Fator de crescimento de fibroblasto 18 (FGF18) modula a expressão de CTGF em CCOs e embriões bovinos produzidos in vitro

Detalhes bibliográficos
Autor(a) principal: Silva, Elisabeth Schmidt da
Data de Publicação: 2021
Tipo de documento: Dissertação
Idioma: por
Título da fonte: Manancial - Repositório Digital da UFSM
dARK ID: ark:/26339/001300000k84n
Texto Completo: http://repositorio.ufsm.br/handle/1/21273
Resumo: In 1995, the Hippo pathway was initially discovered in Drosophila melanogaster, and since then several researchers have been trying to better understand this pathway. The Hippo pathway is involved in cell proliferation processes, both in ovarian follicular cells in the early embryonic development. The effectors of this pathway YAP e TAZ act as transcription coactivators of cell proliferation and survival genes such as CTGF and CYR61. However, the fibroblast growth factor also induces CTGF expression. However, recent studies by the group found that fibroblast growth factor 18 (FGF18) is present in the fallopian tube during early embryonic development. This led us to think that FGF18 would have some role during embryonic development. Therefore, the aim of the following study was to determine whether FGF18 modulates the Hippo pathway through the expression of target genes for cell proliferation (CTGF and CYR61) during oocyte maturation and early embryonic development. To acquire the results of the objective, three experiments were carried out, with in vitro maturation (IVM) of cumulus oocyte complexes (COCs) in the first and second experiments, with the addition of FGF18 during IVM, and the structures were collected at different times. In the first experiment, gradual concentrations of FGF18 (0, 10 and 100ng / mL) were added and there was a control group without the addition of FGF18, and 6, 12 and 24 hours of maturation were collected. In the second experiment, a concentration of 100ng / mL of FGF18 was added during IVM, there was a control group and the structures were collected at 0, 3, 6 and 9 hours of maturation. In the experiment, three COCs were put to maturation and were divided into three groups. A group considered a control group in which COCs were matured, with in vitro fertilization (IVF) and then in vitro culture (IVC) without the addition of FGF18. A group in which, during maturation, FGF18 was added only during IVM and afterwards fertilization and culture were carried out in media without the addition of FGF18. In a third group, COCs were put to mature, fertilized and, when put to culture, FGF18 was added in the medium. The genes evaluated in the three experiments were the cell proliferation genes, CTGF and CYR61. In experiment one, which aimed to evaluate the addition of gradual concentrations during maturation, there was an increase (p <0.05) in CTGF gene expression in the group treated with 100ng / mL at 12 hours. In experiment two, in which the objective was to assess the concentration of 100ng / mL, there was a decrease (p <0.05) in the treated group compared to the control group at three hours. And in experiment three in which the objective was to evaluate the addition of FGF18 during embryonic development, there was a statistical difference (p <0.05) in the group treated with FGF18 during IVC. Therefore, we conclude that FGF18 modulates CTGF expression in critical periods of oocyte nuclear maturation, cumulus expansion and early embryonic development in cattle.
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spelling Fator de crescimento de fibroblasto 18 (FGF18) modula a expressão de CTGF em CCOs e embriões bovinos produzidos in vitroFibroblast growth factor 18 (FGF18) modulates CTGF expression in COCs and bovine embryos produced in vitroVia HippoDesenvolvimento embrionárioFator de crescimento de fibroblasto 18Hippo pathwayEmbryonic developmentFibroblast growth factor 18CNPQ::CIENCIAS AGRARIAS::MEDICINA VETERINARIAIn 1995, the Hippo pathway was initially discovered in Drosophila melanogaster, and since then several researchers have been trying to better understand this pathway. The Hippo pathway is involved in cell proliferation processes, both in ovarian follicular cells in the early embryonic development. The effectors of this pathway YAP e TAZ act as transcription coactivators of cell proliferation and survival genes such as CTGF and CYR61. However, the fibroblast growth factor also induces CTGF expression. However, recent studies by the group found that fibroblast growth factor 18 (FGF18) is present in the fallopian tube during early embryonic development. This led us to think that FGF18 would have some role during embryonic development. Therefore, the aim of the following study was to determine whether FGF18 modulates the Hippo pathway through the expression of target genes for cell proliferation (CTGF and CYR61) during oocyte maturation and early embryonic development. To acquire the results of the objective, three experiments were carried out, with in vitro maturation (IVM) of cumulus oocyte complexes (COCs) in the first and second experiments, with the addition of FGF18 during IVM, and the structures were collected at different times. In the first experiment, gradual concentrations of FGF18 (0, 10 and 100ng / mL) were added and there was a control group without the addition of FGF18, and 6, 12 and 24 hours of maturation were collected. In the second experiment, a concentration of 100ng / mL of FGF18 was added during IVM, there was a control group and the structures were collected at 0, 3, 6 and 9 hours of maturation. In the experiment, three COCs were put to maturation and were divided into three groups. A group considered a control group in which COCs were matured, with in vitro fertilization (IVF) and then in vitro culture (IVC) without the addition of FGF18. A group in which, during maturation, FGF18 was added only during IVM and afterwards fertilization and culture were carried out in media without the addition of FGF18. In a third group, COCs were put to mature, fertilized and, when put to culture, FGF18 was added in the medium. The genes evaluated in the three experiments were the cell proliferation genes, CTGF and CYR61. In experiment one, which aimed to evaluate the addition of gradual concentrations during maturation, there was an increase (p <0.05) in CTGF gene expression in the group treated with 100ng / mL at 12 hours. In experiment two, in which the objective was to assess the concentration of 100ng / mL, there was a decrease (p <0.05) in the treated group compared to the control group at three hours. And in experiment three in which the objective was to evaluate the addition of FGF18 during embryonic development, there was a statistical difference (p <0.05) in the group treated with FGF18 during IVC. Therefore, we conclude that FGF18 modulates CTGF expression in critical periods of oocyte nuclear maturation, cumulus expansion and early embryonic development in cattle.Conselho Nacional de Pesquisa e Desenvolvimento Científico e Tecnológico - CNPqEm 1995 a via Hippo foi inicialmente descoberta na Drosofila melanogaster, e desde então diversos pesquisadores vêm tentando entender melhor essa via. A via Hippo está relacionada a processos de proliferação celular, tanto em células foliculares ovarianas como no desenvolvimento embrionário. Os efetores dessa via, YAP e TAZ atuam como coativadores de transcrição de genes de proliferação e sobrevivência celular como CTGF e CYR61. Porém o fator de crescimento de fibroblasto também induz a expressão de CTGF. E segundo estudos recentes foi descoberto que o fator de crescimento fibroblastico 18 (FGF18) está presente na tuba uterina durante o desenvolvimento embrionário inicial. Isso nos levou a pensar que o FGF18 teria alguma função durante o desenvolvimento embrionário. Portanto, o objetivo deste estudo foi determinar se o FGF18 modula a via Hippo através da expressão de genes alvos de proliferação celular (CTGF e CYR61) durante a maturação oocitária e desenvolvimento embrionário inicial. Para isso três experimentos foram realizados. No primeiro experimento complexos cumulus oocito (CCOs) de bovinos foram maturados in vitro durante 6, 12 e 24 horas na presença de concentrações graduais de FGF18(0, 10 e 100ng/mL). No segundo experimento os CCOs foram maturados 3, 6 e 9 horas na presença de 0 ou 100ng/mL de FGF18. No experimento três CCOs foram colocados para maturação e foram divididos em três grupos. Um grupo considerado grupo controle no qual CCOs foram maturados, realizada a fertilização in vitro (FIV) e depois cultivados in vitro (CIV) sem a adição de FGF18. Um grpo no qual 100nf/mL de FGF18 foram adicionados durante as 24 horas da maturação in vitro e epós foi realizado a fertilização e o cultivo em meios sem adição de FGF18. E um terceiro grupos os CCOs foram cultivados (CIV) por sete dias na presença de 100ng/mL de FGF18. Neste grupo a MIV e FIV foram realizadas na ausência de FGF18. Os genes avaliados nos três experimentos foram os genes de proliferação celular, CTGF e CYR61. O experimento um que teve o objetivo de avaliar a adição de concentrações graduais durante a maturação houve um aumento (p<0,05) da expressão gênica para CTGF no grupo tratado com 100ng/mL as 12horas. No experimento dois em que o objetivo era avaliar a concentração de 100ng/mL houve uma diminuição (p<0,05) do grupo tratado em relação ao grupo controle as três horas. E no experimento três em que o objetivo era avaliar a adição de FGF18 durante o desenvolvimento embrionário, houve diferença estatística (p<0,05) do grupo tratado com FGF18 durante a CIV. Desde forma concluímos que FG18 modula a expressão de CTGF em período críticos da maturação nuclear do oócito, da expansão do cumulus e do desenvolvimento embrionário inicial.Universidade Federal de Santa MariaBrasilMedicina VeterináriaUFSMPrograma de Pós-Graduação em Medicina VeterináriaCentro de Ciências RuraisGonçalves, Paulo Bayard Diashttp://lattes.cnpq.br/5837260966665885Portela Junior, Valério Valdetar MarquesBarreta, Marcos HenriqueFerst, Juliana GermanoSilva, Elisabeth Schmidt da2021-06-30T18:12:38Z2021-06-30T18:12:38Z2021-03-01info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/masterThesisapplication/pdfhttp://repositorio.ufsm.br/handle/1/21273ark:/26339/001300000k84nporAttribution-NonCommercial-NoDerivatives 4.0 Internationalinfo:eu-repo/semantics/openAccessreponame:Manancial - Repositório Digital da UFSMinstname:Universidade Federal de Santa Maria (UFSM)instacron:UFSM2021-07-01T06:02:10Zoai:repositorio.ufsm.br:1/21273Biblioteca Digital de Teses e Dissertaçõeshttps://repositorio.ufsm.br/PUBhttps://repositorio.ufsm.br/oai/requestatendimento.sib@ufsm.br||tedebc@gmail.com||manancial@ufsm.bropendoar:2021-07-01T06:02:10Manancial - Repositório Digital da UFSM - Universidade Federal de Santa Maria (UFSM)false
dc.title.none.fl_str_mv Fator de crescimento de fibroblasto 18 (FGF18) modula a expressão de CTGF em CCOs e embriões bovinos produzidos in vitro
Fibroblast growth factor 18 (FGF18) modulates CTGF expression in COCs and bovine embryos produced in vitro
title Fator de crescimento de fibroblasto 18 (FGF18) modula a expressão de CTGF em CCOs e embriões bovinos produzidos in vitro
spellingShingle Fator de crescimento de fibroblasto 18 (FGF18) modula a expressão de CTGF em CCOs e embriões bovinos produzidos in vitro
Silva, Elisabeth Schmidt da
Via Hippo
Desenvolvimento embrionário
Fator de crescimento de fibroblasto 18
Hippo pathway
Embryonic development
Fibroblast growth factor 18
CNPQ::CIENCIAS AGRARIAS::MEDICINA VETERINARIA
title_short Fator de crescimento de fibroblasto 18 (FGF18) modula a expressão de CTGF em CCOs e embriões bovinos produzidos in vitro
title_full Fator de crescimento de fibroblasto 18 (FGF18) modula a expressão de CTGF em CCOs e embriões bovinos produzidos in vitro
title_fullStr Fator de crescimento de fibroblasto 18 (FGF18) modula a expressão de CTGF em CCOs e embriões bovinos produzidos in vitro
title_full_unstemmed Fator de crescimento de fibroblasto 18 (FGF18) modula a expressão de CTGF em CCOs e embriões bovinos produzidos in vitro
title_sort Fator de crescimento de fibroblasto 18 (FGF18) modula a expressão de CTGF em CCOs e embriões bovinos produzidos in vitro
author Silva, Elisabeth Schmidt da
author_facet Silva, Elisabeth Schmidt da
author_role author
dc.contributor.none.fl_str_mv Gonçalves, Paulo Bayard Dias
http://lattes.cnpq.br/5837260966665885
Portela Junior, Valério Valdetar Marques
Barreta, Marcos Henrique
Ferst, Juliana Germano
dc.contributor.author.fl_str_mv Silva, Elisabeth Schmidt da
dc.subject.por.fl_str_mv Via Hippo
Desenvolvimento embrionário
Fator de crescimento de fibroblasto 18
Hippo pathway
Embryonic development
Fibroblast growth factor 18
CNPQ::CIENCIAS AGRARIAS::MEDICINA VETERINARIA
topic Via Hippo
Desenvolvimento embrionário
Fator de crescimento de fibroblasto 18
Hippo pathway
Embryonic development
Fibroblast growth factor 18
CNPQ::CIENCIAS AGRARIAS::MEDICINA VETERINARIA
description In 1995, the Hippo pathway was initially discovered in Drosophila melanogaster, and since then several researchers have been trying to better understand this pathway. The Hippo pathway is involved in cell proliferation processes, both in ovarian follicular cells in the early embryonic development. The effectors of this pathway YAP e TAZ act as transcription coactivators of cell proliferation and survival genes such as CTGF and CYR61. However, the fibroblast growth factor also induces CTGF expression. However, recent studies by the group found that fibroblast growth factor 18 (FGF18) is present in the fallopian tube during early embryonic development. This led us to think that FGF18 would have some role during embryonic development. Therefore, the aim of the following study was to determine whether FGF18 modulates the Hippo pathway through the expression of target genes for cell proliferation (CTGF and CYR61) during oocyte maturation and early embryonic development. To acquire the results of the objective, three experiments were carried out, with in vitro maturation (IVM) of cumulus oocyte complexes (COCs) in the first and second experiments, with the addition of FGF18 during IVM, and the structures were collected at different times. In the first experiment, gradual concentrations of FGF18 (0, 10 and 100ng / mL) were added and there was a control group without the addition of FGF18, and 6, 12 and 24 hours of maturation were collected. In the second experiment, a concentration of 100ng / mL of FGF18 was added during IVM, there was a control group and the structures were collected at 0, 3, 6 and 9 hours of maturation. In the experiment, three COCs were put to maturation and were divided into three groups. A group considered a control group in which COCs were matured, with in vitro fertilization (IVF) and then in vitro culture (IVC) without the addition of FGF18. A group in which, during maturation, FGF18 was added only during IVM and afterwards fertilization and culture were carried out in media without the addition of FGF18. In a third group, COCs were put to mature, fertilized and, when put to culture, FGF18 was added in the medium. The genes evaluated in the three experiments were the cell proliferation genes, CTGF and CYR61. In experiment one, which aimed to evaluate the addition of gradual concentrations during maturation, there was an increase (p <0.05) in CTGF gene expression in the group treated with 100ng / mL at 12 hours. In experiment two, in which the objective was to assess the concentration of 100ng / mL, there was a decrease (p <0.05) in the treated group compared to the control group at three hours. And in experiment three in which the objective was to evaluate the addition of FGF18 during embryonic development, there was a statistical difference (p <0.05) in the group treated with FGF18 during IVC. Therefore, we conclude that FGF18 modulates CTGF expression in critical periods of oocyte nuclear maturation, cumulus expansion and early embryonic development in cattle.
publishDate 2021
dc.date.none.fl_str_mv 2021-06-30T18:12:38Z
2021-06-30T18:12:38Z
2021-03-01
dc.type.status.fl_str_mv info:eu-repo/semantics/publishedVersion
dc.type.driver.fl_str_mv info:eu-repo/semantics/masterThesis
format masterThesis
status_str publishedVersion
dc.identifier.uri.fl_str_mv http://repositorio.ufsm.br/handle/1/21273
dc.identifier.dark.fl_str_mv ark:/26339/001300000k84n
url http://repositorio.ufsm.br/handle/1/21273
identifier_str_mv ark:/26339/001300000k84n
dc.language.iso.fl_str_mv por
language por
dc.rights.driver.fl_str_mv Attribution-NonCommercial-NoDerivatives 4.0 International
info:eu-repo/semantics/openAccess
rights_invalid_str_mv Attribution-NonCommercial-NoDerivatives 4.0 International
eu_rights_str_mv openAccess
dc.format.none.fl_str_mv application/pdf
dc.publisher.none.fl_str_mv Universidade Federal de Santa Maria
Brasil
Medicina Veterinária
UFSM
Programa de Pós-Graduação em Medicina Veterinária
Centro de Ciências Rurais
publisher.none.fl_str_mv Universidade Federal de Santa Maria
Brasil
Medicina Veterinária
UFSM
Programa de Pós-Graduação em Medicina Veterinária
Centro de Ciências Rurais
dc.source.none.fl_str_mv reponame:Manancial - Repositório Digital da UFSM
instname:Universidade Federal de Santa Maria (UFSM)
instacron:UFSM
instname_str Universidade Federal de Santa Maria (UFSM)
instacron_str UFSM
institution UFSM
reponame_str Manancial - Repositório Digital da UFSM
collection Manancial - Repositório Digital da UFSM
repository.name.fl_str_mv Manancial - Repositório Digital da UFSM - Universidade Federal de Santa Maria (UFSM)
repository.mail.fl_str_mv atendimento.sib@ufsm.br||tedebc@gmail.com||manancial@ufsm.br
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