Extracellular vesicles secreted by bovine embryos produced in vivo and in vitro: miRNAs content and molecular effects in endometrial and luteal tissues

Detalhes bibliográficos
Autor(a) principal: Bridi, Alessandra
Data de Publicação: 2021
Tipo de documento: Tese
Idioma: eng
Título da fonte: Biblioteca Digital de Teses e Dissertações da USP
Texto Completo: https://www.teses.usp.br/teses/disponiveis/74/74135/tde-27102021-164258/
Resumo: Metabolic profiles, gene expression patterns, embryonic development, and the ability to establish and sustain early pregnancies are differences founded when in vivo- and in vitro- produced bovine embryos are compared. To the establishment of a successful pregnancy the perfect embryo-maternal communication is needed. Small extracellular vesicles (sEVs) are part of this crosstalk and carry bioactive molecules such as miRNAs that can act on target bovine cells within the reproductive system. Thereby, our hypothesis is that in vivo and in vitro bovine embryos secrete sEVs containing different miRNAs that can differentially modulate endometrial gene expression pattern and miRNAs profile in the corpus luteum (CL). To address this hypothesis, firstly, we investigated the miRNA content of in vivo and in vitro hatched blastocysts and in sEVs secreted by them. Day 9 in vivo or in vitro hatched blastocysts have distinct miRNA profiles. Small EVs secreted by them from day 7 up to day 9 of development contain different miRNAs. These miRNAs differently expressed are predicted to regulate pathways involved with early embryonic development and endometrial receptivity. Thereby, early embryo-maternal interactions can be modified and consequently can affect pregnancy success. Secondly, we investigated changes in the endometrial global transcriptome after the co-culture of endometrial explants with a day 7 in vivo or in vitro blastocysts. Differently expressed genes were identified in the endometrium and are associated with the embryo\'s presence or origin. Moreover, sEVs present in the conditioned media (C.M.) by these embryonic and endometrial cells contain different miRNAs, which are predicted to modulate the oxytocin signaling pathway. Finally, to understand if the communication among embryo, endometrium, and corpus luteum can be mediated by sEVs, luteal explants were treated with sEVs from C.M. by endometrial explants alone or cultured with a day 7 in vivo or in vitro blastocysts. MicroRNAs profile was modified in luteal explants treated with these sEVs, and we can observe an effect of the embryo presence and origin in these alterations. Together these results suggest that sEVs mediate early embryo-endometrium-corpus luteum communication, contributing to maintaining luteal viability and functionality. However, further experiments are needed to evaluate specific biological effects of these miRNAs carried by sEVs in the endometrium and in the corpus luteum. Moreover, the embryo origin (in vivo and in vitro) modify the interactions among embryo-endometrium-corpus luteum, suggesting strongly that these embryos have distinct needed to develop.
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spelling Extracellular vesicles secreted by bovine embryos produced in vivo and in vitro: miRNAs content and molecular effects in endometrial and luteal tissuesVesículas extracelulares secretadas por embriões bovinos produzidos in vivo e in vitro: conteúdo de miRNAs e os efeitos moleculares sobre o endométrio e o corpo lúteoBlastocistoBlastocystCommunicationComunicaçãoCorpo lúteoCorpus luteumEndométrioEndometriumExtracellular vesiclesMicroRNAsMicroRNAsVesículas extracelularesMetabolic profiles, gene expression patterns, embryonic development, and the ability to establish and sustain early pregnancies are differences founded when in vivo- and in vitro- produced bovine embryos are compared. To the establishment of a successful pregnancy the perfect embryo-maternal communication is needed. Small extracellular vesicles (sEVs) are part of this crosstalk and carry bioactive molecules such as miRNAs that can act on target bovine cells within the reproductive system. Thereby, our hypothesis is that in vivo and in vitro bovine embryos secrete sEVs containing different miRNAs that can differentially modulate endometrial gene expression pattern and miRNAs profile in the corpus luteum (CL). To address this hypothesis, firstly, we investigated the miRNA content of in vivo and in vitro hatched blastocysts and in sEVs secreted by them. Day 9 in vivo or in vitro hatched blastocysts have distinct miRNA profiles. Small EVs secreted by them from day 7 up to day 9 of development contain different miRNAs. These miRNAs differently expressed are predicted to regulate pathways involved with early embryonic development and endometrial receptivity. Thereby, early embryo-maternal interactions can be modified and consequently can affect pregnancy success. Secondly, we investigated changes in the endometrial global transcriptome after the co-culture of endometrial explants with a day 7 in vivo or in vitro blastocysts. Differently expressed genes were identified in the endometrium and are associated with the embryo\'s presence or origin. Moreover, sEVs present in the conditioned media (C.M.) by these embryonic and endometrial cells contain different miRNAs, which are predicted to modulate the oxytocin signaling pathway. Finally, to understand if the communication among embryo, endometrium, and corpus luteum can be mediated by sEVs, luteal explants were treated with sEVs from C.M. by endometrial explants alone or cultured with a day 7 in vivo or in vitro blastocysts. MicroRNAs profile was modified in luteal explants treated with these sEVs, and we can observe an effect of the embryo presence and origin in these alterations. Together these results suggest that sEVs mediate early embryo-endometrium-corpus luteum communication, contributing to maintaining luteal viability and functionality. However, further experiments are needed to evaluate specific biological effects of these miRNAs carried by sEVs in the endometrium and in the corpus luteum. Moreover, the embryo origin (in vivo and in vitro) modify the interactions among embryo-endometrium-corpus luteum, suggesting strongly that these embryos have distinct needed to develop.Perfis metabólicos, padrões de expressão gênica, desenvolvimento embrionário, capacidade de estabelecer e manter gestações precoces são diferenças encontradas quando embriões bovinos produzidos in vivo e in vitro são comparados. Para que a gestação tenha sucesso é necessária uma perfeita comunicação entre o embrião e o organismo materno. Vesículas extracelulares pequenas (sEVs) fazem parte dessa comunicação e carregam moléculas bioativas, como miRNAs, que podem atuar em células-alvo dentro do sistema reprodutivo bovino. Com isso, nossa hipótese é que embriões bovinos produzidos in vivo e in vitro secretam sEVs contendo diferentes miRNAs que podem modular diferencialmente o padrão de expressão gênica endometrial e o perfil de miRNAs do corpo lúteo. Para isso, primeiramente, investigamos o conteúdo de miRNA em blastocistos eclodidos produzidos in vivo ou in vitro e, em sEVs secretadas por eles. Blastocistos eclodidos produzidos in vivo ou in vitro, no dia 9 de desenvolvimento, possuem diferentes perfis de miRNA. EVs pequenas secretadas por eles, do dia 7 até o dia 9 de desenvolvimento, contém miRNAs diferentes. Esses miRNAs diferentemente expressos são preditos por regularem vias envolvidas no desenvolvimento embrionário inicial e na receptividade endometrial. Por meio disso, as primeiras interações entre o embrião e o organismo materno podem ser modificadas, e isso pode afetar o sucesso da gestação. Em segundo lugar, investigamos as alterações no transcriptoma global do endométrio após o co-cultivo de explantes endometriais com um blastocisto produzido in vivo ou in vitro no dia 7 do desenvolvimento. Genes diferentemente expressos foram identificados no endométrio e estão associados com a presença e origem do embrião. Além disso, sEVs presentes nos meios condicionados (C.M.) por essas células embrionárias e endometriais contém diferentes miRNAs, que são preditos por modularem a via de sinalização da oxitocina. Finalmente, para compreender melhor se a comunicação entre o embrião, o endométrio e o corpo lúteo pode ser intermediada por sEVs, explantes luteais foram tratados com sEVs presentes nos C.M. por explantes endometriais cultivados sozinhos ou com um blastocisto bovino produzido in vivo ou in vitro no dia 7 do desenvolvimento. O perfil de miRNAs foi modificado nos explantes luteais tratados com essas sEVs e, nós podemos observar um efeito da presença e origem dos embriões nessas alterações. Juntos, esses resultados sugerem que sEVs intermedeiam a comunicação inicial entre o embrião, endométrio e o corpo lúteo, contribuindo para manter a viabilidade e funcionalidade luteal. Entretanto, futuros experimentos precisam ser realizados para avaliar efeitos biológicos específicos desses miRNAs carreados por essas sEVs no endométrio e no corpo lúteo. Além disso, a origem do embrião (in vivo ou in vitro) modifica as interações entre ele, o endométrio e o corpo lúteo, sugerindo fortemente que esses embriões tenham necessidades distintas para se desenvolverem.Biblioteca Digitais de Teses e Dissertações da USPPerecin, FelipeSilveira, Juliano Coelho daBridi, Alessandra2021-09-13info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/doctoralThesisapplication/pdfhttps://www.teses.usp.br/teses/disponiveis/74/74135/tde-27102021-164258/reponame:Biblioteca Digital de Teses e Dissertações da USPinstname:Universidade de São Paulo (USP)instacron:USPLiberar o conteúdo para acesso público.info:eu-repo/semantics/openAccesseng2021-10-29T13:04:02Zoai:teses.usp.br:tde-27102021-164258Biblioteca Digital de Teses e Dissertaçõeshttp://www.teses.usp.br/PUBhttp://www.teses.usp.br/cgi-bin/mtd2br.plvirginia@if.usp.br|| atendimento@aguia.usp.br||virginia@if.usp.bropendoar:27212021-10-29T13:04:02Biblioteca Digital de Teses e Dissertações da USP - Universidade de São Paulo (USP)false
dc.title.none.fl_str_mv Extracellular vesicles secreted by bovine embryos produced in vivo and in vitro: miRNAs content and molecular effects in endometrial and luteal tissues
Vesículas extracelulares secretadas por embriões bovinos produzidos in vivo e in vitro: conteúdo de miRNAs e os efeitos moleculares sobre o endométrio e o corpo lúteo
title Extracellular vesicles secreted by bovine embryos produced in vivo and in vitro: miRNAs content and molecular effects in endometrial and luteal tissues
spellingShingle Extracellular vesicles secreted by bovine embryos produced in vivo and in vitro: miRNAs content and molecular effects in endometrial and luteal tissues
Bridi, Alessandra
Blastocisto
Blastocyst
Communication
Comunicação
Corpo lúteo
Corpus luteum
Endométrio
Endometrium
Extracellular vesicles
MicroRNAs
MicroRNAs
Vesículas extracelulares
title_short Extracellular vesicles secreted by bovine embryos produced in vivo and in vitro: miRNAs content and molecular effects in endometrial and luteal tissues
title_full Extracellular vesicles secreted by bovine embryos produced in vivo and in vitro: miRNAs content and molecular effects in endometrial and luteal tissues
title_fullStr Extracellular vesicles secreted by bovine embryos produced in vivo and in vitro: miRNAs content and molecular effects in endometrial and luteal tissues
title_full_unstemmed Extracellular vesicles secreted by bovine embryos produced in vivo and in vitro: miRNAs content and molecular effects in endometrial and luteal tissues
title_sort Extracellular vesicles secreted by bovine embryos produced in vivo and in vitro: miRNAs content and molecular effects in endometrial and luteal tissues
author Bridi, Alessandra
author_facet Bridi, Alessandra
author_role author
dc.contributor.none.fl_str_mv Perecin, Felipe
Silveira, Juliano Coelho da
dc.contributor.author.fl_str_mv Bridi, Alessandra
dc.subject.por.fl_str_mv Blastocisto
Blastocyst
Communication
Comunicação
Corpo lúteo
Corpus luteum
Endométrio
Endometrium
Extracellular vesicles
MicroRNAs
MicroRNAs
Vesículas extracelulares
topic Blastocisto
Blastocyst
Communication
Comunicação
Corpo lúteo
Corpus luteum
Endométrio
Endometrium
Extracellular vesicles
MicroRNAs
MicroRNAs
Vesículas extracelulares
description Metabolic profiles, gene expression patterns, embryonic development, and the ability to establish and sustain early pregnancies are differences founded when in vivo- and in vitro- produced bovine embryos are compared. To the establishment of a successful pregnancy the perfect embryo-maternal communication is needed. Small extracellular vesicles (sEVs) are part of this crosstalk and carry bioactive molecules such as miRNAs that can act on target bovine cells within the reproductive system. Thereby, our hypothesis is that in vivo and in vitro bovine embryos secrete sEVs containing different miRNAs that can differentially modulate endometrial gene expression pattern and miRNAs profile in the corpus luteum (CL). To address this hypothesis, firstly, we investigated the miRNA content of in vivo and in vitro hatched blastocysts and in sEVs secreted by them. Day 9 in vivo or in vitro hatched blastocysts have distinct miRNA profiles. Small EVs secreted by them from day 7 up to day 9 of development contain different miRNAs. These miRNAs differently expressed are predicted to regulate pathways involved with early embryonic development and endometrial receptivity. Thereby, early embryo-maternal interactions can be modified and consequently can affect pregnancy success. Secondly, we investigated changes in the endometrial global transcriptome after the co-culture of endometrial explants with a day 7 in vivo or in vitro blastocysts. Differently expressed genes were identified in the endometrium and are associated with the embryo\'s presence or origin. Moreover, sEVs present in the conditioned media (C.M.) by these embryonic and endometrial cells contain different miRNAs, which are predicted to modulate the oxytocin signaling pathway. Finally, to understand if the communication among embryo, endometrium, and corpus luteum can be mediated by sEVs, luteal explants were treated with sEVs from C.M. by endometrial explants alone or cultured with a day 7 in vivo or in vitro blastocysts. MicroRNAs profile was modified in luteal explants treated with these sEVs, and we can observe an effect of the embryo presence and origin in these alterations. Together these results suggest that sEVs mediate early embryo-endometrium-corpus luteum communication, contributing to maintaining luteal viability and functionality. However, further experiments are needed to evaluate specific biological effects of these miRNAs carried by sEVs in the endometrium and in the corpus luteum. Moreover, the embryo origin (in vivo and in vitro) modify the interactions among embryo-endometrium-corpus luteum, suggesting strongly that these embryos have distinct needed to develop.
publishDate 2021
dc.date.none.fl_str_mv 2021-09-13
dc.type.status.fl_str_mv info:eu-repo/semantics/publishedVersion
dc.type.driver.fl_str_mv info:eu-repo/semantics/doctoralThesis
format doctoralThesis
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dc.identifier.uri.fl_str_mv https://www.teses.usp.br/teses/disponiveis/74/74135/tde-27102021-164258/
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dc.language.iso.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv
dc.rights.driver.fl_str_mv Liberar o conteúdo para acesso público.
info:eu-repo/semantics/openAccess
rights_invalid_str_mv Liberar o conteúdo para acesso público.
eu_rights_str_mv openAccess
dc.format.none.fl_str_mv application/pdf
dc.coverage.none.fl_str_mv
dc.publisher.none.fl_str_mv Biblioteca Digitais de Teses e Dissertações da USP
publisher.none.fl_str_mv Biblioteca Digitais de Teses e Dissertações da USP
dc.source.none.fl_str_mv
reponame:Biblioteca Digital de Teses e Dissertações da USP
instname:Universidade de São Paulo (USP)
instacron:USP
instname_str Universidade de São Paulo (USP)
instacron_str USP
institution USP
reponame_str Biblioteca Digital de Teses e Dissertações da USP
collection Biblioteca Digital de Teses e Dissertações da USP
repository.name.fl_str_mv Biblioteca Digital de Teses e Dissertações da USP - Universidade de São Paulo (USP)
repository.mail.fl_str_mv virginia@if.usp.br|| atendimento@aguia.usp.br||virginia@if.usp.br
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