e_BQpRJW_AajI2gszTB_t-lXweIsMo</recordid><startdate>20161001</startdate><enddate>20161001</enddate><creator>Zambrano, Fabiola</creator><creator>Aguila, Luis</creator><creator>Arias, María E.</creator><creator>Sánchez, Raúl</creator><creator>Felmer, Ricardo</creator><general>Elsevier Inc</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>20161001</creationdate><title>Improved preimplantation development of bovine ICSI embryos generated with spermatozoa pretreated with membrane-destabilizing agents lysolecithin and Triton X-100</title><author>Zambrano, Fabiola ; Aguila, Luis ; Arias, María E. ; Sánchez, Raúl ; Felmer, Ricardo</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c386t-cd5e9027ff67539ada414f77342db69c0bea377456191ebe7f7134d468489b003</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2016</creationdate><topic>Acrosome</topic><topic>Acrosome - drug effects</topic><topic>Acrosome - physiology</topic><topic>Acrosome Reaction - drug effects</topic><topic>Animals</topic><topic>Bovine spermatozoa</topic><topic>Cattle - embryology</topic><topic>Cell Membrane - drug effects</topic><topic>Cryopreservation - veterinary</topic><topic>DNA Fragmentation - drug effects</topic><topic>Embryonic Development</topic><topic>Female</topic><topic>Glycoproteins - pharmacology</topic><topic>ICSI</topic><topic>In Situ Nick-End Labeling</topic><topic>Lysophospholipase - pharmacology</topic><topic>Male</topic><topic>Membrane disruption</topic><topic>Octoxynol - pharmacology</topic><topic>Semen Preservation - veterinary</topic><topic>Sperm Injections, Intracytoplasmic - veterinary</topic><topic>Spermatozoa - drug effects</topic><topic>Spermatozoa - physiology</topic><topic>Spermatozoa - ultrastructure</topic><topic>Type C Phospholipases - analysis</topic><topic>Type C Phospholipases - metabolism</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Zambrano, Fabiola</creatorcontrib><creatorcontrib>Aguila, Luis</creatorcontrib><creatorcontrib>Arias, María E.</creatorcontrib><creatorcontrib>Sánchez, Raúl</creatorcontrib><creatorcontrib>Felmer, Ricardo</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>Theriogenology</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Zambrano, Fabiola</au><au>Aguila, Luis</au><au>Arias, María E.</au><au>Sánchez, Raúl</au><au>Felmer, Ricardo</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Improved preimplantation development of bovine ICSI embryos generated with spermatozoa pretreated with membrane-destabilizing agents lysolecithin and Triton X-100</atitle><jtitle>Theriogenology</jtitle><addtitle>Theriogenology</addtitle><date>2016-10-01</date><risdate>2016</risdate><volume>86</volume><issue>6</issue><spage>1489</spage><epage>1497</epage><pages>1489-1497</pages><issn>0093-691X</issn><eissn>1879-3231</eissn><abstract>In cattle, intracytoplasmic sperm injection (ICSI) has a low efficiency. The acrosome content may be responsible for this effect because of the large amount of hydrolytic enzymes that are released within the oocyte. With the aim of removing the acrosome and destabilize the membranes, cryopreserved bovine spermatozoa were treated with lysolecithin (LL) and Triton X-100 (TX) at different concentrations. We evaluated the membrane integrity, the acrosome integrity, DNA integrity, and the variation of phospholipase C zeta. The rates of development (cleavage and blastocysts) were also evaluated along with pronuclear formation and the embryo quality. Spermatozoa incubated with LL and TX (0.01%, 0.02%, 0.03%, and 0.04%) decreased (P < 0.0001) sperm viability in a dose-dependent manner. The acrosome reaction was also increased (P < 0.0001) in all tested concentrations of LL and TX achieving 100% at 0.05% concentration in both treatments. Terminal deoxynucleotidyl transferase dUTP nick-end labeling assay reported an increase (P < 0.05) in DNA fragmentation only with the highest concentration of LL (0.06%), whereas all concentrations assessed of TX reported an increased respect to the control. Phospholipase C zeta expression decreased (P < 0.05) in spermatozoa treated with LL and TX at all concentrations tested. A higher cleavage rate was observed in ICSI-TX (66%) and ICSI-LL (65%) groups compared with the untreated control group (51%) and the blastocyst formation rate significantly increased in the ICSI-LL group (29%) compared with the control (21%). No differences were observed in the pronuclear formation and quality of the embryos. In conclusion, the destabilization of the plasma membrane and the release of the acrosomal content with LL and TX before ICSI improve the rate of embryonic development, without affecting the quality of the embryos produced by this technique.</abstract><cop>United States</cop><pub>Elsevier Inc</pub><pmid>27325573</pmid><doi>10.1016/j.theriogenology.2016.05.007</doi><tpages>9</tpages></addata></record> |
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| subjects | Acrosome Acrosome - drug effects Acrosome - physiology Acrosome Reaction - drug effects Animals Bovine spermatozoa Cattle - embryology Cell Membrane - drug effects Cryopreservation - veterinary DNA Fragmentation - drug effects Embryonic Development Female Glycoproteins - pharmacology ICSI In Situ Nick-End Labeling Lysophospholipase - pharmacology Male Membrane disruption Octoxynol - pharmacology Semen Preservation - veterinary Sperm Injections, Intracytoplasmic - veterinary Spermatozoa - drug effects Spermatozoa - physiology Spermatozoa - ultrastructure Type C Phospholipases - analysis Type C Phospholipases - metabolism |
| title | Improved preimplantation development of bovine ICSI embryos generated with spermatozoa pretreated with membrane-destabilizing agents lysolecithin and Triton X-100 |
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