Publicação
Epigenetic changes in equine myometrium at different stages of endometrosis
| Resumo: | A reduction in the contractile activity of the myometrium can result in insufficient cleaning of the uterine lumen and the persistence of endometritis, leading to endometrosis. Studies in humans and other species point out to an important role for epigenetic mechanisms in gene expression regulation in the myometrium. However, in mares, there is a lack of information regarding the expression of enzymes responsible for DNA methylation (DNMTs) and demethylation (TETs) and the importance of these mechanisms. The main objective of this work was to determine the mRNA expression of these enzymes and explore the potential role of epigenetic mechanisms in the expression of genes encoding oxytocin (OXTR) and PGF2α (PTGFR) receptors in the myometrium of mares with endometrosis. Myometria were used in the midluteal (n=23) and follicular (n=20) phases of the estrous cycle. The material was grouped according to the corresponding endometrial classification (Kenney and Doig, 1986): category I (without pathological changes), IIA (mild endometrial fibrosis and/or inflammation), IIB (moderate endometrial fibrosis) and III (extensive endometrial fibrosis). Using qPCR method, the expression of DNMTs (DNMT1, DNMT3A, DNMT3B), TETs (TET1, TET2, TET3) and the expression levels of OXTR and PTGFR in the myometrium were determined. Using the bisulfite DNA pyrosequencing method, quantitative analysis (%) of DNA methylation was performed at specific sites (CpG islands) of two regions (exon 1 and 2) of OXTR gene. PTGFR did not have CpG islands and, thus, could not be analyzed. DNMT1 mRNA expression increased in myometrium in category IIA compared to category I in the follicular phase of the estrous cycle (P<0.05). The expression of TET2 mRNA was upregulated in the myometrium in categories I and IIB in the midluteal phase, compared to the follicular phase of the estrous cycle, also showing a positive regulation in category IIB compared to category I, in both phases (P <0.05). TET3 mRNA expression was increased in the myometrium in category IIA compared with category I in the follicular phase and was upregulated in category IIB in the follicular phase when compared with the midluteal phase of the estrous cycle (P<0.05). On the other hand, expression levels of OXTR and PTGFR and the global methylation (%) of OXTR in CpG islands did not show any significant differences between categories. However, an increase in DNA methylation was documented at position 6 of exon 2 of OXTR. The results suggest the potential regulatory role of epigenetic mechanisms in gene expression in the myometrium, related to changes in uterine contraction in mares with endometrosis |
|---|---|
| Autores principais: | Silva, Beatriz de Oliveira Celeiro e |
| Assunto: | Endometrosis Myometrium Epigenetics DNA Methylation Oxytocin Endometrose Miométrio Epigenética Metilação do DNA Oxitocina |
| Ano: | 2024 |
| País: | Portugal |
| Tipo de documento: | dissertação de mestrado |
| Tipo de acesso: | acesso aberto |
| Instituição associada: | Universidade de Lisboa |
| Idioma: | inglês |
| Origem: | Repositório da Universidade de Lisboa |
| _version_ | 1866811285317877760 |
|---|---|
| author | Silva, Beatriz de Oliveira Celeiro e |
| author_facet | Silva, Beatriz de Oliveira Celeiro e |
| author_role | author |
| contributor_name_str_mv | Szóstek-Mioduchowska, Anna Dias, Graça Maria Leitão Ferreira Repositório Científico de Acesso Aberto da ULisboa |
| country_str | PT |
| creators_json_txt | [{\"Person.name\":\"Silva, Beatriz de Oliveira Celeiro e\"}] |
| datacite.contributors.contributor.contributorName.fl_str_mv | Szóstek-Mioduchowska, Anna Dias, Graça Maria Leitão Ferreira Repositório Científico de Acesso Aberto da ULisboa |
| datacite.creators.creator.creatorName.fl_str_mv | Silva, Beatriz de Oliveira Celeiro e |
| datacite.date.Accepted.fl_str_mv | 2024-03-06T00:00:00Z |
| datacite.date.available.fl_str_mv | 2024-04-10T16:02:22Z |
| datacite.date.embargoed.fl_str_mv | 2024-04-10T16:02:22Z |
| datacite.rights.fl_str_mv | http://purl.org/coar/access_right/c_abf2 |
| datacite.subjects.subject.fl_str_mv | Endometrosis Myometrium Epigenetics DNA Methylation Oxytocin Endometrose Miométrio Epigenética Metilação do DNA Oxitocina |
| datacite.titles.title.fl_str_mv | Epigenetic changes in equine myometrium at different stages of endometrosis |
| dc.contributor.none.fl_str_mv | Szóstek-Mioduchowska, Anna Dias, Graça Maria Leitão Ferreira Repositório Científico de Acesso Aberto da ULisboa |
| dc.creator.none.fl_str_mv | Silva, Beatriz de Oliveira Celeiro e |
| dc.date.Accepted.fl_str_mv | 2024-03-06T00:00:00Z |
| dc.date.available.fl_str_mv | 2024-04-10T16:02:22Z |
| dc.date.embargoed.fl_str_mv | 2024-04-10T16:02:22Z |
| dc.format.none.fl_str_mv | application/pdf |
| dc.identifier.none.fl_str_mv | http://hdl.handle.net/10400.5/30717 |
| dc.language.none.fl_str_mv | eng |
| dc.publisher.none.fl_str_mv | Universidade de Lisboa, Faculdade de Medicina Veterinária |
| dc.rights.cclincense.fl_str_mv | http://creativecommons.org/licenses/by/4.0/ |
| dc.rights.none.fl_str_mv | http://purl.org/coar/access_right/c_abf2 |
| dc.subject.none.fl_str_mv | Endometrosis Myometrium Epigenetics DNA Methylation Oxytocin Endometrose Miométrio Epigenética Metilação do DNA Oxitocina |
| dc.title.fl_str_mv | Epigenetic changes in equine myometrium at different stages of endometrosis |
| dc.type.none.fl_str_mv | http://purl.org/coar/resource_type/c_bdcc |
| description | A reduction in the contractile activity of the myometrium can result in insufficient cleaning of the uterine lumen and the persistence of endometritis, leading to endometrosis. Studies in humans and other species point out to an important role for epigenetic mechanisms in gene expression regulation in the myometrium. However, in mares, there is a lack of information regarding the expression of enzymes responsible for DNA methylation (DNMTs) and demethylation (TETs) and the importance of these mechanisms. The main objective of this work was to determine the mRNA expression of these enzymes and explore the potential role of epigenetic mechanisms in the expression of genes encoding oxytocin (OXTR) and PGF2α (PTGFR) receptors in the myometrium of mares with endometrosis. Myometria were used in the midluteal (n=23) and follicular (n=20) phases of the estrous cycle. The material was grouped according to the corresponding endometrial classification (Kenney and Doig, 1986): category I (without pathological changes), IIA (mild endometrial fibrosis and/or inflammation), IIB (moderate endometrial fibrosis) and III (extensive endometrial fibrosis). Using qPCR method, the expression of DNMTs (DNMT1, DNMT3A, DNMT3B), TETs (TET1, TET2, TET3) and the expression levels of OXTR and PTGFR in the myometrium were determined. Using the bisulfite DNA pyrosequencing method, quantitative analysis (%) of DNA methylation was performed at specific sites (CpG islands) of two regions (exon 1 and 2) of OXTR gene. PTGFR did not have CpG islands and, thus, could not be analyzed. DNMT1 mRNA expression increased in myometrium in category IIA compared to category I in the follicular phase of the estrous cycle (P<0.05). The expression of TET2 mRNA was upregulated in the myometrium in categories I and IIB in the midluteal phase, compared to the follicular phase of the estrous cycle, also showing a positive regulation in category IIB compared to category I, in both phases (P <0.05). TET3 mRNA expression was increased in the myometrium in category IIA compared with category I in the follicular phase and was upregulated in category IIB in the follicular phase when compared with the midluteal phase of the estrous cycle (P<0.05). On the other hand, expression levels of OXTR and PTGFR and the global methylation (%) of OXTR in CpG islands did not show any significant differences between categories. However, an increase in DNA methylation was documented at position 6 of exon 2 of OXTR. The results suggest the potential regulatory role of epigenetic mechanisms in gene expression in the myometrium, related to changes in uterine contraction in mares with endometrosis |
| dirty | 0 |
| eu_rights_str_mv | openAccess |
| format | masterThesis |
| fulltext.url.fl_str_mv | https://repositorio.ulisboa.pt/bitstreams/072fd623-c8a3-4a0d-9f78-1cd13d376650/download |
| funding.funder.alternateName_str_mv | FCT |
| funding.funder.identifier_str_mv | http://doi.org/10.13039/501100001871 |
| funding.funder.name_str_mv | Fundação para a Ciência e a Tecnologia |
| funding.name_str_mv | 3599-PPCDT |
| id | ul_fa47bf57cfa4ef1ef136d489686d33f7 |
| identifier.url.fl_str_mv | http://hdl.handle.net/10400.5/30717 |
| instacron_str | ul |
| institution | Universidade de Lisboa |
| instname_str | Universidade de Lisboa |
| language | eng |
| network_acronym_str | ul |
| network_name_str | Repositório da Universidade de Lisboa |
| oai_identifier_str | oai:repositorio.ulisboa.pt:10400.5/30717 |
| organization_str_mv | urn:organizationAcronym:ul |
| person_str_mv | Silva, Beatriz de Oliveira Celeiro e |
| publishDate | 2024 |
| publisher.none.fl_str_mv | Universidade de Lisboa, Faculdade de Medicina Veterinária |
| reponame_str | Repositório da Universidade de Lisboa |
| repository_id_str | urn:repositoryAcronym:ul |
| service_str_mv | urn:repositoryAcronym:ul |
| spelling | engUniversidade de Lisboa, Faculdade de Medicina Veterináriapt_PTA reduction in the contractile activity of the myometrium can result in insufficient cleaning of the uterine lumen and the persistence of endometritis, leading to endometrosis. Studies in humans and other species point out to an important role for epigenetic mechanisms in gene expression regulation in the myometrium. However, in mares, there is a lack of information regarding the expression of enzymes responsible for DNA methylation (DNMTs) and demethylation (TETs) and the importance of these mechanisms. The main objective of this work was to determine the mRNA expression of these enzymes and explore the potential role of epigenetic mechanisms in the expression of genes encoding oxytocin (OXTR) and PGF2α (PTGFR) receptors in the myometrium of mares with endometrosis. Myometria were used in the midluteal (n=23) and follicular (n=20) phases of the estrous cycle. The material was grouped according to the corresponding endometrial classification (Kenney and Doig, 1986): category I (without pathological changes), IIA (mild endometrial fibrosis and/or inflammation), IIB (moderate endometrial fibrosis) and III (extensive endometrial fibrosis). Using qPCR method, the expression of DNMTs (DNMT1, DNMT3A, DNMT3B), TETs (TET1, TET2, TET3) and the expression levels of OXTR and PTGFR in the myometrium were determined. Using the bisulfite DNA pyrosequencing method, quantitative analysis (%) of DNA methylation was performed at specific sites (CpG islands) of two regions (exon 1 and 2) of OXTR gene. PTGFR did not have CpG islands and, thus, could not be analyzed. DNMT1 mRNA expression increased in myometrium in category IIA compared to category I in the follicular phase of the estrous cycle (P<0.05). The expression of TET2 mRNA was upregulated in the myometrium in categories I and IIB in the midluteal phase, compared to the follicular phase of the estrous cycle, also showing a positive regulation in category IIB compared to category I, in both phases (P <0.05). TET3 mRNA expression was increased in the myometrium in category IIA compared with category I in the follicular phase and was upregulated in category IIB in the follicular phase when compared with the midluteal phase of the estrous cycle (P<0.05). On the other hand, expression levels of OXTR and PTGFR and the global methylation (%) of OXTR in CpG islands did not show any significant differences between categories. However, an increase in DNA methylation was documented at position 6 of exon 2 of OXTR. The results suggest the potential regulatory role of epigenetic mechanisms in gene expression in the myometrium, related to changes in uterine contraction in mares with endometrosisapplication/pdfpt_PTEpigenetic changes in equine myometrium at different stages of endometrosisSilva, Beatriz de Oliveira Celeiro eSzóstek-Mioduchowska, AnnaDias, Graça Maria Leitão FerreiraHostingInstitutionOrganizationalRepositório Científico de Acesso Aberto da ULisboae-mailmailto:repositorio@reitoria.ulisboa.ptrepositorio@reitoria.ulisboa.ptURNurn:tid:2038838452024-04-10T16:02:22Z2024-03-062024-03-06T00:00:00ZHandlehttp://hdl.handle.net/10400.5/30717http://purl.org/coar/access_right/c_abf2open accessEndometrosisMyometriumEpigeneticsDNA MethylationOxytocinEndometroseMiométrioEpigenéticaMetilação do DNAOxitocina2283986 bytesFundação para a Ciência e a TecnologiaEpigenetics modifications in the mare endometrium: Implications on fibrosis pathogenesis in endometrosis3599-PPCDTCrossref Funder IDhttp://doi.org/10.13039/501100001871literaturehttp://purl.org/coar/resource_type/c_bdccmaster thesis2024-03-06http://creativecommons.org/licenses/by/4.0/http://purl.org/coar/access_right/c_abf2application/pdffulltexthttps://repositorio.ulisboa.pt/bitstreams/072fd623-c8a3-4a0d-9f78-1cd13d376650/download |
| spellingShingle | Epigenetic changes in equine myometrium at different stages of endometrosis Silva, Beatriz de Oliveira Celeiro e Endometrosis Myometrium Epigenetics DNA Methylation Oxytocin Endometrose Miométrio Epigenética Metilação do DNA Oxitocina |
| status | SINGLETON |
| subject.fl_str_mv | Endometrosis Myometrium Epigenetics DNA Methylation Oxytocin Endometrose Miométrio Epigenética Metilação do DNA Oxitocina |
| title | Epigenetic changes in equine myometrium at different stages of endometrosis |
| title_full | Epigenetic changes in equine myometrium at different stages of endometrosis |
| title_fullStr | Epigenetic changes in equine myometrium at different stages of endometrosis |
| title_full_unstemmed | Epigenetic changes in equine myometrium at different stages of endometrosis |
| title_short | Epigenetic changes in equine myometrium at different stages of endometrosis |
| title_sort | Epigenetic changes in equine myometrium at different stages of endometrosis |
| topic | Endometrosis Myometrium Epigenetics DNA Methylation Oxytocin Endometrose Miométrio Epigenética Metilação do DNA Oxitocina |
| topic_facet | Endometrosis Myometrium Epigenetics DNA Methylation Oxytocin Endometrose Miométrio Epigenética Metilação do DNA Oxitocina |
| url | http://hdl.handle.net/10400.5/30717 |
| visible | 1 |