Rotas de sinalização na divergência folicular e luteólise em bovinos

Detalhes bibliográficos
Autor(a) principal: Rovani, Monique Tomazele
Data de Publicação: 2014
Tipo de documento: Tese
Idioma: por
Título da fonte: Manancial - Repositório Digital da UFSM
Texto Completo: http://repositorio.ufsm.br/handle/1/4110
Resumo: It is well established that locally produced factors exert pivotal roles during dominant follicle selection, oocyte maturation, ovulation and luteolysis. However, the identification of these factors and pathways involved in these processes are not yet established. In the present study, we focused on the in vivo bovine models to study reproductive physiology, which were used to identify receptors and intracellular signaling pathways involved in follicle selection and luteolysis. In the first study, it was reviewed the in vivo models used in our lab, describing and discussing the different bovine models and techniques currently used to study ovarian physiology in this mono-ovulatory specie. In a second study, it was evaluated the expression of estrogen receptors (ESRs) before (day 2 of follicular wave), during (day 3) and after (day 4) follicular deviation in cattle. ESR1 and ESR2 transcripts levels were higher in dominant (F1) than subordinate (F2) follicle after follicular deviation. FSH treatment maintained mRNA levels of both ESR1 and ESR2 in F2 follicles at similar levels observed in F1 follicles. Intrafollicular injection of 100 μM fulvestrant (an antagonist of ESRs) inhibited follicular growth and decreased CYP19A1 mRNA levels. Transcript levels of both ESR1 and ESR2 were not affected by fulvestrant injection. In the third study, our objective was to demonstrate the role of the transcription factor signal transducer and activator of transcription 3 (STAT3) and the nuclear receptor 5A2 (NR5A2) in luteolysis. Luteal and blood samples were collected from separate groups of cows on Day 10 of the estrous cycle 0, 2, 12, 24, and 48 hours after prostaglandin F2 alpha (PGF) treatment. Serum progesterone concentrations decreased (P < 0.05) within 2h and the histological examination of the corpus luteum at 24 and 48h after PGF treatment confirmed functional and morphological luteolysis, respectively. The abundance of STAR mRNA and protein decreased at 12h after PGF treatment. The abundance of NR5A2 mRNA and protein decreased (P < 0.05) at 12 and 24h post-PGF, respectively. Levels of STAT3 mRNA remained constant (P > 0.05) throughout the time-points evaluated. However, the abundance of phosphorylated isoform of STAT3, normalized to total STAT3, increased reaching a peak at 12h and remaining high until 48h after PGF treatment. In conclusion, bovine in vivo models provide a valuable system to study reproductive events under physiological endocrine environment while keeping intact the communication between follicular cells through autocrine and paracrine signaling, without the need to perform ovariectomy or euthanaze the animals. Our results suggest that both ESR1 and ESR2 are regulated during follicular deviation and dominance and in response to FSH treatment in cattle, ESRs are required for normal gene expression and development of the dominant follicle. PGF treatment results in decreased expression of the nuclear receptor NR5A2 and activation of STAT3 by phosphorylation in bovine luteal cells.
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spelling Rotas de sinalização na divergência folicular e luteólise em bovinosSignaling pathways during follicular deviation and luteolysis in cattleBovinosGranulosaEstradiolCorpo lúteoProstaglandina F2αSTAT3NR5A2BovineGranulosaEstradiolCorpus luteumProstaglandin F2αCNPQ::CIENCIAS AGRARIAS::MEDICINA VETERINARIAIt is well established that locally produced factors exert pivotal roles during dominant follicle selection, oocyte maturation, ovulation and luteolysis. However, the identification of these factors and pathways involved in these processes are not yet established. In the present study, we focused on the in vivo bovine models to study reproductive physiology, which were used to identify receptors and intracellular signaling pathways involved in follicle selection and luteolysis. In the first study, it was reviewed the in vivo models used in our lab, describing and discussing the different bovine models and techniques currently used to study ovarian physiology in this mono-ovulatory specie. In a second study, it was evaluated the expression of estrogen receptors (ESRs) before (day 2 of follicular wave), during (day 3) and after (day 4) follicular deviation in cattle. ESR1 and ESR2 transcripts levels were higher in dominant (F1) than subordinate (F2) follicle after follicular deviation. FSH treatment maintained mRNA levels of both ESR1 and ESR2 in F2 follicles at similar levels observed in F1 follicles. Intrafollicular injection of 100 μM fulvestrant (an antagonist of ESRs) inhibited follicular growth and decreased CYP19A1 mRNA levels. Transcript levels of both ESR1 and ESR2 were not affected by fulvestrant injection. In the third study, our objective was to demonstrate the role of the transcription factor signal transducer and activator of transcription 3 (STAT3) and the nuclear receptor 5A2 (NR5A2) in luteolysis. Luteal and blood samples were collected from separate groups of cows on Day 10 of the estrous cycle 0, 2, 12, 24, and 48 hours after prostaglandin F2 alpha (PGF) treatment. Serum progesterone concentrations decreased (P < 0.05) within 2h and the histological examination of the corpus luteum at 24 and 48h after PGF treatment confirmed functional and morphological luteolysis, respectively. The abundance of STAR mRNA and protein decreased at 12h after PGF treatment. The abundance of NR5A2 mRNA and protein decreased (P < 0.05) at 12 and 24h post-PGF, respectively. Levels of STAT3 mRNA remained constant (P > 0.05) throughout the time-points evaluated. However, the abundance of phosphorylated isoform of STAT3, normalized to total STAT3, increased reaching a peak at 12h and remaining high until 48h after PGF treatment. In conclusion, bovine in vivo models provide a valuable system to study reproductive events under physiological endocrine environment while keeping intact the communication between follicular cells through autocrine and paracrine signaling, without the need to perform ovariectomy or euthanaze the animals. Our results suggest that both ESR1 and ESR2 are regulated during follicular deviation and dominance and in response to FSH treatment in cattle, ESRs are required for normal gene expression and development of the dominant follicle. PGF treatment results in decreased expression of the nuclear receptor NR5A2 and activation of STAT3 by phosphorylation in bovine luteal cells.Coordenação de Aperfeiçoamento de Pessoal de Nível SuperiorÉ bem estabelecido que fatores produzidos localmente exercem papel essencial durante a seleção do folículo dominante, maturação oocitária, ovulação e luteólise. No entanto, os fatores e vias envolvidas nestes processos não estão totalmente estabelecidos. No presente estudo, enfatizou-se o uso de modelos bovinos in vivo para o estudo da fisiologia reprodutiva, sendo aqui utilizados para identificar receptores e vias de sinalização intracelular envolvidas na seleção do folículo e luteólise. No primeiro estudo, revisaram-se os modelos in vivo utilizados em nosso laboratório, descreveram-se e discutiram-se os diferentes modelos em bovinos e técnicas atualmente utilizadas para estudar fisiologia ovariana nesta espécie monovulatória. Em um segundo estudo, avaliou-se a expressão de receptores de estradiol (ESRS) antes (dia 2 da onda folicular), durante (dia 3) e após (dia 4) a divergência folicular em bovinos. Os níveis dos transcritos ESR1 e ESR2 foram maiores no folículo dominante (F1) que no subordinado (F2) após a divergência folicular. O tratamento com FSH manteve os níveis de RNAm de ambos ESR1 e ESR2 nos folículos F2 em níveis semelhantes aos observados em folículos F1. A injeção intrafolicular de 100 uM de fulvestrant (um antagonista de ESRs) inibiu o crescimento folicular e causou uma diminuição dos níveis de RNAm de CYP19A1. Os níveis de transcritos, tanto para ESR1 e ESR2, não foram afetados pela injeção de fulvestrant. Num terceiro estudo, o nosso objetivo foi demonstrar o papel do Transdutor de sinais e ativador de transcrição 3 (STAT3) e do receptor nuclear 5A2 (NR5A2) na luteólise. Amostras de corpo lúteo (CL) e sangue foram coletadas dos grupos de vacas 0, 2, 12, 24 e 48 horas após o tratamento com prostaglandina F2 alpha (PGF) no dia 10 do ciclo estral. A concentração de progesterona sérica diminuiu (P < 0.05) em 2 horas e o exame histológico do CL às 24h e 48h após o tratamento com PGF confirmou a ocorrência de luteólise funcional e morfológica, respectivamente. A abundância de RNAm e proteína de STAR diminuiu às 12h após o tratamento com PGF. A abundância de RNAm e proteína de NR5A2 diminuiu (P < 0.05) às 12 e 24 horas pós-PGF, respectivamente. Os níveis de RNAm de STAT3 permaneceram constantes (P> 0.05) ao longo do tempo avaliado. No entanto, a abundância da isoforma fosforilada de STAT3, normalizados para STAT3 total, aumentou, atingindo um pico às 12h e permaneceu elevada até 48h após o tratamento com PGF. Em conclusão, os modelos bovinos in vivo fornecem um sistema valioso para estudar os eventos reprodutivos sob ambiente fisiológico, mantendo intacta a comunicação entre as células foliculares através de sinalização autócrina e parácrina, reduzindo a necessidade de realizar ovariectomia ou realizar a eutanásia dos animais. Nossos resultados sugerem que tanto ESR1 como ESR2 são regulados durante a divergência e dominância folicular em bovinos e em resposta ao tratamento com FSH, e ESRs são necessários para a expressão gênica e para o desenvolvimento do folículo dominante. O tratamento com PGF resulta em diminuição da expressão do receptor nuclear NR5A2 e ativação de STAT3 por fosforilação em células luteais bovinas.Universidade Federal de Santa MariaBRMedicina VeterináriaUFSMPrograma de Pós-Graduação em Medicina VeterináriaGonçalves, Paulo Bayard Diashttp://lattes.cnpq.br/5837260966665885Antoniazzi, Alfredo Quiteshttp://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4162414A9Comim, Fabio Vasconcelloshttp://lattes.cnpq.br/5119233991388822Henkes, Luiz Ernanihttp://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4784025D2Barreta, Marcos Henriquehttp://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4778988E2Rovani, Monique Tomazele2015-11-062015-11-062014-09-12info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/doctoralThesisapplication/pdfapplication/pdfROVANI, Monique Tomazele. SIGNALING PATHWAYS DURING FOLLICULAR DEVIATION AND LUTEOLYSIS IN CATTLE. 2014. 98 f. Tese (Doutorado em Medicina Veterinária) - Universidade Federal de Santa Maria, Santa Maria, 2014.http://repositorio.ufsm.br/handle/1/4110porinfo:eu-repo/semantics/openAccessreponame:Manancial - Repositório Digital da UFSMinstname:Universidade Federal de Santa Maria (UFSM)instacron:UFSM2021-09-23T16:08:45Zoai:repositorio.ufsm.br:1/4110Biblioteca Digital de Teses e Dissertaçõeshttps://repositorio.ufsm.br/ONGhttps://repositorio.ufsm.br/oai/requestatendimento.sib@ufsm.br||tedebc@gmail.comopendoar:2021-09-23T16:08:45Manancial - Repositório Digital da UFSM - Universidade Federal de Santa Maria (UFSM)false
dc.title.none.fl_str_mv Rotas de sinalização na divergência folicular e luteólise em bovinos
Signaling pathways during follicular deviation and luteolysis in cattle
title Rotas de sinalização na divergência folicular e luteólise em bovinos
spellingShingle Rotas de sinalização na divergência folicular e luteólise em bovinos
Rovani, Monique Tomazele
Bovinos
Granulosa
Estradiol
Corpo lúteo
Prostaglandina F2α
STAT3
NR5A2
Bovine
Granulosa
Estradiol
Corpus luteum
Prostaglandin F2α
CNPQ::CIENCIAS AGRARIAS::MEDICINA VETERINARIA
title_short Rotas de sinalização na divergência folicular e luteólise em bovinos
title_full Rotas de sinalização na divergência folicular e luteólise em bovinos
title_fullStr Rotas de sinalização na divergência folicular e luteólise em bovinos
title_full_unstemmed Rotas de sinalização na divergência folicular e luteólise em bovinos
title_sort Rotas de sinalização na divergência folicular e luteólise em bovinos
author Rovani, Monique Tomazele
author_facet Rovani, Monique Tomazele
author_role author
dc.contributor.none.fl_str_mv Gonçalves, Paulo Bayard Dias
http://lattes.cnpq.br/5837260966665885
Antoniazzi, Alfredo Quites
http://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4162414A9
Comim, Fabio Vasconcellos
http://lattes.cnpq.br/5119233991388822
Henkes, Luiz Ernani
http://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4784025D2
Barreta, Marcos Henrique
http://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4778988E2
dc.contributor.author.fl_str_mv Rovani, Monique Tomazele
dc.subject.por.fl_str_mv Bovinos
Granulosa
Estradiol
Corpo lúteo
Prostaglandina F2α
STAT3
NR5A2
Bovine
Granulosa
Estradiol
Corpus luteum
Prostaglandin F2α
CNPQ::CIENCIAS AGRARIAS::MEDICINA VETERINARIA
topic Bovinos
Granulosa
Estradiol
Corpo lúteo
Prostaglandina F2α
STAT3
NR5A2
Bovine
Granulosa
Estradiol
Corpus luteum
Prostaglandin F2α
CNPQ::CIENCIAS AGRARIAS::MEDICINA VETERINARIA
description It is well established that locally produced factors exert pivotal roles during dominant follicle selection, oocyte maturation, ovulation and luteolysis. However, the identification of these factors and pathways involved in these processes are not yet established. In the present study, we focused on the in vivo bovine models to study reproductive physiology, which were used to identify receptors and intracellular signaling pathways involved in follicle selection and luteolysis. In the first study, it was reviewed the in vivo models used in our lab, describing and discussing the different bovine models and techniques currently used to study ovarian physiology in this mono-ovulatory specie. In a second study, it was evaluated the expression of estrogen receptors (ESRs) before (day 2 of follicular wave), during (day 3) and after (day 4) follicular deviation in cattle. ESR1 and ESR2 transcripts levels were higher in dominant (F1) than subordinate (F2) follicle after follicular deviation. FSH treatment maintained mRNA levels of both ESR1 and ESR2 in F2 follicles at similar levels observed in F1 follicles. Intrafollicular injection of 100 μM fulvestrant (an antagonist of ESRs) inhibited follicular growth and decreased CYP19A1 mRNA levels. Transcript levels of both ESR1 and ESR2 were not affected by fulvestrant injection. In the third study, our objective was to demonstrate the role of the transcription factor signal transducer and activator of transcription 3 (STAT3) and the nuclear receptor 5A2 (NR5A2) in luteolysis. Luteal and blood samples were collected from separate groups of cows on Day 10 of the estrous cycle 0, 2, 12, 24, and 48 hours after prostaglandin F2 alpha (PGF) treatment. Serum progesterone concentrations decreased (P < 0.05) within 2h and the histological examination of the corpus luteum at 24 and 48h after PGF treatment confirmed functional and morphological luteolysis, respectively. The abundance of STAR mRNA and protein decreased at 12h after PGF treatment. The abundance of NR5A2 mRNA and protein decreased (P < 0.05) at 12 and 24h post-PGF, respectively. Levels of STAT3 mRNA remained constant (P > 0.05) throughout the time-points evaluated. However, the abundance of phosphorylated isoform of STAT3, normalized to total STAT3, increased reaching a peak at 12h and remaining high until 48h after PGF treatment. In conclusion, bovine in vivo models provide a valuable system to study reproductive events under physiological endocrine environment while keeping intact the communication between follicular cells through autocrine and paracrine signaling, without the need to perform ovariectomy or euthanaze the animals. Our results suggest that both ESR1 and ESR2 are regulated during follicular deviation and dominance and in response to FSH treatment in cattle, ESRs are required for normal gene expression and development of the dominant follicle. PGF treatment results in decreased expression of the nuclear receptor NR5A2 and activation of STAT3 by phosphorylation in bovine luteal cells.
publishDate 2014
dc.date.none.fl_str_mv 2014-09-12
2015-11-06
2015-11-06
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 ROVANI, Monique Tomazele. SIGNALING PATHWAYS DURING FOLLICULAR DEVIATION AND LUTEOLYSIS IN CATTLE. 2014. 98 f. Tese (Doutorado em Medicina Veterinária) - Universidade Federal de Santa Maria, Santa Maria, 2014.
http://repositorio.ufsm.br/handle/1/4110
identifier_str_mv ROVANI, Monique Tomazele. SIGNALING PATHWAYS DURING FOLLICULAR DEVIATION AND LUTEOLYSIS IN CATTLE. 2014. 98 f. Tese (Doutorado em Medicina Veterinária) - Universidade Federal de Santa Maria, Santa Maria, 2014.
url http://repositorio.ufsm.br/handle/1/4110
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
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