Caracterização do locus Fhos e sua participação na morte celular programada do intestino médio larval de Drosophila melanogaster

Detalhes bibliográficos
Ano de defesa: 2014
Autor(a) principal: BENZE JÚNIOR, Roosevelt Antonio lattes
Orientador(a): SIMON, Claudio Roberto lattes
Banca de defesa: Não Informado pela instituição
Tipo de documento: Dissertação
Tipo de acesso: Acesso aberto
Idioma: por
Instituição de defesa: Universidade Federal do Triângulo Mineiro
Programa de Pós-Graduação: Programa de Pós-Graduação em Ciências Fisiológicas
Departamento: Instituto de Ciências da Saúde - ICS::Curso de Medicina
País: Brasil
Palavras-chave em Português:
Palavras-chave em Inglês:
Área do conhecimento CNPq:
Link de acesso: http://bdtd.uftm.edu.br/handle/tede/576
Resumo: Throughout metazoans physiological or programmed cell death (PCD), in its different fashion, apoptotic, autophagic and necrotic, plays a central role in maintaining homeostasis. Several diseases are associated with the misregulation of PCD, such as degenerative and autoimmune diseases as well as cancer. Several components of the PCD machinery were evolutionarily conserved even in such distinct organisms as insect and humans. In addition to these similarities, the variety of experimental tools available make Drosophila a powerfull for the characterization of new cell death-related genetic functions. Our group has used the PCD of larval salivary glands as an experimental model to study physiological cell death processes during Drosophila’s development. The PCD in larval salivary glands is regulated by ecdysone (steroid Hormone). Despite the extensive knowledge of the transcriptional cascade triggered by ecdysone, how, this systemic signal present in the hemolymph, ultimately promotes tissue and stage specific responses it is still unclear. To answer this question, we have carried out a genetic screen to identify new genes associated with regulation and execution of salivary gland PCD. In such screen we have identified a mutation mapped on the the Fhos locus which cause a cell death defective phenotype, the persistence of salivary glands. The genetic and functional characterization of the locus have shown that its function is essential for the proper PCD of salivary glands. The Fhos locus encodes proteins that share high similarity to the Formin family of proteins which have been shown to play a role in active cytoskeleton remodeling in distinct biological processes. In this work, we have performed the transcriptional analysis of one of the isoforms encoded by the Fhos locus, the Fhos-C isoform, that showed that this isoform is probably the fusion between Fhos and a flanking gene called “desert”. The trasncriptional analysis also revealed that the Fhos-C is diferentially expressed during development in lower levels. We have also started functional analysis of the Fhos locus using transgenic lines aiming to evaluate the potential function of it in the PCD of the larval midgut that is also eliminated by PCD during development. After determining the transcriptional profile of the biologicaly active isoform in the midgut (FhosB) we carried out overexpression and knockdown assays of this particular isoform. Taken together, our results reveal that the isoform-C is not active in the PCD during insect metamorphosis. Either, overexpression or knockdown of the isoform Fhos-B did not produced any relevant phenotype differently of the drastic one seen in salivary glands.
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spelling SIMON, Claudio Robertohttp://lattes.cnpq.br/5641601085787358ARAUJO, Lucas Anhezini dehttp://lattes.cnpq.br/7187546927822723http://lattes.cnpq.br/7100253147473031BENZE JÚNIOR, Roosevelt Antonio2018-06-08T17:45:24Z2014-06-16BENZE JÚNIOR, Roosevelt Antonio. Caracterização do locus Fhos e sua participação na morte celular programada do intestino médio larval de Drosophila melanogaster. 2014. 101f. Dissertação (Mestrado em Ciências Fisiológicas) - Programa de Pós-Graduação em Ciências Fisiológicas, Universidade Federal do Triângulo Mineiro, Uberaba, 2014.http://bdtd.uftm.edu.br/handle/tede/576Throughout metazoans physiological or programmed cell death (PCD), in its different fashion, apoptotic, autophagic and necrotic, plays a central role in maintaining homeostasis. Several diseases are associated with the misregulation of PCD, such as degenerative and autoimmune diseases as well as cancer. Several components of the PCD machinery were evolutionarily conserved even in such distinct organisms as insect and humans. In addition to these similarities, the variety of experimental tools available make Drosophila a powerfull for the characterization of new cell death-related genetic functions. Our group has used the PCD of larval salivary glands as an experimental model to study physiological cell death processes during Drosophila’s development. The PCD in larval salivary glands is regulated by ecdysone (steroid Hormone). Despite the extensive knowledge of the transcriptional cascade triggered by ecdysone, how, this systemic signal present in the hemolymph, ultimately promotes tissue and stage specific responses it is still unclear. To answer this question, we have carried out a genetic screen to identify new genes associated with regulation and execution of salivary gland PCD. In such screen we have identified a mutation mapped on the the Fhos locus which cause a cell death defective phenotype, the persistence of salivary glands. The genetic and functional characterization of the locus have shown that its function is essential for the proper PCD of salivary glands. The Fhos locus encodes proteins that share high similarity to the Formin family of proteins which have been shown to play a role in active cytoskeleton remodeling in distinct biological processes. In this work, we have performed the transcriptional analysis of one of the isoforms encoded by the Fhos locus, the Fhos-C isoform, that showed that this isoform is probably the fusion between Fhos and a flanking gene called “desert”. The trasncriptional analysis also revealed that the Fhos-C is diferentially expressed during development in lower levels. We have also started functional analysis of the Fhos locus using transgenic lines aiming to evaluate the potential function of it in the PCD of the larval midgut that is also eliminated by PCD during development. After determining the transcriptional profile of the biologicaly active isoform in the midgut (FhosB) we carried out overexpression and knockdown assays of this particular isoform. Taken together, our results reveal that the isoform-C is not active in the PCD during insect metamorphosis. Either, overexpression or knockdown of the isoform Fhos-B did not produced any relevant phenotype differently of the drastic one seen in salivary glands.Nos metazoários a morte celular fisiológica ou programada (MCP), nas suas diferentes formas, apoptótica, autofágica ou necrótica desempenha papel central na manutenção da homeostase. Várias doenças estão correlacionadas às disfunções da regulação da MCP, dentre elas podemos destacar doenças degenerativas, autoimunes e o câncer. Vários componentes da maquinaria da MCP foram conservados evolutivamente e apresentam grande semelhança mesmo entre organismos tão distintos quanto os insetos e seres humanos. Estas semelhanças, juntamente com a grande variedade de ferramentas experimentais disponíveis tornam a Drosophila melanogaster um modelo poderoso para a caracterização de novas funções gênicas relacionadas à MCP. Nosso grupo tem utilizado a MCP das glândulas salivares larvais como modelo experimental para o estudo de processos de morte celular fisiológicos durante o desenvolvimento de Drosophila. A MCP das glândulas salivares larvais é regulada pela ecdisona (hormônio esteróide). Embora a cascata de ativação transcricional induzida por ecdisona seja conhecida, em última instância, como esse sinal sistêmico (ecdisona), presente na hemolinfa do inseto, promove respostas estágio e tecido específicas ainda é pouco conhecido. Para responder essa pergunta, realizamos um “screen” genético com o intuito de identificar novos genes associados, à regulação ou execução da MCP de glândulas salivares. Neste “screening” identificamos uma mutação mapeada no locus Fhos que causa um fenótipo defectivo de MCP, a persistência de glândulas salivares larvais. A caracterização genética desse locus demonstrou que sua função é essencial para a ocorrência da MCP de glândulas salivares. O locus Fhos codifica proteínas com alta similaridade à família de proteínas forminas as quais participam do remodelamento dinâmico do citoesqueleto em diferentes processos biológicos. Neste trabalho, realizamos análises transcricionais e in silico de uma das isoformas codificadas pelo locus Fhos, a isoforma Fhos-C, as quais revelaram ser esta fruto de uma fusão gênica entre Fhos e um gene flanqueador denominado “desert”. A análise transcricional de Fhos-C revelou que este transcrito é diferencialmente expresso e geralmente em baixos níveis. Iniciamos análises funcionais do locus Fhos através da utilização de linhagens transgênicas na tentativa de avaliar a potencial função deste locus na MCP do intestino larval o qual e também eliminado por MCP durante o desenvolvimento pupal. Após determinar o perfil de transcrição da isoforma biologicamente ativa (Fhos-B) na MCP do intestino larval foram realizados ensaios de superexpressão e de knockdown desta isoforma. Em conjunto nossos dados mostram que a isoforma Fhos-C, parece não participar da MCP durante a metamorfose do inseto. A indução da expressão ectópica e o “knockdown” tecido específico da isoforma Fhos-B não produziu fenótipo evidente quanto aquele observado na MCP de glândulas salivares.Coordenação de Aperfeiçoamento de Pessoal de Nível SuperiorFundação de Amparo à Pesquisa do Estado de Minas Geraisapplication/pdfhttp://bdtd.uftm.edu.br/retrieve/3510/Dissert%20Roosevelt%20A%20Benze%20Junior.pdf.jpgporUniversidade Federal do Triângulo MineiroPrograma de Pós-Graduação em Ciências FisiológicasUFTMBrasilInstituto de Ciências da Saúde - ICS::Curso de MedicinaAdams M.D., Celniker S.E., Holt R.A., Evans C.A., Gocayne J.D., Amanatides P.G., Scherer S.E., Li P.W., Hoskins R.A., Galle R.F., George R.A., Lewis S.E., Richards S., Ashburner M., Henderson S.N., Sutton G.G., Wortman J.R., Yandell M.D., Zhang Q., Chen L.X., Brandon R.C., Rogers Y.H., Blazej R.G., Champe M., Pfeiffer B.D., Wan K.H., Doyle C., Baxter E.G., Helt G., Nelson C.R., Gabor L.G., Abril J.F., Agbayani A., An H.J., Andrews-Pfannkoch C., Baldwin D., Ballew R.M., Basu A., Baxendale J., Bayraktaroglu L., Beasley E.M., Beeson K.Y., Benos P.V., Berman B.P., Bhandari D., Bolshakov S., Borkova D., Botchan M.R., Bouck J., Brokstein P., Brottier P., Burtis K. 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dc.title.por.fl_str_mv Caracterização do locus Fhos e sua participação na morte celular programada do intestino médio larval de Drosophila melanogaster
dc.title.alternative.eng.fl_str_mv Characterization of the Fhos locus and its participation in the larval midgut cell death of Drosophila melanogaster
title Caracterização do locus Fhos e sua participação na morte celular programada do intestino médio larval de Drosophila melanogaster
spellingShingle Caracterização do locus Fhos e sua participação na morte celular programada do intestino médio larval de Drosophila melanogaster
BENZE JÚNIOR, Roosevelt Antonio
Drosophila Melanogaster.
Morte celular.
Autofagia
cell death
Drosophila Melanogaster.
Fisiologia
title_short Caracterização do locus Fhos e sua participação na morte celular programada do intestino médio larval de Drosophila melanogaster
title_full Caracterização do locus Fhos e sua participação na morte celular programada do intestino médio larval de Drosophila melanogaster
title_fullStr Caracterização do locus Fhos e sua participação na morte celular programada do intestino médio larval de Drosophila melanogaster
title_full_unstemmed Caracterização do locus Fhos e sua participação na morte celular programada do intestino médio larval de Drosophila melanogaster
title_sort Caracterização do locus Fhos e sua participação na morte celular programada do intestino médio larval de Drosophila melanogaster
author BENZE JÚNIOR, Roosevelt Antonio
author_facet BENZE JÚNIOR, Roosevelt Antonio
author_role author
dc.contributor.advisor1.fl_str_mv SIMON, Claudio Roberto
dc.contributor.advisor1Lattes.fl_str_mv http://lattes.cnpq.br/5641601085787358
dc.contributor.advisor-co1.fl_str_mv ARAUJO, Lucas Anhezini de
dc.contributor.advisor-co1Lattes.fl_str_mv http://lattes.cnpq.br/7187546927822723
dc.contributor.authorLattes.fl_str_mv http://lattes.cnpq.br/7100253147473031
dc.contributor.author.fl_str_mv BENZE JÚNIOR, Roosevelt Antonio
contributor_str_mv SIMON, Claudio Roberto
ARAUJO, Lucas Anhezini de
dc.subject.por.fl_str_mv Drosophila Melanogaster.
Morte celular.
Autofagia
topic Drosophila Melanogaster.
Morte celular.
Autofagia
cell death
Drosophila Melanogaster.
Fisiologia
dc.subject.eng.fl_str_mv cell death
Drosophila Melanogaster.
dc.subject.cnpq.fl_str_mv Fisiologia
description Throughout metazoans physiological or programmed cell death (PCD), in its different fashion, apoptotic, autophagic and necrotic, plays a central role in maintaining homeostasis. Several diseases are associated with the misregulation of PCD, such as degenerative and autoimmune diseases as well as cancer. Several components of the PCD machinery were evolutionarily conserved even in such distinct organisms as insect and humans. In addition to these similarities, the variety of experimental tools available make Drosophila a powerfull for the characterization of new cell death-related genetic functions. Our group has used the PCD of larval salivary glands as an experimental model to study physiological cell death processes during Drosophila’s development. The PCD in larval salivary glands is regulated by ecdysone (steroid Hormone). Despite the extensive knowledge of the transcriptional cascade triggered by ecdysone, how, this systemic signal present in the hemolymph, ultimately promotes tissue and stage specific responses it is still unclear. To answer this question, we have carried out a genetic screen to identify new genes associated with regulation and execution of salivary gland PCD. In such screen we have identified a mutation mapped on the the Fhos locus which cause a cell death defective phenotype, the persistence of salivary glands. The genetic and functional characterization of the locus have shown that its function is essential for the proper PCD of salivary glands. The Fhos locus encodes proteins that share high similarity to the Formin family of proteins which have been shown to play a role in active cytoskeleton remodeling in distinct biological processes. In this work, we have performed the transcriptional analysis of one of the isoforms encoded by the Fhos locus, the Fhos-C isoform, that showed that this isoform is probably the fusion between Fhos and a flanking gene called “desert”. The trasncriptional analysis also revealed that the Fhos-C is diferentially expressed during development in lower levels. We have also started functional analysis of the Fhos locus using transgenic lines aiming to evaluate the potential function of it in the PCD of the larval midgut that is also eliminated by PCD during development. After determining the transcriptional profile of the biologicaly active isoform in the midgut (FhosB) we carried out overexpression and knockdown assays of this particular isoform. Taken together, our results reveal that the isoform-C is not active in the PCD during insect metamorphosis. Either, overexpression or knockdown of the isoform Fhos-B did not produced any relevant phenotype differently of the drastic one seen in salivary glands.
publishDate 2014
dc.date.issued.fl_str_mv 2014-06-16
dc.date.accessioned.fl_str_mv 2018-06-08T17:45:24Z
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.citation.fl_str_mv BENZE JÚNIOR, Roosevelt Antonio. Caracterização do locus Fhos e sua participação na morte celular programada do intestino médio larval de Drosophila melanogaster. 2014. 101f. Dissertação (Mestrado em Ciências Fisiológicas) - Programa de Pós-Graduação em Ciências Fisiológicas, Universidade Federal do Triângulo Mineiro, Uberaba, 2014.
dc.identifier.uri.fl_str_mv http://bdtd.uftm.edu.br/handle/tede/576
identifier_str_mv BENZE JÚNIOR, Roosevelt Antonio. Caracterização do locus Fhos e sua participação na morte celular programada do intestino médio larval de Drosophila melanogaster. 2014. 101f. Dissertação (Mestrado em Ciências Fisiológicas) - Programa de Pós-Graduação em Ciências Fisiológicas, Universidade Federal do Triângulo Mineiro, Uberaba, 2014.
url http://bdtd.uftm.edu.br/handle/tede/576
dc.language.iso.fl_str_mv por
language por
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