Fator de crescimento de fibroblasto 18 (FGF18) modula a expressão de CTGF em CCOs e embriões bovinos produzidos in vitro
| Autor(a) principal: | |
|---|---|
| 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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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. |
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2021 |
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2021-06-30T18:12:38Z 2021-06-30T18:12:38Z 2021-03-01 |
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info:eu-repo/semantics/publishedVersion |
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Universidade Federal de Santa Maria Brasil Medicina Veterinária UFSM Programa de Pós-Graduação em Medicina Veterinária Centro de Ciências Rurais |
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Universidade Federal de Santa Maria Brasil Medicina Veterinária UFSM Programa de Pós-Graduação em Medicina Veterinária Centro de Ciências Rurais |
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reponame:Manancial - Repositório Digital da UFSM instname:Universidade Federal de Santa Maria (UFSM) instacron:UFSM |
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