Efeito da lectina Concanavalina A (CON A) sobre a ativação in vitro de folículos primordiais caprinos

Detalhes bibliográficos
Ano de defesa: 2013
Autor(a) principal: Portela, Antonia Moemia Lúcia Rodrigues
Orientador(a): Silva, José Roberto Viana
Banca de defesa: Não Informado pela instituição
Tipo de documento: Dissertação
Tipo de acesso: Acesso aberto
Idioma: por
Instituição de defesa: Não Informado pela instituição
Programa de Pós-Graduação: Não Informado pela instituição
Departamento: Não Informado pela instituição
País: Não Informado pela instituição
Palavras-chave em Português:
Link de acesso: http://www.repositorio.ufc.br/handle/riufc/15959
Resumo: The aim of this study were to evaluate the effect of different concentrations of Concanavalin A (Con A) and FSH on activation, and survival in vitro growth of goat primordial follicles, as well as investigating the effects of Con A and FSH on the expression of mRNA for PCNA, KL, c-Kit, BMP-15 and GDF-9. For this, ovarian cortex fragments were in vitro cultured for 1 to 6 days in α-MEM+ supplemented with 0, 5, 10, 20 or 40 μg/mL of Con A lectin (experiment 1). At the end of the growing period, the fragments of ovarian cortex were fixed for histology. Next, we evaluated the percentage of primordial follicles or under development. After determination of the concentration of Con A more efficient (10 μg/mL), fragments of ovarian cortex were cultured in situ for 6 days in α-MEM+ alone or supplemented with 50 ng/mL of FSH, 10 μg/mL of Con A or both (experiment 2). Tissues from the control uncultured or after culture in different treatments were fixed for histology or stored at -80ºC to assess the expression profile of mRNA for PCNA, KL, c-Kit, BMP-15 and GDF-9. The results of Experiment 1 demonstrated that after 6 days of culture, the percentage of developing follicles was significantly higher in samples cultured with α-MEM+ supplemented with 10 and 40 μg/mL Con A, when compared to the other treatments (P <0.05). In experiment 2, the presence of FSH, or both Con A promoted a reduction in the percentage of primordial follicles and increase developing follicles compared to control medium (α-MEM+). After 6 days of culture, the presence of FSH promoted an increase in mRNA levels for PCNA, but this effect was blocked by Con A. Moreover, the Con A increased mRNA levels for c-KIT compared to the control not grown, but Con A or Con A and FSH both reduced the levels of mRNA for KL. Furthermore, a reduction of BMP-15 mRNA was observed in follicles cultured in control medium, but the presence of FSH inhibited this reduction. Furthermore, Con A promoted an increase in levels of GDF-9 mRNA after in vitro culture. In conclusion, the Con A alone or in combination with FSH promote activation of primordial follicles goat after 6 days of in vitro culture and regulate the expression of mRNA for PCNA, KL, c-Kit, BMP-15 and GDF-9
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spelling Portela, Antonia Moemia Lúcia RodriguesSilva, José Roberto Viana2016-04-01T12:36:02Z2016-04-01T12:36:02Z2013PORTELA, A. M. L. R. Efeito da lectina Concanavalina A (CON A) sobre a ativação in vitro de folículos primordiais caprinos. 2013. 90 f. Dissertação (Mestrado em Biotecnologia) - Campus de Sobral, Universidade Federal do Ceará, Sobral, 2013.http://www.repositorio.ufc.br/handle/riufc/15959The aim of this study were to evaluate the effect of different concentrations of Concanavalin A (Con A) and FSH on activation, and survival in vitro growth of goat primordial follicles, as well as investigating the effects of Con A and FSH on the expression of mRNA for PCNA, KL, c-Kit, BMP-15 and GDF-9. For this, ovarian cortex fragments were in vitro cultured for 1 to 6 days in α-MEM+ supplemented with 0, 5, 10, 20 or 40 μg/mL of Con A lectin (experiment 1). At the end of the growing period, the fragments of ovarian cortex were fixed for histology. Next, we evaluated the percentage of primordial follicles or under development. After determination of the concentration of Con A more efficient (10 μg/mL), fragments of ovarian cortex were cultured in situ for 6 days in α-MEM+ alone or supplemented with 50 ng/mL of FSH, 10 μg/mL of Con A or both (experiment 2). Tissues from the control uncultured or after culture in different treatments were fixed for histology or stored at -80ºC to assess the expression profile of mRNA for PCNA, KL, c-Kit, BMP-15 and GDF-9. The results of Experiment 1 demonstrated that after 6 days of culture, the percentage of developing follicles was significantly higher in samples cultured with α-MEM+ supplemented with 10 and 40 μg/mL Con A, when compared to the other treatments (P <0.05). In experiment 2, the presence of FSH, or both Con A promoted a reduction in the percentage of primordial follicles and increase developing follicles compared to control medium (α-MEM+). After 6 days of culture, the presence of FSH promoted an increase in mRNA levels for PCNA, but this effect was blocked by Con A. Moreover, the Con A increased mRNA levels for c-KIT compared to the control not grown, but Con A or Con A and FSH both reduced the levels of mRNA for KL. Furthermore, a reduction of BMP-15 mRNA was observed in follicles cultured in control medium, but the presence of FSH inhibited this reduction. Furthermore, Con A promoted an increase in levels of GDF-9 mRNA after in vitro culture. In conclusion, the Con A alone or in combination with FSH promote activation of primordial follicles goat after 6 days of in vitro culture and regulate the expression of mRNA for PCNA, KL, c-Kit, BMP-15 and GDF-9Os objetivos deste estudo foram avaliar o efeito de diferentes concentrações de Concanavalina A (Con A) e do FSH sobre a ativação, sobrevivência e crescimento in vitro de folículos primordiais caprinos, bem como investigar os efeitos da Con A e do FSH sobre a expressão de RNAm para PCNA, KL, c-Kit, BMP-15 e GDF-9. Para isto, fragmentos de córtex ovariano foram cultivados in vitro por 1 ou 6 dias em α-MEM+ suplementado com 0, 5, 10, 20 ou 40 μg/mL da lectina Con A (experimento 1). No final do período de cultivo, os fragmentos de córtex ovariano foram fixados para histologia clássica. Em seguida, avaliou-se a percentagem de folículos primordiais ou em desenvolvimento. Após a determinação da concentração de Con A mais eficiente (10 μg/mL), os fragmentos do córtex ovariano foram cultivados in situ por 6 dias em α-MEM+ sozinho ou suplementado com 50 ng/mL de FSH, 10 μg/mL de Con A ou ambos (experimento 2). Os tecidos provenientes do controle não cultivado ou após cultivo nos diferentes tratamentos foram fixados para histologia clássica ou armazenados a -800C para avaliar o perfil de expressão de RNAs mensageiros para PCNA, KL, c-Kit, BMP-15 e GDF-9. Os resultados do experimento 1 demonstraram que após 6 dias de cultivo, a percentagem de folículos em desenvolvimento foi significativamente maior nos fragmentos cultivados com α-MEM+ suplementado com 10 e 40 μg/mL de Con A, quando comparado aos demais tratamentos (P <0,05). No experimento 2, a presença de FSH, Con A ou ambos promoveram uma redução na percentagem de folículos primordiais e aumento de folículos em desenvolvimento em relação ao meio controle (α-MEM+). Após 6 dias de cultivo, a presença de FSH promoveu um aumento nos níveis de RNAm para PCNA, mas este efeito foi bloqueado pela Con A. Por outro lado, a Con A aumentou os níveis de RNAm para c-kit em comparação ao controle não cultivado, mas Con-A ou ambos Con A e FSH reduziram os níveis de RNAm para KL. Além disso, uma redução dos RNAm para BMP-15 foi observado em folículos cultivados no meio controle, mas a presença de FSH inibiu esta redução. Além disso, a Con A promoveu um aumento nos níveis de RNAm para GDF-9 após cultivo in vitro. Em conclusão, a Con A sozinha ou em combinação com FSH promovem a ativação de folículos primordiais caprinos após 6 dias de cultivo in vitro e regulam a expressão de RNAm para PCNA, KL, c-Kit, BMP-15 e GDF-9.FSH-Hormônio FoliculoestimulanteConcanavalinaEfeito da lectina Concanavalina A (CON A) sobre a ativação in vitro de folículos primordiais caprinosEffect OF Concanavalin A (Con A) on the activaction in vitro goat primordial follicleinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/masterThesisporreponame:Repositório Institucional da Universidade Federal do Ceará (UFC)instname:Universidade Federal do Ceará (UFC)instacron:UFCinfo:eu-repo/semantics/openAccessORIGINAL2013_dis_amlrportela.pdf2013_dis_amlrportela.pdfapplication/pdf1263988http://repositorio.ufc.br/bitstream/riufc/15959/1/2013_dis_amlrportela.pdf5ba44fa2ebbd589c7c4c8cfd41dddd43MD51LICENSElicense.txtlicense.txttext/plain; charset=utf-81786http://repositorio.ufc.br/bitstream/riufc/15959/2/license.txt8c4401d3d14722a7ca2d07c782a1aab3MD52riufc/159592021-10-20 15:04:53.069oai:repositorio.ufc.br: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Repositório InstitucionalPUBhttp://www.repositorio.ufc.br/ri-oai/requestbu@ufc.br || repositorio@ufc.bropendoar:2021-10-20T18:04:53Repositório Institucional da Universidade Federal do Ceará (UFC) - Universidade Federal do Ceará (UFC)false
dc.title.pt_BR.fl_str_mv Efeito da lectina Concanavalina A (CON A) sobre a ativação in vitro de folículos primordiais caprinos
dc.title.en.pt_BR.fl_str_mv Effect OF Concanavalin A (Con A) on the activaction in vitro goat primordial follicle
title Efeito da lectina Concanavalina A (CON A) sobre a ativação in vitro de folículos primordiais caprinos
spellingShingle Efeito da lectina Concanavalina A (CON A) sobre a ativação in vitro de folículos primordiais caprinos
Portela, Antonia Moemia Lúcia Rodrigues
FSH-Hormônio Foliculoestimulante
Concanavalina
title_short Efeito da lectina Concanavalina A (CON A) sobre a ativação in vitro de folículos primordiais caprinos
title_full Efeito da lectina Concanavalina A (CON A) sobre a ativação in vitro de folículos primordiais caprinos
title_fullStr Efeito da lectina Concanavalina A (CON A) sobre a ativação in vitro de folículos primordiais caprinos
title_full_unstemmed Efeito da lectina Concanavalina A (CON A) sobre a ativação in vitro de folículos primordiais caprinos
title_sort Efeito da lectina Concanavalina A (CON A) sobre a ativação in vitro de folículos primordiais caprinos
author Portela, Antonia Moemia Lúcia Rodrigues
author_facet Portela, Antonia Moemia Lúcia Rodrigues
author_role author
dc.contributor.author.fl_str_mv Portela, Antonia Moemia Lúcia Rodrigues
dc.contributor.advisor1.fl_str_mv Silva, José Roberto Viana
contributor_str_mv Silva, José Roberto Viana
dc.subject.por.fl_str_mv FSH-Hormônio Foliculoestimulante
Concanavalina
topic FSH-Hormônio Foliculoestimulante
Concanavalina
description The aim of this study were to evaluate the effect of different concentrations of Concanavalin A (Con A) and FSH on activation, and survival in vitro growth of goat primordial follicles, as well as investigating the effects of Con A and FSH on the expression of mRNA for PCNA, KL, c-Kit, BMP-15 and GDF-9. For this, ovarian cortex fragments were in vitro cultured for 1 to 6 days in α-MEM+ supplemented with 0, 5, 10, 20 or 40 μg/mL of Con A lectin (experiment 1). At the end of the growing period, the fragments of ovarian cortex were fixed for histology. Next, we evaluated the percentage of primordial follicles or under development. After determination of the concentration of Con A more efficient (10 μg/mL), fragments of ovarian cortex were cultured in situ for 6 days in α-MEM+ alone or supplemented with 50 ng/mL of FSH, 10 μg/mL of Con A or both (experiment 2). Tissues from the control uncultured or after culture in different treatments were fixed for histology or stored at -80ºC to assess the expression profile of mRNA for PCNA, KL, c-Kit, BMP-15 and GDF-9. The results of Experiment 1 demonstrated that after 6 days of culture, the percentage of developing follicles was significantly higher in samples cultured with α-MEM+ supplemented with 10 and 40 μg/mL Con A, when compared to the other treatments (P <0.05). In experiment 2, the presence of FSH, or both Con A promoted a reduction in the percentage of primordial follicles and increase developing follicles compared to control medium (α-MEM+). After 6 days of culture, the presence of FSH promoted an increase in mRNA levels for PCNA, but this effect was blocked by Con A. Moreover, the Con A increased mRNA levels for c-KIT compared to the control not grown, but Con A or Con A and FSH both reduced the levels of mRNA for KL. Furthermore, a reduction of BMP-15 mRNA was observed in follicles cultured in control medium, but the presence of FSH inhibited this reduction. Furthermore, Con A promoted an increase in levels of GDF-9 mRNA after in vitro culture. In conclusion, the Con A alone or in combination with FSH promote activation of primordial follicles goat after 6 days of in vitro culture and regulate the expression of mRNA for PCNA, KL, c-Kit, BMP-15 and GDF-9
publishDate 2013
dc.date.issued.fl_str_mv 2013
dc.date.accessioned.fl_str_mv 2016-04-01T12:36:02Z
dc.date.available.fl_str_mv 2016-04-01T12:36:02Z
dc.type.status.fl_str_mv info:eu-repo/semantics/publishedVersion
dc.type.driver.fl_str_mv info:eu-repo/semantics/masterThesis
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dc.identifier.citation.fl_str_mv PORTELA, A. M. L. R. Efeito da lectina Concanavalina A (CON A) sobre a ativação in vitro de folículos primordiais caprinos. 2013. 90 f. Dissertação (Mestrado em Biotecnologia) - Campus de Sobral, Universidade Federal do Ceará, Sobral, 2013.
dc.identifier.uri.fl_str_mv http://www.repositorio.ufc.br/handle/riufc/15959
identifier_str_mv PORTELA, A. M. L. R. Efeito da lectina Concanavalina A (CON A) sobre a ativação in vitro de folículos primordiais caprinos. 2013. 90 f. Dissertação (Mestrado em Biotecnologia) - Campus de Sobral, Universidade Federal do Ceará, Sobral, 2013.
url http://www.repositorio.ufc.br/handle/riufc/15959
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