Chemically Assisted Enucleation Results in Higher G6PD Expression in Early Bovine Female Embryos Obtained by Somatic Cell Nuclear Transfer
Autor(a) principal: | |
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Data de Publicação: | 2012 |
Outros Autores: | , , , , , |
Tipo de documento: | Artigo |
Idioma: | eng |
Título da fonte: | Repositório Institucional da UNESP |
Texto Completo: | http://dx.doi.org/10.1089/cell.2011.0077 http://hdl.handle.net/11449/2548 |
Resumo: | Despite extensive efforts, low efficiency is still an issue in bovine somatic cell nuclear transfer (SCNT). The hypothesis of our study was that the use of cytoplasts produced by chemically assisted enucleation (EN) would improve nuclear reprogramming in nuclear transfer (NT)-derived embryos because it results in lower damage and higher cytoplasm content than conventional EN. For that purpose, we investigated the expression of two X-linked genes: X inactive-specific transcript (XIST) and glucose 6-phosphate dehydrogenase (G6PD). In the first experiment, gene expression was assessed in day-7 female blastocysts from embryonic cell NT (ECNT) groups [conventional, ECNT conv; chemically assisted, ECNT deme (demecolcine)]. Whereas in the ECNT conv group, only one embryo (25%; n = 4) expressed XIST transcripts, most embryos showed XIST expression (75%; n = 4) in the ECNT deme group. However, no significant differences in transcript abundance of XIST and G6PD were found when comparing the embryos from all groups. In a second experiment using somatic cells as nuclear donors, we evaluated gene expression profiles in female SCNT-derived embryos. No significant differences in relative abundance (RA) of XIST transcripts were observed among the groups. Nonetheless, higher (p < 0.05) levels of G6PD were observed in SCNT deme and in vitro-derived groups in comparison to SCNT conv. To know whether higher G6PD expression in embryos derived from SCNT chemically assisted EN indicates higher metabolism in embryos considered of superior quality or if the presence of higher reactive oxygen species (ROS) levels generated by the increased oxygen consumption triggers G6PD activation, the expression of genes related to stress response should be investigated in embryos produced by that technique. |
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Chemically Assisted Enucleation Results in Higher G6PD Expression in Early Bovine Female Embryos Obtained by Somatic Cell Nuclear TransferDespite extensive efforts, low efficiency is still an issue in bovine somatic cell nuclear transfer (SCNT). The hypothesis of our study was that the use of cytoplasts produced by chemically assisted enucleation (EN) would improve nuclear reprogramming in nuclear transfer (NT)-derived embryos because it results in lower damage and higher cytoplasm content than conventional EN. For that purpose, we investigated the expression of two X-linked genes: X inactive-specific transcript (XIST) and glucose 6-phosphate dehydrogenase (G6PD). In the first experiment, gene expression was assessed in day-7 female blastocysts from embryonic cell NT (ECNT) groups [conventional, ECNT conv; chemically assisted, ECNT deme (demecolcine)]. Whereas in the ECNT conv group, only one embryo (25%; n = 4) expressed XIST transcripts, most embryos showed XIST expression (75%; n = 4) in the ECNT deme group. However, no significant differences in transcript abundance of XIST and G6PD were found when comparing the embryos from all groups. In a second experiment using somatic cells as nuclear donors, we evaluated gene expression profiles in female SCNT-derived embryos. No significant differences in relative abundance (RA) of XIST transcripts were observed among the groups. Nonetheless, higher (p < 0.05) levels of G6PD were observed in SCNT deme and in vitro-derived groups in comparison to SCNT conv. To know whether higher G6PD expression in embryos derived from SCNT chemically assisted EN indicates higher metabolism in embryos considered of superior quality or if the presence of higher reactive oxygen species (ROS) levels generated by the increased oxygen consumption triggers G6PD activation, the expression of genes related to stress response should be investigated in embryos produced by that technique.Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Univ Estadual Paulista, Dept Med Vet Prevent & Reprod Anim, Jaboticabal, BrazilEmpresa Brasileira de Pesquisa Agropecuária (EMBRAPA) Dairy Cattle Res Ctr, Juiz de Fora, BrazilEmpresa Brasileira de Pesquisa Agropecuária (EMBRAPA) Cattle SE CPPSE, Sao Carlos, SP, BrazilUniv Estadual Paulista, Dept Med Vet Prevent & Reprod Anim, Jaboticabal, BrazilMary Ann Liebert, Inc.Universidade Estadual Paulista (Unesp)Empresa Brasileira de Pesquisa Agropecuária (EMBRAPA)Saraiva, Naiara Zoccal [UNESP]Oliveira, Clara Slade [UNESP]Drummond Tetzner, Tatiane Almeida [UNESP]de Lima, Marina Ragagnin [UNESP]de Melo, Danilas Salinet [UNESP]Meo Niciura, Simone CristinaGarcia, Joaquim Mansano [UNESP]2014-05-20T13:15:23Z2014-05-20T13:15:23Z2012-10-01info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/article425-435application/pdfhttp://dx.doi.org/10.1089/cell.2011.0077Cellular Reprogramming. New Rochelle: Mary Ann Liebert Inc., v. 14, n. 5, p. 425-435, 2012.2152-4971http://hdl.handle.net/11449/254810.1089/cell.2011.0077WOS:000309545700006WOS000309545700006.pdf2251116139872527Web of Sciencereponame:Repositório Institucional da UNESPinstname:Universidade Estadual Paulista (UNESP)instacron:UNESPengCellular Reprogramming1.4300,549info:eu-repo/semantics/openAccess2024-06-06T18:10:12Zoai:repositorio.unesp.br:11449/2548Repositório InstitucionalPUBhttp://repositorio.unesp.br/oai/requestopendoar:29462024-08-05T21:58:46.680955Repositório Institucional da UNESP - Universidade Estadual Paulista (UNESP)false |
dc.title.none.fl_str_mv |
Chemically Assisted Enucleation Results in Higher G6PD Expression in Early Bovine Female Embryos Obtained by Somatic Cell Nuclear Transfer |
title |
Chemically Assisted Enucleation Results in Higher G6PD Expression in Early Bovine Female Embryos Obtained by Somatic Cell Nuclear Transfer |
spellingShingle |
Chemically Assisted Enucleation Results in Higher G6PD Expression in Early Bovine Female Embryos Obtained by Somatic Cell Nuclear Transfer Saraiva, Naiara Zoccal [UNESP] |
title_short |
Chemically Assisted Enucleation Results in Higher G6PD Expression in Early Bovine Female Embryos Obtained by Somatic Cell Nuclear Transfer |
title_full |
Chemically Assisted Enucleation Results in Higher G6PD Expression in Early Bovine Female Embryos Obtained by Somatic Cell Nuclear Transfer |
title_fullStr |
Chemically Assisted Enucleation Results in Higher G6PD Expression in Early Bovine Female Embryos Obtained by Somatic Cell Nuclear Transfer |
title_full_unstemmed |
Chemically Assisted Enucleation Results in Higher G6PD Expression in Early Bovine Female Embryos Obtained by Somatic Cell Nuclear Transfer |
title_sort |
Chemically Assisted Enucleation Results in Higher G6PD Expression in Early Bovine Female Embryos Obtained by Somatic Cell Nuclear Transfer |
author |
Saraiva, Naiara Zoccal [UNESP] |
author_facet |
Saraiva, Naiara Zoccal [UNESP] Oliveira, Clara Slade [UNESP] Drummond Tetzner, Tatiane Almeida [UNESP] de Lima, Marina Ragagnin [UNESP] de Melo, Danilas Salinet [UNESP] Meo Niciura, Simone Cristina Garcia, Joaquim Mansano [UNESP] |
author_role |
author |
author2 |
Oliveira, Clara Slade [UNESP] Drummond Tetzner, Tatiane Almeida [UNESP] de Lima, Marina Ragagnin [UNESP] de Melo, Danilas Salinet [UNESP] Meo Niciura, Simone Cristina Garcia, Joaquim Mansano [UNESP] |
author2_role |
author author author author author author |
dc.contributor.none.fl_str_mv |
Universidade Estadual Paulista (Unesp) Empresa Brasileira de Pesquisa Agropecuária (EMBRAPA) |
dc.contributor.author.fl_str_mv |
Saraiva, Naiara Zoccal [UNESP] Oliveira, Clara Slade [UNESP] Drummond Tetzner, Tatiane Almeida [UNESP] de Lima, Marina Ragagnin [UNESP] de Melo, Danilas Salinet [UNESP] Meo Niciura, Simone Cristina Garcia, Joaquim Mansano [UNESP] |
description |
Despite extensive efforts, low efficiency is still an issue in bovine somatic cell nuclear transfer (SCNT). The hypothesis of our study was that the use of cytoplasts produced by chemically assisted enucleation (EN) would improve nuclear reprogramming in nuclear transfer (NT)-derived embryos because it results in lower damage and higher cytoplasm content than conventional EN. For that purpose, we investigated the expression of two X-linked genes: X inactive-specific transcript (XIST) and glucose 6-phosphate dehydrogenase (G6PD). In the first experiment, gene expression was assessed in day-7 female blastocysts from embryonic cell NT (ECNT) groups [conventional, ECNT conv; chemically assisted, ECNT deme (demecolcine)]. Whereas in the ECNT conv group, only one embryo (25%; n = 4) expressed XIST transcripts, most embryos showed XIST expression (75%; n = 4) in the ECNT deme group. However, no significant differences in transcript abundance of XIST and G6PD were found when comparing the embryos from all groups. In a second experiment using somatic cells as nuclear donors, we evaluated gene expression profiles in female SCNT-derived embryos. No significant differences in relative abundance (RA) of XIST transcripts were observed among the groups. Nonetheless, higher (p < 0.05) levels of G6PD were observed in SCNT deme and in vitro-derived groups in comparison to SCNT conv. To know whether higher G6PD expression in embryos derived from SCNT chemically assisted EN indicates higher metabolism in embryos considered of superior quality or if the presence of higher reactive oxygen species (ROS) levels generated by the increased oxygen consumption triggers G6PD activation, the expression of genes related to stress response should be investigated in embryos produced by that technique. |
publishDate |
2012 |
dc.date.none.fl_str_mv |
2012-10-01 2014-05-20T13:15:23Z 2014-05-20T13:15:23Z |
dc.type.status.fl_str_mv |
info:eu-repo/semantics/publishedVersion |
dc.type.driver.fl_str_mv |
info:eu-repo/semantics/article |
format |
article |
status_str |
publishedVersion |
dc.identifier.uri.fl_str_mv |
http://dx.doi.org/10.1089/cell.2011.0077 Cellular Reprogramming. New Rochelle: Mary Ann Liebert Inc., v. 14, n. 5, p. 425-435, 2012. 2152-4971 http://hdl.handle.net/11449/2548 10.1089/cell.2011.0077 WOS:000309545700006 WOS000309545700006.pdf 2251116139872527 |
url |
http://dx.doi.org/10.1089/cell.2011.0077 http://hdl.handle.net/11449/2548 |
identifier_str_mv |
Cellular Reprogramming. New Rochelle: Mary Ann Liebert Inc., v. 14, n. 5, p. 425-435, 2012. 2152-4971 10.1089/cell.2011.0077 WOS:000309545700006 WOS000309545700006.pdf 2251116139872527 |
dc.language.iso.fl_str_mv |
eng |
language |
eng |
dc.relation.none.fl_str_mv |
Cellular Reprogramming 1.430 0,549 |
dc.rights.driver.fl_str_mv |
info:eu-repo/semantics/openAccess |
eu_rights_str_mv |
openAccess |
dc.format.none.fl_str_mv |
425-435 application/pdf |
dc.publisher.none.fl_str_mv |
Mary Ann Liebert, Inc. |
publisher.none.fl_str_mv |
Mary Ann Liebert, Inc. |
dc.source.none.fl_str_mv |
Web of Science reponame:Repositório Institucional da UNESP instname:Universidade Estadual Paulista (UNESP) instacron:UNESP |
instname_str |
Universidade Estadual Paulista (UNESP) |
instacron_str |
UNESP |
institution |
UNESP |
reponame_str |
Repositório Institucional da UNESP |
collection |
Repositório Institucional da UNESP |
repository.name.fl_str_mv |
Repositório Institucional da UNESP - Universidade Estadual Paulista (UNESP) |
repository.mail.fl_str_mv |
|
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1808129380445061120 |