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
Hauptverfasser: Gupta, Mukesh Kumar, Uhm, Sang Jun, Han, Dong Wook, Lee, Hoon Taek
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Han, Dong Wook
Lee, Hoon Taek
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.
doi_str_mv 10.1002/mrd.20547
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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 &lt; 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 &gt; 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. 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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 &lt; 0.05). 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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 &lt; 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 &gt; 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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source MEDLINE; Wiley Online Library Journals Frontfile Complete
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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