Influência do FSH sobre os aspectos celulares e moleculares na maturação in vitro de complexos cumulus-ovócitos de bovinos

Detalhes bibliográficos
Autor(a) principal: CANTANHÊDE, Ludymila Furtado
Data de Publicação: 2018
Tipo de documento: Tese
Idioma: por
Título da fonte: Biblioteca Digital de Teses e Dissertações da UFRPE
Texto Completo: http://www.tede2.ufrpe.br:8080/tede2/handle/tede2/7507
Resumo: Biotechniques applied to bovine reproduction still have limitations and many are linked to a lack of knowledge about an update of macromolecules added to the medium of in vitro oocyte maturation (IVM), embryonic development process and cell difference, as well as genes of inhibitors related to pluripotency in in vitro production in this species. Follicle stimulating hormone (FSH) is present in most IVM protocols because of its importance in large cumulus cells, estradiol synthesis and cytoplasmic and nuclear maturation of the oocyte. However, few studies report an influence of FSH on gene activity during maturation. Considering the identification and modification of the biochemistry of the medium and mechanisms of action of the genes contributed to improvements in the various biotechniques applied to the reproduction to increase the production of animals of high genetic value, this work had the objective of evaluating an influence of FSH in cumulus during maturation of bovine oocytes. Hormonal and biochemical analyzes of the in vitro maturation medium before and MAB, with and FSH (G-FSH, G-FSHIN, G-CTL, G-CTLIN), as well as an analysis of the expression of pluripotency related genes in cumulus of cattle matured with and without FSH (G-FSH and G-CTL) collected in slaughterhouses of the Zona da Mata and Agreste of Pernambuco. A concentration of estradiol was observed by immunoassay and concentration of protein, glycogen, sugar and lipid (biochemical analysis) through a spectrophotometer in the analysis of real-time PCR (RT-qPCR). For statistical analysis, normality was assessed by Shapiro-Wilk test to be submitted to analysis of variance (Test F) and, or Tukey HSD test, level of significance of 5.0%. Increased cumulus cell amplitudes were observed during maturation in the FSH medium when compared to the non-FSH medium. In addition, FSH influences glycogen consumption by cells, decreasing their concentration not medium, due to the need for high energy demand for the process. Influence was observed in the expression of RONIN and CMYC, in which the hormone stimulates RONIN activity and represses CMYC. Thus, FSH influences a maturation of oocytes, besides also acting in the energetic activity of the cumulus-oocyte complex of bovine without cellular and molecular physics.
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spelling OLIVEIRA, Marcos Antonio Lemos deMOURA, Marcelo TigreMOURA, Marcelo TigreBARTOLOMEU, Cláudio CoutinhoTEIXEIRA, Álvaro Aguiar Coelhohttp://lattes.cnpq.br/7950707554487640CANTANHÊDE, Ludymila Furtado2018-09-04T12:56:30Z2018-02-23CANTANHÊDE, Ludymila Furtado. Influência do FSH sobre os aspectos celulares e moleculares na maturação in vitro de complexos cumulus-ovócitos de bovinos. 2018. 114 f. Tese (Programa de Pós-Graduação em Ciência Veterinária) - Universidade Federal Rural de Pernambuco, Recife.http://www.tede2.ufrpe.br:8080/tede2/handle/tede2/7507Biotechniques applied to bovine reproduction still have limitations and many are linked to a lack of knowledge about an update of macromolecules added to the medium of in vitro oocyte maturation (IVM), embryonic development process and cell difference, as well as genes of inhibitors related to pluripotency in in vitro production in this species. Follicle stimulating hormone (FSH) is present in most IVM protocols because of its importance in large cumulus cells, estradiol synthesis and cytoplasmic and nuclear maturation of the oocyte. However, few studies report an influence of FSH on gene activity during maturation. Considering the identification and modification of the biochemistry of the medium and mechanisms of action of the genes contributed to improvements in the various biotechniques applied to the reproduction to increase the production of animals of high genetic value, this work had the objective of evaluating an influence of FSH in cumulus during maturation of bovine oocytes. Hormonal and biochemical analyzes of the in vitro maturation medium before and MAB, with and FSH (G-FSH, G-FSHIN, G-CTL, G-CTLIN), as well as an analysis of the expression of pluripotency related genes in cumulus of cattle matured with and without FSH (G-FSH and G-CTL) collected in slaughterhouses of the Zona da Mata and Agreste of Pernambuco. A concentration of estradiol was observed by immunoassay and concentration of protein, glycogen, sugar and lipid (biochemical analysis) through a spectrophotometer in the analysis of real-time PCR (RT-qPCR). For statistical analysis, normality was assessed by Shapiro-Wilk test to be submitted to analysis of variance (Test F) and, or Tukey HSD test, level of significance of 5.0%. Increased cumulus cell amplitudes were observed during maturation in the FSH medium when compared to the non-FSH medium. In addition, FSH influences glycogen consumption by cells, decreasing their concentration not medium, due to the need for high energy demand for the process. Influence was observed in the expression of RONIN and CMYC, in which the hormone stimulates RONIN activity and represses CMYC. Thus, FSH influences a maturation of oocytes, besides also acting in the energetic activity of the cumulus-oocyte complex of bovine without cellular and molecular physics.As biotécnicas aplicadas à reprodução de bovinos ainda possuem limitações e muitas estão ligadas a falta de conhecimento sobre a atuação de macromoléculas adicionadas ao meio de maturação ovocitária in vitro (MIV), ao processo de desenvolvimento embrionário e diferenciação celular, bem como as alterações de genes relacionados à pluripotência na produção in vitro nesta espécie. O hormônio folículo estimulante (FSH) está presente na maioria dos protocolos de MIV, devido a sua importância na expansão das células do cumulus, síntese de estradiol e maturação citoplasmática e nuclear do ovócito. Porém, poucos estudos relatam a influência do FSH na atividade gênica durante a maturação. Visto que a identificação e elucidação das modificações bioquímicas do meio e dos mecanismos de ação desses genes contribuiriam para melhorias nas diversas biotécnicas aplicadas à reprodução e para o aumento da produção de animais de alto valor genético, este trabalho teve como objetivo avaliar a influencia do FSH em células do cumulus durante a maturação de ovócitos bovinos. Foram feitas análises hormonal e bioquímicas do meio de maturação in vitro antes e após a MIV, com e sem FSH (G-FSH, G-FSHIN, G-CTL, G-CTLIN) bem como a análise da expressão de genes relacionados à pluripotência em células do cumulus de bovinos maturados com e sem FSH (G-FSH e G-CTL) coletados em abatedouros da região da Zona da Mata e Agreste de Pernambuco. Foi observada a concentração de estradiol por imunoensaio e concentração de proteína, glicogênio, açúcar e lipídeo (análise bioquímica) através de espectofotômetro na análise de expressão relativa por PCR em tempo real (RT-qPCR). Para análise estatística avaliou-se a normalidade pelo teste de Shapiro-Wilk para serem então submetidos à análise de variância (ANOVA), Teste F e, posteriormente o teste de Tukey HSD, nível de significância de 5,0%. Foi observada maior expansão das células do cumulus durante a maturação em meio contendo FSH, quando comparado ao meio sem FSH. Além disso, o FSH influenciou o consumo de glicogênio pelas células, diminuindo sua concentração no meio, sugerindo grande demanda de energia para o processo. Observou-se influência do FSH sobre a expressão de RONIN e CMYC, nos quais o hormônio estimula a atividade de RONIN e reprime CMYC. Dessa forma, o FSH influencia a maturação de ovócitos bovinos, além de atuar também na atividade energética do complexo cumulus-ovócito de nos níveis celulares e moleculares.Submitted by Mario BC (mario@bc.ufrpe.br) on 2018-09-04T12:56:30Z No. of bitstreams: 1 Ludymila Furtado Cantanhede.pdf: 2024123 bytes, checksum: c6c240c3b28427794e299888dcab2f37 (MD5)Made available in DSpace on 2018-09-04T12:56:30Z (GMT). No. of bitstreams: 1 Ludymila Furtado Cantanhede.pdf: 2024123 bytes, checksum: c6c240c3b28427794e299888dcab2f37 (MD5) Previous issue date: 2018-02-23Conselho Nacional de Pesquisa e Desenvolvimento Científico e Tecnológico - CNPqapplication/pdfporUniversidade Federal Rural de PernambucoPrograma de Pós-Graduação em Ciência VeterináriaUFRPEBrasilDepartamento de Medicina VeterináriaMaturação ovocitáriaHormônio folículo estimulanteBovinoCIENCIAS AGRARIAS::MEDICINA VETERINARIAInfluência do FSH sobre os aspectos celulares e moleculares na maturação in vitro de complexos cumulus-ovócitos de bovinosinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/doctoralThesis-3061482854177903105600600600600-3020210563763616780453670264235017319-2555911436985713659info:eu-repo/semantics/openAccessreponame:Biblioteca Digital de Teses e Dissertações da UFRPEinstname:Universidade Federal Rural de Pernambuco (UFRPE)instacron:UFRPEORIGINALLudymila Furtado Cantanhede.pdfLudymila Furtado Cantanhede.pdfapplication/pdf2024123http://www.tede2.ufrpe.br:8080/tede2/bitstream/tede2/7507/2/Ludymila+Furtado+Cantanhede.pdfc6c240c3b28427794e299888dcab2f37MD52LICENSElicense.txtlicense.txttext/plain; charset=utf-82165http://www.tede2.ufrpe.br:8080/tede2/bitstream/tede2/7507/1/license.txtbd3efa91386c1718a7f26a329fdcb468MD51tede2/75072018-09-04 09:56:30.9oai:tede2: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Biblioteca Digital de Teses e Dissertaçõeshttp://www.tede2.ufrpe.br:8080/tede/PUBhttp://www.tede2.ufrpe.br:8080/oai/requestbdtd@ufrpe.br ||bdtd@ufrpe.bropendoar:2018-09-04T12:56:30Biblioteca Digital de Teses e Dissertações da UFRPE - Universidade Federal Rural de Pernambuco (UFRPE)false
dc.title.por.fl_str_mv Influência do FSH sobre os aspectos celulares e moleculares na maturação in vitro de complexos cumulus-ovócitos de bovinos
title Influência do FSH sobre os aspectos celulares e moleculares na maturação in vitro de complexos cumulus-ovócitos de bovinos
spellingShingle Influência do FSH sobre os aspectos celulares e moleculares na maturação in vitro de complexos cumulus-ovócitos de bovinos
CANTANHÊDE, Ludymila Furtado
Maturação ovocitária
Hormônio folículo estimulante
Bovino
CIENCIAS AGRARIAS::MEDICINA VETERINARIA
title_short Influência do FSH sobre os aspectos celulares e moleculares na maturação in vitro de complexos cumulus-ovócitos de bovinos
title_full Influência do FSH sobre os aspectos celulares e moleculares na maturação in vitro de complexos cumulus-ovócitos de bovinos
title_fullStr Influência do FSH sobre os aspectos celulares e moleculares na maturação in vitro de complexos cumulus-ovócitos de bovinos
title_full_unstemmed Influência do FSH sobre os aspectos celulares e moleculares na maturação in vitro de complexos cumulus-ovócitos de bovinos
title_sort Influência do FSH sobre os aspectos celulares e moleculares na maturação in vitro de complexos cumulus-ovócitos de bovinos
author CANTANHÊDE, Ludymila Furtado
author_facet CANTANHÊDE, Ludymila Furtado
author_role author
dc.contributor.advisor1.fl_str_mv OLIVEIRA, Marcos Antonio Lemos de
dc.contributor.advisor-co1.fl_str_mv MOURA, Marcelo Tigre
dc.contributor.referee1.fl_str_mv MOURA, Marcelo Tigre
dc.contributor.referee2.fl_str_mv BARTOLOMEU, Cláudio Coutinho
dc.contributor.referee3.fl_str_mv TEIXEIRA, Álvaro Aguiar Coelho
dc.contributor.authorLattes.fl_str_mv http://lattes.cnpq.br/7950707554487640
dc.contributor.author.fl_str_mv CANTANHÊDE, Ludymila Furtado
contributor_str_mv OLIVEIRA, Marcos Antonio Lemos de
MOURA, Marcelo Tigre
MOURA, Marcelo Tigre
BARTOLOMEU, Cláudio Coutinho
TEIXEIRA, Álvaro Aguiar Coelho
dc.subject.por.fl_str_mv Maturação ovocitária
Hormônio folículo estimulante
Bovino
topic Maturação ovocitária
Hormônio folículo estimulante
Bovino
CIENCIAS AGRARIAS::MEDICINA VETERINARIA
dc.subject.cnpq.fl_str_mv CIENCIAS AGRARIAS::MEDICINA VETERINARIA
description Biotechniques applied to bovine reproduction still have limitations and many are linked to a lack of knowledge about an update of macromolecules added to the medium of in vitro oocyte maturation (IVM), embryonic development process and cell difference, as well as genes of inhibitors related to pluripotency in in vitro production in this species. Follicle stimulating hormone (FSH) is present in most IVM protocols because of its importance in large cumulus cells, estradiol synthesis and cytoplasmic and nuclear maturation of the oocyte. However, few studies report an influence of FSH on gene activity during maturation. Considering the identification and modification of the biochemistry of the medium and mechanisms of action of the genes contributed to improvements in the various biotechniques applied to the reproduction to increase the production of animals of high genetic value, this work had the objective of evaluating an influence of FSH in cumulus during maturation of bovine oocytes. Hormonal and biochemical analyzes of the in vitro maturation medium before and MAB, with and FSH (G-FSH, G-FSHIN, G-CTL, G-CTLIN), as well as an analysis of the expression of pluripotency related genes in cumulus of cattle matured with and without FSH (G-FSH and G-CTL) collected in slaughterhouses of the Zona da Mata and Agreste of Pernambuco. A concentration of estradiol was observed by immunoassay and concentration of protein, glycogen, sugar and lipid (biochemical analysis) through a spectrophotometer in the analysis of real-time PCR (RT-qPCR). For statistical analysis, normality was assessed by Shapiro-Wilk test to be submitted to analysis of variance (Test F) and, or Tukey HSD test, level of significance of 5.0%. Increased cumulus cell amplitudes were observed during maturation in the FSH medium when compared to the non-FSH medium. In addition, FSH influences glycogen consumption by cells, decreasing their concentration not medium, due to the need for high energy demand for the process. Influence was observed in the expression of RONIN and CMYC, in which the hormone stimulates RONIN activity and represses CMYC. Thus, FSH influences a maturation of oocytes, besides also acting in the energetic activity of the cumulus-oocyte complex of bovine without cellular and molecular physics.
publishDate 2018
dc.date.accessioned.fl_str_mv 2018-09-04T12:56:30Z
dc.date.issued.fl_str_mv 2018-02-23
dc.type.status.fl_str_mv info:eu-repo/semantics/publishedVersion
dc.type.driver.fl_str_mv info:eu-repo/semantics/doctoralThesis
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dc.identifier.citation.fl_str_mv CANTANHÊDE, Ludymila Furtado. Influência do FSH sobre os aspectos celulares e moleculares na maturação in vitro de complexos cumulus-ovócitos de bovinos. 2018. 114 f. Tese (Programa de Pós-Graduação em Ciência Veterinária) - Universidade Federal Rural de Pernambuco, Recife.
dc.identifier.uri.fl_str_mv http://www.tede2.ufrpe.br:8080/tede2/handle/tede2/7507
identifier_str_mv CANTANHÊDE, Ludymila Furtado. Influência do FSH sobre os aspectos celulares e moleculares na maturação in vitro de complexos cumulus-ovócitos de bovinos. 2018. 114 f. Tese (Programa de Pós-Graduação em Ciência Veterinária) - Universidade Federal Rural de Pernambuco, Recife.
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dc.publisher.country.fl_str_mv Brasil
dc.publisher.department.fl_str_mv Departamento de Medicina Veterinária
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