Interação YAP1-TEAD regula a cascata de sinalização do EGF em células do cumulus em bovinos
Autor(a) principal: | |
---|---|
Data de Publicação: | 2020 |
Tipo de documento: | Dissertação |
Idioma: | por |
Título da fonte: | Biblioteca Digital de Teses e Dissertações do UFSM |
Texto Completo: | http://repositorio.ufsm.br/handle/1/19850 |
Resumo: | The mechanism of cumulus cell (CC) expansion and ovulation in vivo is triggered by LH surge in granulosa cells and CC, which culminating in a series of paracrine and autocrine communications between these ovarian cells. It is well established that granulosa-secreted factors act on CC, stimulating important genes involved in cumulus expansion, oocyte maturation and ovulation. Although several signaling pathways involved in this process are already known, there is no complete understanding in this physiological process. Recently, the Hippo signaling pathway and its effectors (YAP1 and TAZ) have been related to different contexts of ovarian physiology, including the ovulatory process, in mice and humans. However, few studies relating this pathway are available in economic relevant species. The aim of this study was to demonstrate the function of YAP1-TEAD interaction in the modulation of key genes for expansion and ovulatory mechanism, as well as, their regulation in bovine cumulus cells. For this, cumulus-oocyte complexes (COCs) were in vitro cultured during 0 or 12h and subjected to immunofluorescence to demonstrate YAP1 and pYAP1 intracellular localization. Our results suggest that YAP1 translocates from the cytoplasm to the nucleus during in vitro maturation (IVM) time. To elucidate YAP1 regulation, we inhibited EGF receptor (EGFR, with AG 1478) and stimulated COCs with EGF or FSH. Thus, EGFR inhibition results in reduction of YAP1 and CTGF (one of the major YAP1-TEAD target genes) expression. Then, COCs were subjected to different concentrations of verteporfin, an YAP1-TEAD inhibitor, and it was obtained dose-dependent reduction of cumulus expansion (HAS2 and PTX3) and ovulation (ADAM17, EREG and PTGS2) related genes. Based on that, it is suggested that YAP1-TEAD interaction is involved in the expression of key genes for cumulus expansion and ovulation in bovine cumulus cells. Also, YAP1 regulation is mediated through EGFR stimulation, by EGF direct action, or FSH indirect action. |
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2020-03-13T22:04:02Z2020-03-13T22:04:02Z2020-02-07http://repositorio.ufsm.br/handle/1/19850The mechanism of cumulus cell (CC) expansion and ovulation in vivo is triggered by LH surge in granulosa cells and CC, which culminating in a series of paracrine and autocrine communications between these ovarian cells. It is well established that granulosa-secreted factors act on CC, stimulating important genes involved in cumulus expansion, oocyte maturation and ovulation. Although several signaling pathways involved in this process are already known, there is no complete understanding in this physiological process. Recently, the Hippo signaling pathway and its effectors (YAP1 and TAZ) have been related to different contexts of ovarian physiology, including the ovulatory process, in mice and humans. However, few studies relating this pathway are available in economic relevant species. The aim of this study was to demonstrate the function of YAP1-TEAD interaction in the modulation of key genes for expansion and ovulatory mechanism, as well as, their regulation in bovine cumulus cells. For this, cumulus-oocyte complexes (COCs) were in vitro cultured during 0 or 12h and subjected to immunofluorescence to demonstrate YAP1 and pYAP1 intracellular localization. Our results suggest that YAP1 translocates from the cytoplasm to the nucleus during in vitro maturation (IVM) time. To elucidate YAP1 regulation, we inhibited EGF receptor (EGFR, with AG 1478) and stimulated COCs with EGF or FSH. Thus, EGFR inhibition results in reduction of YAP1 and CTGF (one of the major YAP1-TEAD target genes) expression. Then, COCs were subjected to different concentrations of verteporfin, an YAP1-TEAD inhibitor, and it was obtained dose-dependent reduction of cumulus expansion (HAS2 and PTX3) and ovulation (ADAM17, EREG and PTGS2) related genes. Based on that, it is suggested that YAP1-TEAD interaction is involved in the expression of key genes for cumulus expansion and ovulation in bovine cumulus cells. Also, YAP1 regulation is mediated through EGFR stimulation, by EGF direct action, or FSH indirect action.O mecanismo de expansão das células do cumulus (CC) e ovulação in vivo é desencadeado pelo pico de LH nas células da granulosa e CC, culminando com uma série de comunicações parácrinas e autócrinas entre estas células ovarianas. Está estabelecido que fatores secretados pela granulosa atuam sobre as CC estimulando a transcrição de genes importantes para a expansão, maturação oocitária e ovulação. Embora já sejam conhecidas diversas vias de sinalização envolvidas com esse processo, ainda não se tem o completo entendimento. Recentemente, a via de sinalização Hippo e seus efetores (YAP1 e TAZ) vêm sendo relacionados com diferentes contextos da fisiologia ovariana, inclusive com o processo ovulatório, em camundongos e humanos. Porém, ainda são escassos os estudos relacionando essa via com espécies de produção. O objetivo deste estudo foi demonstrar a função da ligação YAP1 com fatores de transcrição TEAD na modulação de genes-chave para a expansão e mecanismo ovulatório nas células do cumulus de bovinos e a sua regulação. Para isso, complexos cumulus-oócito (CCO) foram cultivados in vitro durante 0 e 12h e submetidos a imunofluorescência para demonstrar a localização intracelular de YAP1 e pYAP1. Os resultados sugerem que YAP1 transloca do citoplasma para o núcleo das CC durante o processo de maturação in vitro (MIV). Para elucidar a regulação de YAP1, receptores de EGF (EGFR; com AG 1478) foram inibidos e então, os CCOs foram estimulados com EGF ou FSH. Com isso, foi demonstrado que a inibição de EGFR resulta na redução da expressão de YAP1 e CTGF (um dos principais genes-alvo de YAP1-TEAD) nas CC. Então, os CCOs foram tratados com diferentes concentrações de verteporfina, um inibidor da interação YAP1-TEAD e obteve-se a redução de maneira concentração-dependente de genes-chave para a expansão (HAS2 e PTX3) e ovulação (ADAM17, EREG e PTGS2) nas CC. A interação YAP1-TEAD está envolvida na expressão de genes-chave para a expansão e mecanismo ovulatório em células do cumulus de bovinos e a regulação de YAP1 é mediada através da estimulação de EGFR, por ação direta de EGF, ou indireta de FSH.Conselho Nacional de Pesquisa e Desenvolvimento Científico e Tecnológico - CNPqporUniversidade Federal de Santa MariaCentro de Ciências RuraisPrograma de Pós-Graduação em Medicina VeterináriaUFSMBrasilMedicina VeterináriaAttribution-NonCommercial-NoDerivatives 4.0 Internationalhttp://creativecommons.org/licenses/by-nc-nd/4.0/info:eu-repo/semantics/openAccessComplexo cumulus-oócitoExpansão do cumulusOvárioOvulaçãoCumulus-oocyte complexCumulus expansionOvarianOvulationCNPQ::CIENCIAS AGRARIAS::MEDICINA VETERINARIAInteração YAP1-TEAD regula a cascata de sinalização do EGF em células do cumulus em bovinosYAP1-TEAD interaction regulates EGF signaling cascade in cumulus cells in bovineinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/masterThesisGonçalves, Paulo Bayard Diashttp://lattes.cnpq.br/5837260966665885Barreta, Marcos Henriquehttp://lattes.cnpq.br/5885484214886762Silveira, Juliano Coelho dahttp://lattes.cnpq.br/5755389041675506http://lattes.cnpq.br/5295281984181652Koch, Júlia500500000007600fcd33ccc-f08d-428c-b7da-9d3757cf0a96d58aaf45-d288-4316-a726-59c48ec0a5ea0d370fec-324f-4d7c-8cc3-8deefe7a1e1ef11748e8-6031-4d6e-b3fd-8a76d0dad168reponame:Biblioteca Digital de Teses e Dissertações do UFSMinstname:Universidade Federal de Santa Maria (UFSM)instacron:UFSMLICENSElicense.txtlicense.txttext/plain; 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dc.title.por.fl_str_mv |
Interação YAP1-TEAD regula a cascata de sinalização do EGF em células do cumulus em bovinos |
dc.title.alternative.eng.fl_str_mv |
YAP1-TEAD interaction regulates EGF signaling cascade in cumulus cells in bovine |
title |
Interação YAP1-TEAD regula a cascata de sinalização do EGF em células do cumulus em bovinos |
spellingShingle |
Interação YAP1-TEAD regula a cascata de sinalização do EGF em células do cumulus em bovinos Koch, Júlia Complexo cumulus-oócito Expansão do cumulus Ovário Ovulação Cumulus-oocyte complex Cumulus expansion Ovarian Ovulation CNPQ::CIENCIAS AGRARIAS::MEDICINA VETERINARIA |
title_short |
Interação YAP1-TEAD regula a cascata de sinalização do EGF em células do cumulus em bovinos |
title_full |
Interação YAP1-TEAD regula a cascata de sinalização do EGF em células do cumulus em bovinos |
title_fullStr |
Interação YAP1-TEAD regula a cascata de sinalização do EGF em células do cumulus em bovinos |
title_full_unstemmed |
Interação YAP1-TEAD regula a cascata de sinalização do EGF em células do cumulus em bovinos |
title_sort |
Interação YAP1-TEAD regula a cascata de sinalização do EGF em células do cumulus em bovinos |
author |
Koch, Júlia |
author_facet |
Koch, Júlia |
author_role |
author |
dc.contributor.advisor1.fl_str_mv |
Gonçalves, Paulo Bayard Dias |
dc.contributor.advisor1Lattes.fl_str_mv |
http://lattes.cnpq.br/5837260966665885 |
dc.contributor.referee1.fl_str_mv |
Barreta, Marcos Henrique |
dc.contributor.referee1Lattes.fl_str_mv |
http://lattes.cnpq.br/5885484214886762 |
dc.contributor.referee2.fl_str_mv |
Silveira, Juliano Coelho da |
dc.contributor.referee2Lattes.fl_str_mv |
http://lattes.cnpq.br/5755389041675506 |
dc.contributor.authorLattes.fl_str_mv |
http://lattes.cnpq.br/5295281984181652 |
dc.contributor.author.fl_str_mv |
Koch, Júlia |
contributor_str_mv |
Gonçalves, Paulo Bayard Dias Barreta, Marcos Henrique Silveira, Juliano Coelho da |
dc.subject.por.fl_str_mv |
Complexo cumulus-oócito Expansão do cumulus Ovário Ovulação |
topic |
Complexo cumulus-oócito Expansão do cumulus Ovário Ovulação Cumulus-oocyte complex Cumulus expansion Ovarian Ovulation CNPQ::CIENCIAS AGRARIAS::MEDICINA VETERINARIA |
dc.subject.eng.fl_str_mv |
Cumulus-oocyte complex Cumulus expansion Ovarian Ovulation |
dc.subject.cnpq.fl_str_mv |
CNPQ::CIENCIAS AGRARIAS::MEDICINA VETERINARIA |
description |
The mechanism of cumulus cell (CC) expansion and ovulation in vivo is triggered by LH surge in granulosa cells and CC, which culminating in a series of paracrine and autocrine communications between these ovarian cells. It is well established that granulosa-secreted factors act on CC, stimulating important genes involved in cumulus expansion, oocyte maturation and ovulation. Although several signaling pathways involved in this process are already known, there is no complete understanding in this physiological process. Recently, the Hippo signaling pathway and its effectors (YAP1 and TAZ) have been related to different contexts of ovarian physiology, including the ovulatory process, in mice and humans. However, few studies relating this pathway are available in economic relevant species. The aim of this study was to demonstrate the function of YAP1-TEAD interaction in the modulation of key genes for expansion and ovulatory mechanism, as well as, their regulation in bovine cumulus cells. For this, cumulus-oocyte complexes (COCs) were in vitro cultured during 0 or 12h and subjected to immunofluorescence to demonstrate YAP1 and pYAP1 intracellular localization. Our results suggest that YAP1 translocates from the cytoplasm to the nucleus during in vitro maturation (IVM) time. To elucidate YAP1 regulation, we inhibited EGF receptor (EGFR, with AG 1478) and stimulated COCs with EGF or FSH. Thus, EGFR inhibition results in reduction of YAP1 and CTGF (one of the major YAP1-TEAD target genes) expression. Then, COCs were subjected to different concentrations of verteporfin, an YAP1-TEAD inhibitor, and it was obtained dose-dependent reduction of cumulus expansion (HAS2 and PTX3) and ovulation (ADAM17, EREG and PTGS2) related genes. Based on that, it is suggested that YAP1-TEAD interaction is involved in the expression of key genes for cumulus expansion and ovulation in bovine cumulus cells. Also, YAP1 regulation is mediated through EGFR stimulation, by EGF direct action, or FSH indirect action. |
publishDate |
2020 |
dc.date.accessioned.fl_str_mv |
2020-03-13T22:04:02Z |
dc.date.available.fl_str_mv |
2020-03-13T22:04:02Z |
dc.date.issued.fl_str_mv |
2020-02-07 |
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/1/19850 |
url |
http://repositorio.ufsm.br/handle/1/19850 |
dc.language.iso.fl_str_mv |
por |
language |
por |
dc.relation.cnpq.fl_str_mv |
500500000007 |
dc.relation.confidence.fl_str_mv |
600 |
dc.relation.authority.fl_str_mv |
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dc.rights.driver.fl_str_mv |
Attribution-NonCommercial-NoDerivatives 4.0 International http://creativecommons.org/licenses/by-nc-nd/4.0/ info:eu-repo/semantics/openAccess |
rights_invalid_str_mv |
Attribution-NonCommercial-NoDerivatives 4.0 International http://creativecommons.org/licenses/by-nc-nd/4.0/ |
eu_rights_str_mv |
openAccess |
dc.publisher.none.fl_str_mv |
Universidade Federal de Santa Maria Centro de Ciências Rurais |
dc.publisher.program.fl_str_mv |
Programa de Pós-Graduação em Medicina Veterinária |
dc.publisher.initials.fl_str_mv |
UFSM |
dc.publisher.country.fl_str_mv |
Brasil |
dc.publisher.department.fl_str_mv |
Medicina Veterinária |
publisher.none.fl_str_mv |
Universidade Federal de Santa Maria Centro de Ciências Rurais |
dc.source.none.fl_str_mv |
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