Caracterização molecular dos componentes do sistema angiotensina-(1-7) durante a divergência folicular e expressão de genes de reparo da fita dupla de DNA em embriões bovinos
Autor(a) principal: | |
---|---|
Data de Publicação: | 2012 |
Tipo de documento: | Tese |
Idioma: | por |
Título da fonte: | Manancial - Repositório Digital da UFSM |
Texto Completo: | http://repositorio.ufsm.br/handle/1/4061 |
Resumo: | The first study characterized the expression of MAS receptor and key enzymes for Ang-(1-7) production, such as, ACE2, NEP and PEP during follicular development. Furthermore, the regulation of local Ang1-7 system was evaluated after the intrafollicular injection of fulvestrant (an estradiolreceptor inhibitor) in the dominant follicle. Cows were ovariectomized when the size between the largest (F1) and the second largest follicle (F2) was not statistically different (Day 2), slightly different (Day 3), or markedly different (Day 4). The mRNA abundance of genes encoding MAS receptor, ACE2, NEP and PEP was evaluated in the follicular cells from F1 and F2. The mRNA expression of MAS receptor was upregulated in the granulosa cells of F2 after the establishment of follicular deviation (Day 4), while PEP mRNA increased during (Day 3) and after (Day 4) the deviation process. However, the mRNA expression of ACE2 was upregulated in the granulosa cells of F1 during and after the deviation process. The mRNA expression of NEP was not regulated in F1 and F2. The MAS receptor was immunolocated in the granulosa and theca cells of F1 and F2 during follicular deviation. Moreover, MAS receptor gene expression increased when the F1 was treated with the estrogen receptor-antagonist in vivo. In conclusion, the expression profile of MAS receptor, ACE2, NEP and PEP in dominant and subordinate follicles indicated that Ang-(1-7) play a role in the regulation of the follicular dominance in cattle. A second study was performed to investigate the expression of genes that control homologous recombination (HR; 53BP1, ATM, RAD50, RAD51, RAD52, BRCA1, BRCA2 and NBS1), and non-homologous end-joining (NHEJ; KU70, KU80 and DNAPK), DNArepair pathways in bovine embryos with high, intermediate or low developmental competence. We also evaluated whether bovine embryos can respond to DNA double-stranded breaks (DSBs) induced by ultraviolet (UV) irradiation by regulating the expression of genes involved in the HR and NHEJ repair pathways. Embryos with high, intermediate or low developmental competence were selected based on the cleavage time after in vitro fertilization and were removed from in vitro culture before (36 h), during (72 h) and after (96 h) the expected period of embryonic genome activation (EGA). All studied genes were expressed before, during and after the EGA period regardless the developmental competence of the embryos. Higher mRNA expression of 53BP1 and RAD52 was found before EGA in embryos with low developmental competence. Expression of 53BP1, RAD51 and KU70 was downregulated at 72 h and upregulated at 168 h post-fertilization in bovine embryos with DSBs induced by UV irradiation. In conclusion, important genes controlling HR and NHEJ repair pathways are expressed in bovine embryos before, during or after EGA. Lower developmental competence seems to be associated with a higher mRNA expression of 53BP1 and RAD52. Bovine embryos can response to UV-induced DSBs after the EGA but HR and NHEJ repair pathways seem to be particularly regulated at the blastocyst stage. |
id |
UFSM_e1c70c584a887020203dc1157bd8487f |
---|---|
oai_identifier_str |
oai:repositorio.ufsm.br:1/4061 |
network_acronym_str |
UFSM |
network_name_str |
Manancial - Repositório Digital da UFSM |
repository_id_str |
|
spelling |
Caracterização molecular dos componentes do sistema angiotensina-(1-7) durante a divergência folicular e expressão de genes de reparo da fita dupla de DNA em embriões bovinosMolecular characterization of the angiotensin-(1-7) system components during follicular deviation and expression of DNA double-stranded repair genes in bovine embryosMASECA2PEPRecombinação homólogaNHEJHomologous recombinationCNPQ::CIENCIAS AGRARIAS::MEDICINA VETERINARIAThe first study characterized the expression of MAS receptor and key enzymes for Ang-(1-7) production, such as, ACE2, NEP and PEP during follicular development. Furthermore, the regulation of local Ang1-7 system was evaluated after the intrafollicular injection of fulvestrant (an estradiolreceptor inhibitor) in the dominant follicle. Cows were ovariectomized when the size between the largest (F1) and the second largest follicle (F2) was not statistically different (Day 2), slightly different (Day 3), or markedly different (Day 4). The mRNA abundance of genes encoding MAS receptor, ACE2, NEP and PEP was evaluated in the follicular cells from F1 and F2. The mRNA expression of MAS receptor was upregulated in the granulosa cells of F2 after the establishment of follicular deviation (Day 4), while PEP mRNA increased during (Day 3) and after (Day 4) the deviation process. However, the mRNA expression of ACE2 was upregulated in the granulosa cells of F1 during and after the deviation process. The mRNA expression of NEP was not regulated in F1 and F2. The MAS receptor was immunolocated in the granulosa and theca cells of F1 and F2 during follicular deviation. Moreover, MAS receptor gene expression increased when the F1 was treated with the estrogen receptor-antagonist in vivo. In conclusion, the expression profile of MAS receptor, ACE2, NEP and PEP in dominant and subordinate follicles indicated that Ang-(1-7) play a role in the regulation of the follicular dominance in cattle. A second study was performed to investigate the expression of genes that control homologous recombination (HR; 53BP1, ATM, RAD50, RAD51, RAD52, BRCA1, BRCA2 and NBS1), and non-homologous end-joining (NHEJ; KU70, KU80 and DNAPK), DNArepair pathways in bovine embryos with high, intermediate or low developmental competence. We also evaluated whether bovine embryos can respond to DNA double-stranded breaks (DSBs) induced by ultraviolet (UV) irradiation by regulating the expression of genes involved in the HR and NHEJ repair pathways. Embryos with high, intermediate or low developmental competence were selected based on the cleavage time after in vitro fertilization and were removed from in vitro culture before (36 h), during (72 h) and after (96 h) the expected period of embryonic genome activation (EGA). All studied genes were expressed before, during and after the EGA period regardless the developmental competence of the embryos. Higher mRNA expression of 53BP1 and RAD52 was found before EGA in embryos with low developmental competence. Expression of 53BP1, RAD51 and KU70 was downregulated at 72 h and upregulated at 168 h post-fertilization in bovine embryos with DSBs induced by UV irradiation. In conclusion, important genes controlling HR and NHEJ repair pathways are expressed in bovine embryos before, during or after EGA. Lower developmental competence seems to be associated with a higher mRNA expression of 53BP1 and RAD52. Bovine embryos can response to UV-induced DSBs after the EGA but HR and NHEJ repair pathways seem to be particularly regulated at the blastocyst stage.Coordenação de Aperfeiçoamento de Pessoal de Nível SuperiorO primeiro estudo caracterizou a expressão do receptor MAS e de enzimas responsáveis pela produção de Ang-(1-7), tais como, enzima conversora de angiotensina 2 (ACE2), endopeptidase neutra (NEP) e prolil endopeptidase (PEP) durante o desenvolvimento folicular. Além disso, a regulação local do sistema Ang-(1-7) foi avaliada após a injeção intrafolicular de fulvestrant (inibidor do receptor de estradiol) no folículo dominante. As vacas foram ovariectomizadas quando o tamanho entre o maior (F1) e o segundo maior folículo (F2) não era estatisticamente diferente (D2), ligeiramente (D3) ou marcadamente diferente (D4). A expressão de RNAm do receptor MAS, ACE2, NEP e PEP foi avaliada nas células foliculares do F1 e F2. O receptor MAS foi mais expresso nas células da granulosa do F2 após o estabelecimento da divergência folicular (D4), enquanto a expressão de PEP aumentou durante (D3) e após (D4) o processo de divergência. Entretanto, a expressão de ACE2 foi maior nas células da granulosa do F1 durante e após a divergência. A expressão de PEP não foi regulada no F1 e F2. O receptor MAS foi imunolocalizado nas células da teca e granulosa do F1 e F2 durante a divergência folicular. A expressão de RNAm do receptor MAS aumentou quando o F1 foi tratado com fulvestrant in vivo. Em conclusão, o perfil de expressão do receptor MAS, ACE2, NEP e PEP nos folículos dominante e subordinado indicam que a Ang-(1-7) apresenta uma função na regulação da dominância folicular em bovinos. Em um segundo estudo investigamos a expressão de genes que controlam o reparo do DNA através das vias de recombinação homóloga (HR; 53BP1, ATM, RAD50, RAD51, RAD52, BRCA1, BRCA2, NBS1) e união terminal não homóloga (NHEJ; KU70, KU80, DNAPK) em embriões bovinos com alta, média ou baixa competência de desenvolvimento. Foi também avaliado se embriões bovinos podem responder a quebra na fita dupla de DNA (DSBs), induzida por irradiação UV, através da regulação de genes envolvidos nas vias de reparo HR e NHEJ. Embriões com alta, média ou baixa competência de desenvolvimento foram selecionados pelo tempo de clivagem após a fertilização in vitro e foram removidos do cultivo antes (36 h), durante (72 h) ou após (96 h) o momento esperado para a ativação do genoma embrionário (AGE). Todos os genes foram expressos antes, durante e após a AGE independentemente da competência de desenvolvimento dos embriões. A expressão de 53BP1 e RAD52 foi maior antes da AGE em embriões com baixa competência de desenvolvimento. A expressão de 53BP1, RAD51 e KU70 foi mais baixa as 72 h e maior as 168 h pós fertilização em embriões com DSBs induzida por irradiação UV. Em conclusão, genes importantes para o controle das vias de reparo HR e NHEJ são expressos em embriões bovinos independentemente do tempo de cultivo ou da competência de desenvolvimento. A menor competência de desenvolvimento embrionário parece estar associada com maior expressão de 53BP1 e RAD52. Os embriões bovinos respondem a DSBs após a AGE mas as vias HR e NHEJ são reguladas principalmente no estágio de blastocisto.Universidade Federal de Santa MariaBRMedicina VeterináriaUFSMPrograma de Pós-Graduação em Medicina VeterináriaGonçalves, Paulo Bayard Diashttp://lattes.cnpq.br/5837260966665885Mezzalira, Alceuhttp://lattes.cnpq.br/2481625293830872Barreto, Katia Padilhahttp://lattes.cnpq.br/9635724660722753Henkes, Luiz Ernanihttp://lattes.cnpq.br/2667616009482476Portela Junior, Valério Valdetar Marqueshttp://lattes.cnpq.br/0564876468635367Barreta, Marcos Henrique2017-06-142017-06-142012-02-24info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/doctoralThesisapplication/pdfapplication/pdfBARRETA, Marcos Henrique. Molecular characterization of the angiotensin-(1-7) system components during follicular deviation and expression of DNA double-stranded repair genes in bovine embryos. 2012. 99 f. Tese (Doutorado em Medicina Veterinária) - Universidade Federal de Santa Maria, Santa Maria, 2012.http://repositorio.ufsm.br/handle/1/4061porinfo:eu-repo/semantics/openAccessreponame:Manancial - Repositório Digital da UFSMinstname:Universidade Federal de Santa Maria (UFSM)instacron:UFSM2023-05-22T12:38:48Zoai:repositorio.ufsm.br:1/4061Biblioteca Digital de Teses e Dissertaçõeshttps://repositorio.ufsm.br/ONGhttps://repositorio.ufsm.br/oai/requestatendimento.sib@ufsm.br||tedebc@gmail.comopendoar:2023-05-22T12:38:48Manancial - Repositório Digital da UFSM - Universidade Federal de Santa Maria (UFSM)false |
dc.title.none.fl_str_mv |
Caracterização molecular dos componentes do sistema angiotensina-(1-7) durante a divergência folicular e expressão de genes de reparo da fita dupla de DNA em embriões bovinos Molecular characterization of the angiotensin-(1-7) system components during follicular deviation and expression of DNA double-stranded repair genes in bovine embryos |
title |
Caracterização molecular dos componentes do sistema angiotensina-(1-7) durante a divergência folicular e expressão de genes de reparo da fita dupla de DNA em embriões bovinos |
spellingShingle |
Caracterização molecular dos componentes do sistema angiotensina-(1-7) durante a divergência folicular e expressão de genes de reparo da fita dupla de DNA em embriões bovinos Barreta, Marcos Henrique MAS ECA2 PEP Recombinação homóloga NHEJ Homologous recombination CNPQ::CIENCIAS AGRARIAS::MEDICINA VETERINARIA |
title_short |
Caracterização molecular dos componentes do sistema angiotensina-(1-7) durante a divergência folicular e expressão de genes de reparo da fita dupla de DNA em embriões bovinos |
title_full |
Caracterização molecular dos componentes do sistema angiotensina-(1-7) durante a divergência folicular e expressão de genes de reparo da fita dupla de DNA em embriões bovinos |
title_fullStr |
Caracterização molecular dos componentes do sistema angiotensina-(1-7) durante a divergência folicular e expressão de genes de reparo da fita dupla de DNA em embriões bovinos |
title_full_unstemmed |
Caracterização molecular dos componentes do sistema angiotensina-(1-7) durante a divergência folicular e expressão de genes de reparo da fita dupla de DNA em embriões bovinos |
title_sort |
Caracterização molecular dos componentes do sistema angiotensina-(1-7) durante a divergência folicular e expressão de genes de reparo da fita dupla de DNA em embriões bovinos |
author |
Barreta, Marcos Henrique |
author_facet |
Barreta, Marcos Henrique |
author_role |
author |
dc.contributor.none.fl_str_mv |
Gonçalves, Paulo Bayard Dias http://lattes.cnpq.br/5837260966665885 Mezzalira, Alceu http://lattes.cnpq.br/2481625293830872 Barreto, Katia Padilha http://lattes.cnpq.br/9635724660722753 Henkes, Luiz Ernani http://lattes.cnpq.br/2667616009482476 Portela Junior, Valério Valdetar Marques http://lattes.cnpq.br/0564876468635367 |
dc.contributor.author.fl_str_mv |
Barreta, Marcos Henrique |
dc.subject.por.fl_str_mv |
MAS ECA2 PEP Recombinação homóloga NHEJ Homologous recombination CNPQ::CIENCIAS AGRARIAS::MEDICINA VETERINARIA |
topic |
MAS ECA2 PEP Recombinação homóloga NHEJ Homologous recombination CNPQ::CIENCIAS AGRARIAS::MEDICINA VETERINARIA |
description |
The first study characterized the expression of MAS receptor and key enzymes for Ang-(1-7) production, such as, ACE2, NEP and PEP during follicular development. Furthermore, the regulation of local Ang1-7 system was evaluated after the intrafollicular injection of fulvestrant (an estradiolreceptor inhibitor) in the dominant follicle. Cows were ovariectomized when the size between the largest (F1) and the second largest follicle (F2) was not statistically different (Day 2), slightly different (Day 3), or markedly different (Day 4). The mRNA abundance of genes encoding MAS receptor, ACE2, NEP and PEP was evaluated in the follicular cells from F1 and F2. The mRNA expression of MAS receptor was upregulated in the granulosa cells of F2 after the establishment of follicular deviation (Day 4), while PEP mRNA increased during (Day 3) and after (Day 4) the deviation process. However, the mRNA expression of ACE2 was upregulated in the granulosa cells of F1 during and after the deviation process. The mRNA expression of NEP was not regulated in F1 and F2. The MAS receptor was immunolocated in the granulosa and theca cells of F1 and F2 during follicular deviation. Moreover, MAS receptor gene expression increased when the F1 was treated with the estrogen receptor-antagonist in vivo. In conclusion, the expression profile of MAS receptor, ACE2, NEP and PEP in dominant and subordinate follicles indicated that Ang-(1-7) play a role in the regulation of the follicular dominance in cattle. A second study was performed to investigate the expression of genes that control homologous recombination (HR; 53BP1, ATM, RAD50, RAD51, RAD52, BRCA1, BRCA2 and NBS1), and non-homologous end-joining (NHEJ; KU70, KU80 and DNAPK), DNArepair pathways in bovine embryos with high, intermediate or low developmental competence. We also evaluated whether bovine embryos can respond to DNA double-stranded breaks (DSBs) induced by ultraviolet (UV) irradiation by regulating the expression of genes involved in the HR and NHEJ repair pathways. Embryos with high, intermediate or low developmental competence were selected based on the cleavage time after in vitro fertilization and were removed from in vitro culture before (36 h), during (72 h) and after (96 h) the expected period of embryonic genome activation (EGA). All studied genes were expressed before, during and after the EGA period regardless the developmental competence of the embryos. Higher mRNA expression of 53BP1 and RAD52 was found before EGA in embryos with low developmental competence. Expression of 53BP1, RAD51 and KU70 was downregulated at 72 h and upregulated at 168 h post-fertilization in bovine embryos with DSBs induced by UV irradiation. In conclusion, important genes controlling HR and NHEJ repair pathways are expressed in bovine embryos before, during or after EGA. Lower developmental competence seems to be associated with a higher mRNA expression of 53BP1 and RAD52. Bovine embryos can response to UV-induced DSBs after the EGA but HR and NHEJ repair pathways seem to be particularly regulated at the blastocyst stage. |
publishDate |
2012 |
dc.date.none.fl_str_mv |
2012-02-24 2017-06-14 2017-06-14 |
dc.type.status.fl_str_mv |
info:eu-repo/semantics/publishedVersion |
dc.type.driver.fl_str_mv |
info:eu-repo/semantics/doctoralThesis |
format |
doctoralThesis |
status_str |
publishedVersion |
dc.identifier.uri.fl_str_mv |
BARRETA, Marcos Henrique. Molecular characterization of the angiotensin-(1-7) system components during follicular deviation and expression of DNA double-stranded repair genes in bovine embryos. 2012. 99 f. Tese (Doutorado em Medicina Veterinária) - Universidade Federal de Santa Maria, Santa Maria, 2012. http://repositorio.ufsm.br/handle/1/4061 |
identifier_str_mv |
BARRETA, Marcos Henrique. Molecular characterization of the angiotensin-(1-7) system components during follicular deviation and expression of DNA double-stranded repair genes in bovine embryos. 2012. 99 f. Tese (Doutorado em Medicina Veterinária) - Universidade Federal de Santa Maria, Santa Maria, 2012. |
url |
http://repositorio.ufsm.br/handle/1/4061 |
dc.language.iso.fl_str_mv |
por |
language |
por |
dc.rights.driver.fl_str_mv |
info:eu-repo/semantics/openAccess |
eu_rights_str_mv |
openAccess |
dc.format.none.fl_str_mv |
application/pdf application/pdf |
dc.publisher.none.fl_str_mv |
Universidade Federal de Santa Maria BR Medicina Veterinária UFSM Programa de Pós-Graduação em Medicina Veterinária |
publisher.none.fl_str_mv |
Universidade Federal de Santa Maria BR Medicina Veterinária UFSM Programa de Pós-Graduação em Medicina Veterinária |
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 |
_version_ |
1805922121093742592 |