Caracterização do sistema calicreína-cinina durante o processo ovulatório de bovinos

Detalhes bibliográficos
Autor(a) principal: Ilha, Gustavo Freitas
Data de Publicação: 2011
Tipo de documento: Dissertação
Idioma: por
Título da fonte: Manancial - Repositório Digital da UFSM
Texto Completo: http://repositorio.ufsm.br/handle/1/10101
Resumo: The kallikrein-kinin system (KKS) has been described as an important mediator of physiologic processes. Kallikreins use kininogen (KNG) as substrate to generate bradykinin, the principal active peptide of the KKS which acts through two types of receptors, the B1R and B2R. The objective of this study was to characterize some components of KKS in different compartments of ovary during the ovulation process in bovine. mRNA expression pattern of KNG, B1R and B2R was assessed in theca and granulosa cells and bradykinin concentration and kallikrein-like activity in follicular fluid of bovine peri-ovulatory follicles. In order to obtain a peri-ovulatory follicle (≥ 12mm), twenty-seven cows were submitted to estrus synchronization protocol and ovariectomized by colpotomy at 0, 3, 6, 12 or 24 hours after a GnRH-analog injection (gonadorelin; 100 μg, IM). Follicular fluid was aspirated for enzymatic assays and granulosa and theca cells were harvested for mRNA analysis. The mRNA expressions in follicular cells were evaluated by real-time RT-PCR and data represented as relative to housekeeping gene cyclophilin. Bradykinin concentration and Kallikrein-like activity was measured in follicular fluid by enzymatic immunoassay and selective substrate cleavage, respectively, and the absorbance measured using a plate reader. KNG mRNA expression was similar for both follicular cell types (P>0.05), while B2R expression in theca cells and B1R expression in theca and granulosa cells showed different profiles during peri-ovulatory period (P<0.05). Bradykinin concentration and kallikrein-like activity in follicular fluid were different (P<0.05) according the time during ovulation process. The results provide an important characterization of the presence and possible regulation of KKS during ovulation in bovine.
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spelling 2017-06-072017-06-072011-02-25ILHA, Gustavo Freitas. Characterization of kallikrein-kinin system during the ovulation process in bovine. 2011. 40 f. Dissertação (Mestrado em Medicina Veterinária) - Universidade Federal de Santa Maria, Santa Maria, 2011.http://repositorio.ufsm.br/handle/1/10101The kallikrein-kinin system (KKS) has been described as an important mediator of physiologic processes. Kallikreins use kininogen (KNG) as substrate to generate bradykinin, the principal active peptide of the KKS which acts through two types of receptors, the B1R and B2R. The objective of this study was to characterize some components of KKS in different compartments of ovary during the ovulation process in bovine. mRNA expression pattern of KNG, B1R and B2R was assessed in theca and granulosa cells and bradykinin concentration and kallikrein-like activity in follicular fluid of bovine peri-ovulatory follicles. In order to obtain a peri-ovulatory follicle (≥ 12mm), twenty-seven cows were submitted to estrus synchronization protocol and ovariectomized by colpotomy at 0, 3, 6, 12 or 24 hours after a GnRH-analog injection (gonadorelin; 100 μg, IM). Follicular fluid was aspirated for enzymatic assays and granulosa and theca cells were harvested for mRNA analysis. The mRNA expressions in follicular cells were evaluated by real-time RT-PCR and data represented as relative to housekeeping gene cyclophilin. Bradykinin concentration and Kallikrein-like activity was measured in follicular fluid by enzymatic immunoassay and selective substrate cleavage, respectively, and the absorbance measured using a plate reader. KNG mRNA expression was similar for both follicular cell types (P>0.05), while B2R expression in theca cells and B1R expression in theca and granulosa cells showed different profiles during peri-ovulatory period (P<0.05). Bradykinin concentration and kallikrein-like activity in follicular fluid were different (P<0.05) according the time during ovulation process. The results provide an important characterization of the presence and possible regulation of KKS during ovulation in bovine.O sistema calicreína-cinina (KKS) tem sido descrito como um importante mediador de processos fisiológicos. Calicreínas utilizam o cininogênio (KNG) como substrato para formar a bradicinina, que é o principal peptídeo ativo do KKS o qual atua através de dois tipos de receptores, o B1R e B2R. O objetivo deste estudo foi caracterizar os principais componentes do KKS em diferentes compartimentos ovarianos durante o processo ovulatório de bovinos. A expressão de RNAm de KNG, B1R e B2R foi mensurada em células da teca e granulosa, e concentração de bradicinina e atividade de calicreína no fluido folicular de folículos peri-ovulatórios bovinos. Para obter um folículo peri-ovulatório (≥ 12mm), vinte e sete vacas foram submetidas a um protocolo de sincronização de cios e ovariectomizadas por colpotomia 0, 3, 6, 12 ou 24 horas após uma injeção de um análogo ao GnRH (gonadorelina; 100 μg, IM). O fluido folicular foi aspirado para os ensaios enzimáticos e as células da teca e granulosa dissecadas para análise do RNAm. A expressão do RNAm em células foliculares foi avaliada por PCR em tempo real e os dados representados em relação ao gene constitutivo ciclofilina. A concentração de bradicinina e atividade de calicreína foram mensuradas no fluido follicular por imunoensaio enzimático e clivagem de substrato seletivo, respectivamente, e sua absorbância mensurada por leitor de placas. A expressão de RNAm para o KNG não variou em ambos os tipos celulares nos diferentes tempos (P>0,05), enquanto que para B2R a expressão em células da teca e expressão para B1R nas células da teca e granulosa apresentaram diferentes padrões durante o período peri-ovulatório (P<0,05). A concentração de bradicinina e a atividade de calicreína no fluido follicular foram diferentes (P<0,05) de acordo com o tempo durante o processo ovulatório. Estes resultados demonstram que o KKS está presente e há indicativos de sua regulação durante a ovulação em bovinosConselho Nacional de Desenvolvimento Científico e Tecnológicoapplication/pdfporUniversidade Federal de Santa MariaPrograma de Pós-Graduação em Medicina VeterináriaUFSMBRMedicina VeterináriaOvulaçãoSistema calicreína-cininaBradicininaOvárioBovinoOvulationKallikrein-kinin systemBradykininOvaryBovineCNPQ::CIENCIAS AGRARIAS::MEDICINA VETERINARIACaracterização do sistema calicreína-cinina durante o processo ovulatório de bovinosCharacterization of kallikrein-kinin system during the ovulation process in bovineinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/masterThesisOliveira, João Francisco Coelho dehttp://lattes.cnpq.br/1168979725730267Mezzalira, Alceuhttp://lattes.cnpq.br/2481625293830872Ferreira, Julianohttp://lattes.cnpq.br/2694197910478313http://lattes.cnpq.br/1907756288556494Ilha, Gustavo Freitas500500000007400500500500500de4bcf39-3a86-4cfa-a9b0-e1ea1ceb741ccf6fc24b-4076-47fd-ae63-caa7ab6f5fa6f6687207-7d57-473d-9743-de391de747aab410c4c0-c1c6-41df-aed8-741731a2b8a2info:eu-repo/semantics/openAccessreponame:Manancial - Repositório Digital da UFSMinstname:Universidade Federal de Santa Maria (UFSM)instacron:UFSMORIGINALILHA, GUSTAVO FREITAS.pdfapplication/pdf229857http://repositorio.ufsm.br/bitstream/1/10101/1/ILHA%2c%20GUSTAVO%20FREITAS.pdf5fb2574a028ee182d7a1e4c01949d6d3MD51TEXTILHA, GUSTAVO FREITAS.pdf.txtILHA, GUSTAVO FREITAS.pdf.txtExtracted texttext/plain73477http://repositorio.ufsm.br/bitstream/1/10101/2/ILHA%2c%20GUSTAVO%20FREITAS.pdf.txtcc298c9abdb0aadbca34394130a9c999MD52THUMBNAILILHA, GUSTAVO FREITAS.pdf.jpgILHA, GUSTAVO FREITAS.pdf.jpgIM Thumbnailimage/jpeg4664http://repositorio.ufsm.br/bitstream/1/10101/3/ILHA%2c%20GUSTAVO%20FREITAS.pdf.jpg53d28f4157c1dfbba4d36fbe4419432aMD531/101012023-05-30 10:42:29.42oai:repositorio.ufsm.br:1/10101Repositório Institucionalhttp://repositorio.ufsm.br/PUBhttp://repositorio.ufsm.br/oai/requestopendoar:39132023-05-30T13:42:29Manancial - Repositório Digital da UFSM - Universidade Federal de Santa Maria (UFSM)false
dc.title.por.fl_str_mv Caracterização do sistema calicreína-cinina durante o processo ovulatório de bovinos
dc.title.alternative.eng.fl_str_mv Characterization of kallikrein-kinin system during the ovulation process in bovine
title Caracterização do sistema calicreína-cinina durante o processo ovulatório de bovinos
spellingShingle Caracterização do sistema calicreína-cinina durante o processo ovulatório de bovinos
Ilha, Gustavo Freitas
Ovulação
Sistema calicreína-cinina
Bradicinina
Ovário
Bovino
Ovulation
Kallikrein-kinin system
Bradykinin
Ovary
Bovine
CNPQ::CIENCIAS AGRARIAS::MEDICINA VETERINARIA
title_short Caracterização do sistema calicreína-cinina durante o processo ovulatório de bovinos
title_full Caracterização do sistema calicreína-cinina durante o processo ovulatório de bovinos
title_fullStr Caracterização do sistema calicreína-cinina durante o processo ovulatório de bovinos
title_full_unstemmed Caracterização do sistema calicreína-cinina durante o processo ovulatório de bovinos
title_sort Caracterização do sistema calicreína-cinina durante o processo ovulatório de bovinos
author Ilha, Gustavo Freitas
author_facet Ilha, Gustavo Freitas
author_role author
dc.contributor.advisor1.fl_str_mv Oliveira, João Francisco Coelho de
dc.contributor.advisor1Lattes.fl_str_mv http://lattes.cnpq.br/1168979725730267
dc.contributor.referee1.fl_str_mv Mezzalira, Alceu
dc.contributor.referee1Lattes.fl_str_mv http://lattes.cnpq.br/2481625293830872
dc.contributor.referee2.fl_str_mv Ferreira, Juliano
dc.contributor.referee2Lattes.fl_str_mv http://lattes.cnpq.br/2694197910478313
dc.contributor.authorLattes.fl_str_mv http://lattes.cnpq.br/1907756288556494
dc.contributor.author.fl_str_mv Ilha, Gustavo Freitas
contributor_str_mv Oliveira, João Francisco Coelho de
Mezzalira, Alceu
Ferreira, Juliano
dc.subject.por.fl_str_mv Ovulação
Sistema calicreína-cinina
Bradicinina
Ovário
Bovino
topic Ovulação
Sistema calicreína-cinina
Bradicinina
Ovário
Bovino
Ovulation
Kallikrein-kinin system
Bradykinin
Ovary
Bovine
CNPQ::CIENCIAS AGRARIAS::MEDICINA VETERINARIA
dc.subject.eng.fl_str_mv Ovulation
Kallikrein-kinin system
Bradykinin
Ovary
Bovine
dc.subject.cnpq.fl_str_mv CNPQ::CIENCIAS AGRARIAS::MEDICINA VETERINARIA
description The kallikrein-kinin system (KKS) has been described as an important mediator of physiologic processes. Kallikreins use kininogen (KNG) as substrate to generate bradykinin, the principal active peptide of the KKS which acts through two types of receptors, the B1R and B2R. The objective of this study was to characterize some components of KKS in different compartments of ovary during the ovulation process in bovine. mRNA expression pattern of KNG, B1R and B2R was assessed in theca and granulosa cells and bradykinin concentration and kallikrein-like activity in follicular fluid of bovine peri-ovulatory follicles. In order to obtain a peri-ovulatory follicle (≥ 12mm), twenty-seven cows were submitted to estrus synchronization protocol and ovariectomized by colpotomy at 0, 3, 6, 12 or 24 hours after a GnRH-analog injection (gonadorelin; 100 μg, IM). Follicular fluid was aspirated for enzymatic assays and granulosa and theca cells were harvested for mRNA analysis. The mRNA expressions in follicular cells were evaluated by real-time RT-PCR and data represented as relative to housekeeping gene cyclophilin. Bradykinin concentration and Kallikrein-like activity was measured in follicular fluid by enzymatic immunoassay and selective substrate cleavage, respectively, and the absorbance measured using a plate reader. KNG mRNA expression was similar for both follicular cell types (P>0.05), while B2R expression in theca cells and B1R expression in theca and granulosa cells showed different profiles during peri-ovulatory period (P<0.05). Bradykinin concentration and kallikrein-like activity in follicular fluid were different (P<0.05) according the time during ovulation process. The results provide an important characterization of the presence and possible regulation of KKS during ovulation in bovine.
publishDate 2011
dc.date.issued.fl_str_mv 2011-02-25
dc.date.accessioned.fl_str_mv 2017-06-07
dc.date.available.fl_str_mv 2017-06-07
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dc.identifier.citation.fl_str_mv ILHA, Gustavo Freitas. Characterization of kallikrein-kinin system during the ovulation process in bovine. 2011. 40 f. Dissertação (Mestrado em Medicina Veterinária) - Universidade Federal de Santa Maria, Santa Maria, 2011.
dc.identifier.uri.fl_str_mv http://repositorio.ufsm.br/handle/1/10101
identifier_str_mv ILHA, Gustavo Freitas. Characterization of kallikrein-kinin system during the ovulation process in bovine. 2011. 40 f. Dissertação (Mestrado em Medicina Veterinária) - Universidade Federal de Santa Maria, Santa Maria, 2011.
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