Perfil da expressão de ISG15 no sangue periférico durante o período de reconhecimento materno da gestação em bovinos
Autor(a) principal: | |
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Data de Publicação: | 2018 |
Tipo de documento: | Dissertação |
Idioma: | por |
Título da fonte: | Repositório Institucional Manancial UFSM |
Texto Completo: | http://repositorio.ufsm.br/handle/1/14289 |
Resumo: | The maternal recognition of pregnancy (MRP) in ruminants is a complex event, involving several cellular mechanisms. It is at that moment that the highest embryonic mortality occurs in cattle. The MRP period is characterized by the secretion of interferon tau (IFNT) by the embryo. IFNT is produced and secreted by the trofloblast cells of the embryo, with greater intensity at 18 days of pregnancy. The main function of this protein is to keep the corpus luteum fully active, avoiding luteolysis. Direct action of IFNT on extrauterine tissues raises the expression of interferon-stimulated genes (ISGs). Among the ISGs stimulated by IFNT, the gene stimulated by interferon 15 (ISG15) stands out. Studies have demonstrated endocrine action of IFNT. An elevation in ISG expression levels in blood and luteal cells was observed shortly after signaling by IFNT at the start of gestation in ruminants. Peripheral blood polymorphonuclear cells (PMN) are more sensitive to the IFNT stimulus compared to peripheral blood mononuclear cells (PBMCs). The objectives of this study are: (1) to determine the earliest possible time to identify ISG15 expression in pregnant heifers; (2) establish the cell fraction to determine the early diagnosis of pregnancy with high accuracy in cut heifers; (3) demonstrate the expression profile of ISG15 on days 16 and 18 after artificial insemination (AI). Synchronization of the estrous cycle was performed in 28 heifers. The pregnancy diagnosis was performed on days 29 and 60 after AI and resulted in 10 pregnant heifers, 14 non-pregnant heifers, and 4 non-inseminated heifers (negative control group). Blood was collected from the coccygeal vein after AI (Days 16 and 18). Cellular fractions of PBMC and PMN were collected and stored at -80 ° C until further extraction of RNA, cDNA and qPCR, respectively. The relative expression of ISG15 differed between the PMN and PBMC cell fraction of pregnant heifers (p <0.0001). ISG15 expression in PMN cells on Day 16 differed from heifers and non-inseminated heifers (p <0.04), as well as on Day 18 after AI (p <0.01). However, ISG15 expression in PMN cells on Day 16 did not differ from pregnant heifers and non-pregnant heifers. In PBMC cells, ISG15 expression on Day 16 did not differ from pregnant, non-pregnant and non-inseminated heifers. However, on Day 18 after AI, there is a difference in ISG15 expression of pregnant and non-pregnant heifers (p <0.02). Expression of ISG15 in PMN cells on Days 16 and 18 pregnancy is greater 30 times than expression in PBMC. In addition, the expression of ISG15 in PMN can be used to identify embryonic signaling on Day 16 after insemination in beef heifers. Expression of ISG15 in PMN on Day 18 after AI resulted in 93% accuracy for the diagnosis of pregnancy. We suggest that both cellular fractions (PMN and PBMC) can be used as a tool to identify embryonic signaling on Day 18 of pregnancy in beef heifers. |
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2018-09-13T12:50:16Z2018-09-13T12:50:16Z2018-02-27http://repositorio.ufsm.br/handle/1/14289The maternal recognition of pregnancy (MRP) in ruminants is a complex event, involving several cellular mechanisms. It is at that moment that the highest embryonic mortality occurs in cattle. The MRP period is characterized by the secretion of interferon tau (IFNT) by the embryo. IFNT is produced and secreted by the trofloblast cells of the embryo, with greater intensity at 18 days of pregnancy. The main function of this protein is to keep the corpus luteum fully active, avoiding luteolysis. Direct action of IFNT on extrauterine tissues raises the expression of interferon-stimulated genes (ISGs). Among the ISGs stimulated by IFNT, the gene stimulated by interferon 15 (ISG15) stands out. Studies have demonstrated endocrine action of IFNT. An elevation in ISG expression levels in blood and luteal cells was observed shortly after signaling by IFNT at the start of gestation in ruminants. Peripheral blood polymorphonuclear cells (PMN) are more sensitive to the IFNT stimulus compared to peripheral blood mononuclear cells (PBMCs). The objectives of this study are: (1) to determine the earliest possible time to identify ISG15 expression in pregnant heifers; (2) establish the cell fraction to determine the early diagnosis of pregnancy with high accuracy in cut heifers; (3) demonstrate the expression profile of ISG15 on days 16 and 18 after artificial insemination (AI). Synchronization of the estrous cycle was performed in 28 heifers. The pregnancy diagnosis was performed on days 29 and 60 after AI and resulted in 10 pregnant heifers, 14 non-pregnant heifers, and 4 non-inseminated heifers (negative control group). Blood was collected from the coccygeal vein after AI (Days 16 and 18). Cellular fractions of PBMC and PMN were collected and stored at -80 ° C until further extraction of RNA, cDNA and qPCR, respectively. The relative expression of ISG15 differed between the PMN and PBMC cell fraction of pregnant heifers (p <0.0001). ISG15 expression in PMN cells on Day 16 differed from heifers and non-inseminated heifers (p <0.04), as well as on Day 18 after AI (p <0.01). However, ISG15 expression in PMN cells on Day 16 did not differ from pregnant heifers and non-pregnant heifers. In PBMC cells, ISG15 expression on Day 16 did not differ from pregnant, non-pregnant and non-inseminated heifers. However, on Day 18 after AI, there is a difference in ISG15 expression of pregnant and non-pregnant heifers (p <0.02). Expression of ISG15 in PMN cells on Days 16 and 18 pregnancy is greater 30 times than expression in PBMC. In addition, the expression of ISG15 in PMN can be used to identify embryonic signaling on Day 16 after insemination in beef heifers. Expression of ISG15 in PMN on Day 18 after AI resulted in 93% accuracy for the diagnosis of pregnancy. We suggest that both cellular fractions (PMN and PBMC) can be used as a tool to identify embryonic signaling on Day 18 of pregnancy in beef heifers.O período de reconhecimento materno da gestação (RMG) em ruminantes é um evento complexo, envolvendo diversos mecanismos celulares. É nesse momento que ocorre a maior mortalidade embrionária em bovinos. O período de RMG é caracterizado pela secreção de interferon tau (IFNT) pelas células do trofloblasto do embrião, com maior intensidade aos 18 dias de gestação. A principal função dessa proteína é manter o corpo lúteo em plena atividade, evitando a luteólise. A ação direta do IFNT em tecidos extrauterinos eleva a expressão de genes estimulados por interferon (ISGs destacando-se o gene estimulado por interferon 15 (ISG15). A elevação dos níveis da expressão de ISGs em células sanguíneas e luteais ocorrem logo após a sinalização por IFNT no início da gestação em ruminantes. As células polimorfonucleares do sangue periférico (PMN) são mais sensíveis ao estimulo do IFNT comparadas às células mononucleares do sangue periférico (PBMC). Os objetivos deste estudo são: (1) determinar o tempo mais precoce possível para identificar a expressão de ISG15 em novilhas de corte gestantes; (2) estabelecer a fração celular sanguínea para determinar o diagnóstico precoce de gestação com alta acurácia em novilhas de corte; e, (3) demonstrar o perfil da expressão de ISG15 nos dias 16 e 18 após a inseminação artificial (IA). A sincronização do ciclo estral foi realizada em 28 novilhas. O diagnóstico de gestação foi realizado nos dias 29 e 60 após IA e resultou em 10 novilhas gestantes, 14 novilhas não gestantes, e 4 novilhas não inseminadas (grupo controle negativo). O sangue foi coletado da veia coccígea após a IA (dias 16 e 18). As frações celulares de PBMC e PMN foram coletadas e armazenadas a -80ºC até a posterior manipulação de RNA, cDNA e qPCR. A expressão relativa de ISG15 diferiu entre a fração celular de PMN e PBMC de novilhas gestantes (p < 0,0001). A expressão do ISG15 em células PMN no dia 16 diferiu entre novilhas gestantes e novilhas não inseminadas (p < 0,04), bem como no dia 18 após IA (p < 0,01). No entanto, a expressão do ISG15 em células PMN no dia 16 não diferiu entre novilhas gestantes e novilhas não gestantes. Em PBMC, a expressão do ISG15 no dia 16 não diferiu entre as novilhas gestantes, não gestantes e não inseminadas. No entanto, no dia 18 após AI, há uma diferença na expressão de ISG15 entre novilhas gestantes e não gestantes (p < 0,02). A expressão de ISG15 nas células PMN aos 16 e 18 de gestação é ao menos 30 vezes maior do que a expressão em PBMC. Além disso, a expressão de ISG15 em PMN pode ser usada para identificação da sinalização embrionária aos 16 e 18 dias após a inseminação em novilhas de corte. A expressão de ISG15 em PMN no dia 18 após a IA resultou em 93% de acurácia para o diagnóstico de gestação. Sugerimos que ambas as frações celulares (PMN e PBMC) podem ser utilizadas como ferramentas para identificar a sinalização embrionária aos 18 dias de gestação em novilhas de corte.porUniversidade Federal de Santa MariaCentro de Ciências RuraisPrograma de Pós-Graduação em Medicina VeterináriaUFSMBrasilMedicina VeterináriaAttribution-NonCommercial-NoDerivatives 4.0 Internationalhttp://creativecommons.org/licenses/by-nc-nd/4.0/info:eu-repo/semantics/openAccessISG15Sinalização embrionáriaCélulas polimorfonucleares do sangue periféricoDiagnóstico de gestaçãoNovilhas de corteEmbryonic signalingPeripheral blood polymorphonuclear cellsDiagnosis of gestationBeef heifersCNPQ::CIENCIAS AGRARIAS::MEDICINA VETERINARIAPerfil da expressão de ISG15 no sangue periférico durante o período de reconhecimento materno da gestação em bovinosISG15 expression profile in peripheral blood during the maternal recognitionof pregnancy in cattleinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/masterThesisAntoniazzi, Alfredo Quiteshttp://lattes.cnpq.br/7258679373419746Mesquita, Fernando Silveirahttp://lattes.cnpq.br/3338405134728553Sudano, Mateus Joséhttp://lattes.cnpq.br/1945811068525845http://lattes.cnpq.br/5167001497034003Machado, Fabrício Amadori500500000007600a338fc55-084a-47a0-8cef-889209277d3d02a6aa55-1e26-42fb-a107-24e875a5a4274cd5e569-cc1a-47c5-8ad0-c8161879aeddcf2b44b6-6ae3-480f-bd1c-c9fc211bcc65reponame:Repositório Institucional Manancial UFSMinstname:Universidade Federal de Santa Maria (UFSM)instacron:UFSMCC-LICENSElicense_rdflicense_rdfapplication/rdf+xml; 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dc.title.por.fl_str_mv |
Perfil da expressão de ISG15 no sangue periférico durante o período de reconhecimento materno da gestação em bovinos |
dc.title.alternative.eng.fl_str_mv |
ISG15 expression profile in peripheral blood during the maternal recognitionof pregnancy in cattle |
title |
Perfil da expressão de ISG15 no sangue periférico durante o período de reconhecimento materno da gestação em bovinos |
spellingShingle |
Perfil da expressão de ISG15 no sangue periférico durante o período de reconhecimento materno da gestação em bovinos Machado, Fabrício Amadori ISG15 Sinalização embrionária Células polimorfonucleares do sangue periférico Diagnóstico de gestação Novilhas de corte Embryonic signaling Peripheral blood polymorphonuclear cells Diagnosis of gestation Beef heifers CNPQ::CIENCIAS AGRARIAS::MEDICINA VETERINARIA |
title_short |
Perfil da expressão de ISG15 no sangue periférico durante o período de reconhecimento materno da gestação em bovinos |
title_full |
Perfil da expressão de ISG15 no sangue periférico durante o período de reconhecimento materno da gestação em bovinos |
title_fullStr |
Perfil da expressão de ISG15 no sangue periférico durante o período de reconhecimento materno da gestação em bovinos |
title_full_unstemmed |
Perfil da expressão de ISG15 no sangue periférico durante o período de reconhecimento materno da gestação em bovinos |
title_sort |
Perfil da expressão de ISG15 no sangue periférico durante o período de reconhecimento materno da gestação em bovinos |
author |
Machado, Fabrício Amadori |
author_facet |
Machado, Fabrício Amadori |
author_role |
author |
dc.contributor.advisor1.fl_str_mv |
Antoniazzi, Alfredo Quites |
dc.contributor.advisor1Lattes.fl_str_mv |
http://lattes.cnpq.br/7258679373419746 |
dc.contributor.referee1.fl_str_mv |
Mesquita, Fernando Silveira |
dc.contributor.referee1Lattes.fl_str_mv |
http://lattes.cnpq.br/3338405134728553 |
dc.contributor.referee2.fl_str_mv |
Sudano, Mateus José |
dc.contributor.referee2Lattes.fl_str_mv |
http://lattes.cnpq.br/1945811068525845 |
dc.contributor.authorLattes.fl_str_mv |
http://lattes.cnpq.br/5167001497034003 |
dc.contributor.author.fl_str_mv |
Machado, Fabrício Amadori |
contributor_str_mv |
Antoniazzi, Alfredo Quites Mesquita, Fernando Silveira Sudano, Mateus José |
dc.subject.por.fl_str_mv |
ISG15 Sinalização embrionária Células polimorfonucleares do sangue periférico Diagnóstico de gestação Novilhas de corte |
topic |
ISG15 Sinalização embrionária Células polimorfonucleares do sangue periférico Diagnóstico de gestação Novilhas de corte Embryonic signaling Peripheral blood polymorphonuclear cells Diagnosis of gestation Beef heifers CNPQ::CIENCIAS AGRARIAS::MEDICINA VETERINARIA |
dc.subject.eng.fl_str_mv |
Embryonic signaling Peripheral blood polymorphonuclear cells Diagnosis of gestation Beef heifers |
dc.subject.cnpq.fl_str_mv |
CNPQ::CIENCIAS AGRARIAS::MEDICINA VETERINARIA |
description |
The maternal recognition of pregnancy (MRP) in ruminants is a complex event, involving several cellular mechanisms. It is at that moment that the highest embryonic mortality occurs in cattle. The MRP period is characterized by the secretion of interferon tau (IFNT) by the embryo. IFNT is produced and secreted by the trofloblast cells of the embryo, with greater intensity at 18 days of pregnancy. The main function of this protein is to keep the corpus luteum fully active, avoiding luteolysis. Direct action of IFNT on extrauterine tissues raises the expression of interferon-stimulated genes (ISGs). Among the ISGs stimulated by IFNT, the gene stimulated by interferon 15 (ISG15) stands out. Studies have demonstrated endocrine action of IFNT. An elevation in ISG expression levels in blood and luteal cells was observed shortly after signaling by IFNT at the start of gestation in ruminants. Peripheral blood polymorphonuclear cells (PMN) are more sensitive to the IFNT stimulus compared to peripheral blood mononuclear cells (PBMCs). The objectives of this study are: (1) to determine the earliest possible time to identify ISG15 expression in pregnant heifers; (2) establish the cell fraction to determine the early diagnosis of pregnancy with high accuracy in cut heifers; (3) demonstrate the expression profile of ISG15 on days 16 and 18 after artificial insemination (AI). Synchronization of the estrous cycle was performed in 28 heifers. The pregnancy diagnosis was performed on days 29 and 60 after AI and resulted in 10 pregnant heifers, 14 non-pregnant heifers, and 4 non-inseminated heifers (negative control group). Blood was collected from the coccygeal vein after AI (Days 16 and 18). Cellular fractions of PBMC and PMN were collected and stored at -80 ° C until further extraction of RNA, cDNA and qPCR, respectively. The relative expression of ISG15 differed between the PMN and PBMC cell fraction of pregnant heifers (p <0.0001). ISG15 expression in PMN cells on Day 16 differed from heifers and non-inseminated heifers (p <0.04), as well as on Day 18 after AI (p <0.01). However, ISG15 expression in PMN cells on Day 16 did not differ from pregnant heifers and non-pregnant heifers. In PBMC cells, ISG15 expression on Day 16 did not differ from pregnant, non-pregnant and non-inseminated heifers. However, on Day 18 after AI, there is a difference in ISG15 expression of pregnant and non-pregnant heifers (p <0.02). Expression of ISG15 in PMN cells on Days 16 and 18 pregnancy is greater 30 times than expression in PBMC. In addition, the expression of ISG15 in PMN can be used to identify embryonic signaling on Day 16 after insemination in beef heifers. Expression of ISG15 in PMN on Day 18 after AI resulted in 93% accuracy for the diagnosis of pregnancy. We suggest that both cellular fractions (PMN and PBMC) can be used as a tool to identify embryonic signaling on Day 18 of pregnancy in beef heifers. |
publishDate |
2018 |
dc.date.accessioned.fl_str_mv |
2018-09-13T12:50:16Z |
dc.date.available.fl_str_mv |
2018-09-13T12:50:16Z |
dc.date.issued.fl_str_mv |
2018-02-27 |
dc.type.status.fl_str_mv |
info:eu-repo/semantics/publishedVersion |
dc.type.driver.fl_str_mv |
info:eu-repo/semantics/masterThesis |
format |
masterThesis |
status_str |
publishedVersion |
dc.identifier.uri.fl_str_mv |
http://repositorio.ufsm.br/handle/1/14289 |
url |
http://repositorio.ufsm.br/handle/1/14289 |
dc.language.iso.fl_str_mv |
por |
language |
por |
dc.relation.cnpq.fl_str_mv |
500500000007 |
dc.relation.confidence.fl_str_mv |
600 |
dc.relation.authority.fl_str_mv |
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dc.rights.driver.fl_str_mv |
Attribution-NonCommercial-NoDerivatives 4.0 International http://creativecommons.org/licenses/by-nc-nd/4.0/ info:eu-repo/semantics/openAccess |
rights_invalid_str_mv |
Attribution-NonCommercial-NoDerivatives 4.0 International http://creativecommons.org/licenses/by-nc-nd/4.0/ |
eu_rights_str_mv |
openAccess |
dc.publisher.none.fl_str_mv |
Universidade Federal de Santa Maria Centro de Ciências Rurais |
dc.publisher.program.fl_str_mv |
Programa de Pós-Graduação em Medicina Veterinária |
dc.publisher.initials.fl_str_mv |
UFSM |
dc.publisher.country.fl_str_mv |
Brasil |
dc.publisher.department.fl_str_mv |
Medicina Veterinária |
publisher.none.fl_str_mv |
Universidade Federal de Santa Maria Centro de Ciências Rurais |
dc.source.none.fl_str_mv |
reponame:Repositório Institucional Manancial 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 |
Repositório Institucional Manancial UFSM |
collection |
Repositório Institucional Manancial UFSM |
bitstream.url.fl_str_mv |
http://repositorio.ufsm.br/bitstream/1/14289/2/license_rdf http://repositorio.ufsm.br/bitstream/1/14289/3/license.txt http://repositorio.ufsm.br/bitstream/1/14289/1/DIS_PPGMV_2018_MACHADO_FABRICIO.pdf http://repositorio.ufsm.br/bitstream/1/14289/4/DIS_PPGMV_2018_MACHADO_FABRICIO.pdf.txt http://repositorio.ufsm.br/bitstream/1/14289/5/DIS_PPGMV_2018_MACHADO_FABRICIO.pdf.jpg |
bitstream.checksum.fl_str_mv |
c1efe8e24d7281448e873be30ea326ff 2f0571ecee68693bd5cd3f17c1e075df 85fcabb25f774a3a956a573e16d4a889 5c52a58595fcb9a87f45071d4a9af9c4 c03061a52c274d691048e0cd9680af67 |
bitstream.checksumAlgorithm.fl_str_mv |
MD5 MD5 MD5 MD5 MD5 |
repository.name.fl_str_mv |
Repositório Institucional Manancial UFSM - Universidade Federal de Santa Maria (UFSM) |
repository.mail.fl_str_mv |
ouvidoria@ufsm.br |
_version_ |
1808854706242453504 |