Embryo quality and production efficiency of porcine parthenotes is improved by phytohemagglutinin
In vitro production of porcine embryos has become routine in most laboratories but the yield and quality of the resultant blastocysts remain suboptimal. Phytohemagglutinin (PHA) is an N‐acetylgalactosamine/galactose sugar‐specific lectin with a wide variety of biological activities including mitogen...
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Veröffentlicht in: | Molecular reproduction and development 2007-04, Vol.74 (4), p.435-444 |
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description | In vitro production of porcine embryos has become routine in most laboratories but the yield and quality of the resultant blastocysts remain suboptimal. Phytohemagglutinin (PHA) is an N‐acetylgalactosamine/galactose sugar‐specific lectin with a wide variety of biological activities including mitogenesis, mediation of cell recognition, and agglutination of cells. This study was therefore, designed to investigate the effect of PHA on the preimplantation embryo development and quality of in vitro produced porcine parthenotes. Parthenogenetic presumptive diploid zygotes were produced in vitro by electrical activation and cultured in the absence or presence of PHA at different concentrations (0, 5, 10, 15, 20 µg/ml). There were no significant differences in the cleavage rate of porcine parthenotes in control and treatment groups at all tested concentrations of PHA (P 0.05). TUNEL labeling revealed that blastocysts in PHA group were less predisposed to biochemical apoptosis than in control group while total apoptosis and nuclear fragmentation remained unaltered. Real‐time reverse transcription‐polymerase chain reaction (RT‐PCR) analysis further revealed that PHA decreased the expression ratio of BAX/BCL‐XL and enhanced the relative abundance of IGF2 transcripts. Therefore, our study suggests that PHA improves the blastocyst yield and quality by enhancing blastocyst expansion, hatching, and total cell number and decreasing the apoptosis by positively modulating the expression of embryo survival related genes. Mol. Reprod. Dev. 74: 435–444, 2007. © 2006 Wiley‐Liss, Inc. |
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Phytohemagglutinin (PHA) is an N‐acetylgalactosamine/galactose sugar‐specific lectin with a wide variety of biological activities including mitogenesis, mediation of cell recognition, and agglutination of cells. This study was therefore, designed to investigate the effect of PHA on the preimplantation embryo development and quality of in vitro produced porcine parthenotes. Parthenogenetic presumptive diploid zygotes were produced in vitro by electrical activation and cultured in the absence or presence of PHA at different concentrations (0, 5, 10, 15, 20 µg/ml). There were no significant differences in the cleavage rate of porcine parthenotes in control and treatment groups at all tested concentrations of PHA (P < 0.05). However, supplementation of PHA at the concentration of 15 µg/ml significantly improved the blastocyst rate (68.9 ± 1.5% vs. 43.1 ± 4.1%), hatching rate (25.8 ± 3.1% vs. 8.9 ± 2.0%), and total nuclei number (95.5 ± 9.3 vs. 63.4 ± 4.3) when compared to control group (P > 0.05). TUNEL labeling revealed that blastocysts in PHA group were less predisposed to biochemical apoptosis than in control group while total apoptosis and nuclear fragmentation remained unaltered. Real‐time reverse transcription‐polymerase chain reaction (RT‐PCR) analysis further revealed that PHA decreased the expression ratio of BAX/BCL‐XL and enhanced the relative abundance of IGF2 transcripts. Therefore, our study suggests that PHA improves the blastocyst yield and quality by enhancing blastocyst expansion, hatching, and total cell number and decreasing the apoptosis by positively modulating the expression of embryo survival related genes. Mol. Reprod. Dev. 74: 435–444, 2007. © 2006 Wiley‐Liss, Inc.</description><identifier>ISSN: 1040-452X</identifier><identifier>EISSN: 1098-2795</identifier><identifier>DOI: 10.1002/mrd.20547</identifier><identifier>PMID: 16998814</identifier><identifier>CODEN: MREDEE</identifier><language>eng</language><publisher>Hoboken: Wiley Subscription Services, Inc., A Wiley Company</publisher><subject>Animals ; apoptosis ; Apoptosis - drug effects ; bcl-2-Associated X Protein - metabolism ; bcl-X Protein - metabolism ; Biological and medical sciences ; Cell Survival - drug effects ; Cloning, Organism - methods ; development rate ; Dose-Response Relationship, Drug ; Embryo Culture Techniques ; Embryo, Mammalian - cytology ; Embryo, Mammalian - drug effects ; Embryo, Mammalian - metabolism ; Embryology: invertebrates and vertebrates. Teratology ; Embryonic Development - drug effects ; Female ; Fundamental and applied biological sciences. Psychology ; gene expression ; Insulin-Like Growth Factor II - metabolism ; Molecular embryology ; Parthenogenesis ; phytohemagglutinin ; Phytohemagglutinins - pharmacology ; pig embryo ; Random Allocation ; Swine</subject><ispartof>Molecular reproduction and development, 2007-04, Vol.74 (4), p.435-444</ispartof><rights>Copyright © 2006 Wiley‐Liss, Inc.</rights><rights>2007 INIST-CNRS</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c4577-212e02e0f32bcba58fc98443e9ed955672abe3558b801f8663e21d8f3a434f883</citedby><cites>FETCH-LOGICAL-c4577-212e02e0f32bcba58fc98443e9ed955672abe3558b801f8663e21d8f3a434f883</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://onlinelibrary.wiley.com/doi/pdf/10.1002%2Fmrd.20547$$EPDF$$P50$$Gwiley$$H</linktopdf><linktohtml>$$Uhttps://onlinelibrary.wiley.com/doi/full/10.1002%2Fmrd.20547$$EHTML$$P50$$Gwiley$$H</linktohtml><link.rule.ids>314,776,780,1411,27901,27902,45550,45551</link.rule.ids><backlink>$$Uhttp://pascal-francis.inist.fr/vibad/index.php?action=getRecordDetail&idt=18543845$$DView record in Pascal Francis$$Hfree_for_read</backlink><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/16998814$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Gupta, Mukesh Kumar</creatorcontrib><creatorcontrib>Uhm, Sang Jun</creatorcontrib><creatorcontrib>Han, Dong Wook</creatorcontrib><creatorcontrib>Lee, Hoon Taek</creatorcontrib><title>Embryo quality and production efficiency of porcine parthenotes is improved by phytohemagglutinin</title><title>Molecular reproduction and development</title><addtitle>Mol. Reprod. Dev</addtitle><description>In vitro production of porcine embryos has become routine in most laboratories but the yield and quality of the resultant blastocysts remain suboptimal. Phytohemagglutinin (PHA) is an N‐acetylgalactosamine/galactose sugar‐specific lectin with a wide variety of biological activities including mitogenesis, mediation of cell recognition, and agglutination of cells. This study was therefore, designed to investigate the effect of PHA on the preimplantation embryo development and quality of in vitro produced porcine parthenotes. Parthenogenetic presumptive diploid zygotes were produced in vitro by electrical activation and cultured in the absence or presence of PHA at different concentrations (0, 5, 10, 15, 20 µg/ml). There were no significant differences in the cleavage rate of porcine parthenotes in control and treatment groups at all tested concentrations of PHA (P < 0.05). However, supplementation of PHA at the concentration of 15 µg/ml significantly improved the blastocyst rate (68.9 ± 1.5% vs. 43.1 ± 4.1%), hatching rate (25.8 ± 3.1% vs. 8.9 ± 2.0%), and total nuclei number (95.5 ± 9.3 vs. 63.4 ± 4.3) when compared to control group (P > 0.05). TUNEL labeling revealed that blastocysts in PHA group were less predisposed to biochemical apoptosis than in control group while total apoptosis and nuclear fragmentation remained unaltered. Real‐time reverse transcription‐polymerase chain reaction (RT‐PCR) analysis further revealed that PHA decreased the expression ratio of BAX/BCL‐XL and enhanced the relative abundance of IGF2 transcripts. Therefore, our study suggests that PHA improves the blastocyst yield and quality by enhancing blastocyst expansion, hatching, and total cell number and decreasing the apoptosis by positively modulating the expression of embryo survival related genes. Mol. Reprod. Dev. 74: 435–444, 2007. © 2006 Wiley‐Liss, Inc.</description><subject>Animals</subject><subject>apoptosis</subject><subject>Apoptosis - drug effects</subject><subject>bcl-2-Associated X Protein - metabolism</subject><subject>bcl-X Protein - metabolism</subject><subject>Biological and medical sciences</subject><subject>Cell Survival - drug effects</subject><subject>Cloning, Organism - methods</subject><subject>development rate</subject><subject>Dose-Response Relationship, Drug</subject><subject>Embryo Culture Techniques</subject><subject>Embryo, Mammalian - cytology</subject><subject>Embryo, Mammalian - drug effects</subject><subject>Embryo, Mammalian - metabolism</subject><subject>Embryology: invertebrates and vertebrates. Teratology</subject><subject>Embryonic Development - drug effects</subject><subject>Female</subject><subject>Fundamental and applied biological sciences. Psychology</subject><subject>gene expression</subject><subject>Insulin-Like Growth Factor II - metabolism</subject><subject>Molecular embryology</subject><subject>Parthenogenesis</subject><subject>phytohemagglutinin</subject><subject>Phytohemagglutinins - pharmacology</subject><subject>pig embryo</subject><subject>Random Allocation</subject><subject>Swine</subject><issn>1040-452X</issn><issn>1098-2795</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2007</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNp1kEtv1TAQhS0EoqWw4A8gb0BikdbP2FnS0gfSBQQCys5ynHGvIYlTOynk3ze395aukEaas_jOnNFB6CUlh5QQdtSl5pARKdQjtE9JpQumKvl4owUphGQ_99CznH8RQqpKk6doj5aL0FTsI3va1WmO-HqybRhnbPsGDyk2kxtD7DF4H1yA3s04ejzE5EIPeLBpXEMfR8g4LNMtjhtocD3jYT2PcQ2dvbpqpzH0oX-OnnjbZnix2wfo-9npt5OLYvX5_MPJu1XhhFSqYJQBWcZzVrvaSu1dpYXgUEFTSVkqZmvgUupaE-p1WXJgtNGeW8GF15ofoDfbu8sz1xPk0XQhO2hb20Ocsil1VVJSbsC3W9ClmHMCb4YUOptmQ4nZ9GmWPs1dnwv7and0qjtoHshdgQvwegfY7Gzrk-1dyA-cloJrIRfuaMv9CS3M_080H7--v48uto6QR_j7z2HTb1MqrqS5_HRu2PGXlVQ_lLnkt3xmnOk</recordid><startdate>200704</startdate><enddate>200704</enddate><creator>Gupta, Mukesh Kumar</creator><creator>Uhm, Sang Jun</creator><creator>Han, Dong Wook</creator><creator>Lee, Hoon Taek</creator><general>Wiley Subscription Services, Inc., A Wiley Company</general><general>Wiley-Liss</general><scope>BSCLL</scope><scope>IQODW</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>7X8</scope></search><sort><creationdate>200704</creationdate><title>Embryo quality and production efficiency of porcine parthenotes is improved by phytohemagglutinin</title><author>Gupta, Mukesh Kumar ; Uhm, Sang Jun ; Han, Dong Wook ; Lee, Hoon Taek</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c4577-212e02e0f32bcba58fc98443e9ed955672abe3558b801f8663e21d8f3a434f883</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2007</creationdate><topic>Animals</topic><topic>apoptosis</topic><topic>Apoptosis - drug effects</topic><topic>bcl-2-Associated X Protein - metabolism</topic><topic>bcl-X Protein - metabolism</topic><topic>Biological and medical sciences</topic><topic>Cell Survival - drug effects</topic><topic>Cloning, Organism - methods</topic><topic>development rate</topic><topic>Dose-Response Relationship, Drug</topic><topic>Embryo Culture Techniques</topic><topic>Embryo, Mammalian - cytology</topic><topic>Embryo, Mammalian - drug effects</topic><topic>Embryo, Mammalian - metabolism</topic><topic>Embryology: invertebrates and vertebrates. Teratology</topic><topic>Embryonic Development - drug effects</topic><topic>Female</topic><topic>Fundamental and applied biological sciences. Psychology</topic><topic>gene expression</topic><topic>Insulin-Like Growth Factor II - metabolism</topic><topic>Molecular embryology</topic><topic>Parthenogenesis</topic><topic>phytohemagglutinin</topic><topic>Phytohemagglutinins - pharmacology</topic><topic>pig embryo</topic><topic>Random Allocation</topic><topic>Swine</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Gupta, Mukesh Kumar</creatorcontrib><creatorcontrib>Uhm, Sang Jun</creatorcontrib><creatorcontrib>Han, Dong Wook</creatorcontrib><creatorcontrib>Lee, Hoon Taek</creatorcontrib><collection>Istex</collection><collection>Pascal-Francis</collection><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>Molecular reproduction and development</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Gupta, Mukesh Kumar</au><au>Uhm, Sang Jun</au><au>Han, Dong Wook</au><au>Lee, Hoon Taek</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Embryo quality and production efficiency of porcine parthenotes is improved by phytohemagglutinin</atitle><jtitle>Molecular reproduction and development</jtitle><addtitle>Mol. Reprod. Dev</addtitle><date>2007-04</date><risdate>2007</risdate><volume>74</volume><issue>4</issue><spage>435</spage><epage>444</epage><pages>435-444</pages><issn>1040-452X</issn><eissn>1098-2795</eissn><coden>MREDEE</coden><abstract>In vitro production of porcine embryos has become routine in most laboratories but the yield and quality of the resultant blastocysts remain suboptimal. Phytohemagglutinin (PHA) is an N‐acetylgalactosamine/galactose sugar‐specific lectin with a wide variety of biological activities including mitogenesis, mediation of cell recognition, and agglutination of cells. This study was therefore, designed to investigate the effect of PHA on the preimplantation embryo development and quality of in vitro produced porcine parthenotes. Parthenogenetic presumptive diploid zygotes were produced in vitro by electrical activation and cultured in the absence or presence of PHA at different concentrations (0, 5, 10, 15, 20 µg/ml). There were no significant differences in the cleavage rate of porcine parthenotes in control and treatment groups at all tested concentrations of PHA (P < 0.05). However, supplementation of PHA at the concentration of 15 µg/ml significantly improved the blastocyst rate (68.9 ± 1.5% vs. 43.1 ± 4.1%), hatching rate (25.8 ± 3.1% vs. 8.9 ± 2.0%), and total nuclei number (95.5 ± 9.3 vs. 63.4 ± 4.3) when compared to control group (P > 0.05). TUNEL labeling revealed that blastocysts in PHA group were less predisposed to biochemical apoptosis than in control group while total apoptosis and nuclear fragmentation remained unaltered. Real‐time reverse transcription‐polymerase chain reaction (RT‐PCR) analysis further revealed that PHA decreased the expression ratio of BAX/BCL‐XL and enhanced the relative abundance of IGF2 transcripts. Therefore, our study suggests that PHA improves the blastocyst yield and quality by enhancing blastocyst expansion, hatching, and total cell number and decreasing the apoptosis by positively modulating the expression of embryo survival related genes. Mol. Reprod. Dev. 74: 435–444, 2007. © 2006 Wiley‐Liss, Inc.</abstract><cop>Hoboken</cop><pub>Wiley Subscription Services, Inc., A Wiley Company</pub><pmid>16998814</pmid><doi>10.1002/mrd.20547</doi><tpages>10</tpages></addata></record> |
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subjects | Animals apoptosis Apoptosis - drug effects bcl-2-Associated X Protein - metabolism bcl-X Protein - metabolism Biological and medical sciences Cell Survival - drug effects Cloning, Organism - methods development rate Dose-Response Relationship, Drug Embryo Culture Techniques Embryo, Mammalian - cytology Embryo, Mammalian - drug effects Embryo, Mammalian - metabolism Embryology: invertebrates and vertebrates. Teratology Embryonic Development - drug effects Female Fundamental and applied biological sciences. Psychology gene expression Insulin-Like Growth Factor II - metabolism Molecular embryology Parthenogenesis phytohemagglutinin Phytohemagglutinins - pharmacology pig embryo Random Allocation Swine |
title | Embryo quality and production efficiency of porcine parthenotes is improved by phytohemagglutinin |
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