DHT and E2 as modulators of apoptotic signaling in rat Sertoli cells

Detalhes bibliográficos
Autor(a) principal: Simões, Vera Lúcia Silveira
Data de Publicação: 2012
Tipo de documento: Dissertação
Idioma: eng
Título da fonte: Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)
Texto Completo: http://hdl.handle.net/10400.6/1115
Resumo: Apoptosis is an important regulatory event in testicular homeostasis and optimization of sperm production. Sertoli cells (SCs) form the blood-testis barrier creating a special microenvironment where germ cells develop and are under strict hormonal control. Estrogens and androgens are known to play critical roles in SCs functioning, improving their in vitro survival by preventing apoptotic progression. Herein, the influence of 17β-estradiol (E2) and 5α-dihydrotestosterone (DHT) on the apoptotic signaling pathways of immature rat cultured SCs was studied. For that purpose key points of the apoptotic pathway that interact with the mitochondria were chosen and the mRNA expression and/or protein levels of several apoptotic markers such as p53, the pro-apoptotic Bcl2 family member Bax, the apoptosis-inducing factor (AIF) and caspase-3 and 9 were evaluated. Caspase-3 activity was also evaluated as an endpoint marker of apoptosis. E2 and DHT down-regulated the mRNA transcript levels of p53, Bax, caspase-9 and caspase-3. The protein levels of AIF were reduced after DHT treatment while E2-treated cells decreased the cleaved caspase-9 protein levels. The apoptotic endpoint Caspase-3 activity presented highly decreased levels after hormonal treatment. Taken together, these results showed that E2 and DHT act as apoptotic signaling modulators in in vitro immature rat SCs suggesting that androgens and estrogens may be capable of modulating independent pathways of the apoptotic event by regulating different pro-apoptotic factors.
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spelling DHT and E2 as modulators of apoptotic signaling in rat Sertoli cellsCélulas de SertoliApoptoseAndrogéniosEstrogéniosCaspasesApoptosis is an important regulatory event in testicular homeostasis and optimization of sperm production. Sertoli cells (SCs) form the blood-testis barrier creating a special microenvironment where germ cells develop and are under strict hormonal control. Estrogens and androgens are known to play critical roles in SCs functioning, improving their in vitro survival by preventing apoptotic progression. Herein, the influence of 17β-estradiol (E2) and 5α-dihydrotestosterone (DHT) on the apoptotic signaling pathways of immature rat cultured SCs was studied. For that purpose key points of the apoptotic pathway that interact with the mitochondria were chosen and the mRNA expression and/or protein levels of several apoptotic markers such as p53, the pro-apoptotic Bcl2 family member Bax, the apoptosis-inducing factor (AIF) and caspase-3 and 9 were evaluated. Caspase-3 activity was also evaluated as an endpoint marker of apoptosis. E2 and DHT down-regulated the mRNA transcript levels of p53, Bax, caspase-9 and caspase-3. The protein levels of AIF were reduced after DHT treatment while E2-treated cells decreased the cleaved caspase-9 protein levels. The apoptotic endpoint Caspase-3 activity presented highly decreased levels after hormonal treatment. Taken together, these results showed that E2 and DHT act as apoptotic signaling modulators in in vitro immature rat SCs suggesting that androgens and estrogens may be capable of modulating independent pathways of the apoptotic event by regulating different pro-apoptotic factors.A apoptose é um evento regulador importante na homeostase testicular e na otimização da produção de espermatozoides. As células de Sertoli formam a barreira hematotesticular, criando um microambiente especial onde as células germinativas se desenvolvem e estão sob controle hormonal estrito. De facto, os estrogénios e os androgénios são conhecidos por desempenharem um papel importante no funcionamento das células de Sertoli, melhorando a sua sobrevivência in vitro e impedindo a progressão da apoptose. Neste trabalho estudou-se a influência do 17β-estradiol (E2) e da 5αdihidrotestosterona (DHT) nas vias de sinalização apoptótica em culturas de células de Sertoli de ratos imaturos. Para isso, foram selecionados pontos chave da via apoptótica que interagem com a mitocôndria e foram avaliados os níveis de expressão de mRNA e/ou dos níveis de proteína de vários desses marcadores apoptóticos, tais como o p53, o membro próapoptótico da família Bcl-2 designado por Bax, o factor de indução de apoptose (AIF) e as caspase-9 e caspase-3. A actividade da caspase-3 foi também avaliada como um marcador da fase final da apoptose. Pudemos verificar que tanto o E2 como a DHT diminuíram os níveis de transcritos de mRNA do p53, Bax, caspase-9 e caspase-3. Os níveis proteicos de AIF foram também reduzidos após o tratamento com DHT, enquanto que nas células tratadas com E2, se verificou uma diminuição nos níveis proteicos da caspase-9 clivada. A actividade da caspase-3 apresentou uma acentuada diminuição após o tratamento hormonal quer com E2, quer com DHT. Em conjunto, estes resultados demonstram que o E2 e a DHT actuam como moduladores in vitro da sinalização apoptótica em células de Sertoli de ratos imaturos, sugerindo que, nessas células, os androgénios e os estrogénios podem ser capazes de modular vias independentes da apoptose, uma vez que parecem regular diferencialmente a expressão de factores pró-apoptóticos distintos.Universidade da Beira InteriorOliveira, Carlos Pedro FontesuBibliorumSimões, Vera Lúcia Silveira2013-03-25T14:47:31Z2012-062012-06-01T00:00:00Zinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/masterThesisapplication/pdfhttp://hdl.handle.net/10400.6/1115enginfo:eu-repo/semantics/openAccessreponame:Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)instname:Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informaçãoinstacron:RCAAP2023-12-15T09:36:36Zoai:ubibliorum.ubi.pt:10400.6/1115Portal AgregadorONGhttps://www.rcaap.pt/oai/openaireopendoar:71602024-03-20T00:43:02.272562Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) - Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informaçãofalse
dc.title.none.fl_str_mv DHT and E2 as modulators of apoptotic signaling in rat Sertoli cells
title DHT and E2 as modulators of apoptotic signaling in rat Sertoli cells
spellingShingle DHT and E2 as modulators of apoptotic signaling in rat Sertoli cells
Simões, Vera Lúcia Silveira
Células de Sertoli
Apoptose
Androgénios
Estrogénios
Caspases
title_short DHT and E2 as modulators of apoptotic signaling in rat Sertoli cells
title_full DHT and E2 as modulators of apoptotic signaling in rat Sertoli cells
title_fullStr DHT and E2 as modulators of apoptotic signaling in rat Sertoli cells
title_full_unstemmed DHT and E2 as modulators of apoptotic signaling in rat Sertoli cells
title_sort DHT and E2 as modulators of apoptotic signaling in rat Sertoli cells
author Simões, Vera Lúcia Silveira
author_facet Simões, Vera Lúcia Silveira
author_role author
dc.contributor.none.fl_str_mv Oliveira, Carlos Pedro Fontes
uBibliorum
dc.contributor.author.fl_str_mv Simões, Vera Lúcia Silveira
dc.subject.por.fl_str_mv Células de Sertoli
Apoptose
Androgénios
Estrogénios
Caspases
topic Células de Sertoli
Apoptose
Androgénios
Estrogénios
Caspases
description Apoptosis is an important regulatory event in testicular homeostasis and optimization of sperm production. Sertoli cells (SCs) form the blood-testis barrier creating a special microenvironment where germ cells develop and are under strict hormonal control. Estrogens and androgens are known to play critical roles in SCs functioning, improving their in vitro survival by preventing apoptotic progression. Herein, the influence of 17β-estradiol (E2) and 5α-dihydrotestosterone (DHT) on the apoptotic signaling pathways of immature rat cultured SCs was studied. For that purpose key points of the apoptotic pathway that interact with the mitochondria were chosen and the mRNA expression and/or protein levels of several apoptotic markers such as p53, the pro-apoptotic Bcl2 family member Bax, the apoptosis-inducing factor (AIF) and caspase-3 and 9 were evaluated. Caspase-3 activity was also evaluated as an endpoint marker of apoptosis. E2 and DHT down-regulated the mRNA transcript levels of p53, Bax, caspase-9 and caspase-3. The protein levels of AIF were reduced after DHT treatment while E2-treated cells decreased the cleaved caspase-9 protein levels. The apoptotic endpoint Caspase-3 activity presented highly decreased levels after hormonal treatment. Taken together, these results showed that E2 and DHT act as apoptotic signaling modulators in in vitro immature rat SCs suggesting that androgens and estrogens may be capable of modulating independent pathways of the apoptotic event by regulating different pro-apoptotic factors.
publishDate 2012
dc.date.none.fl_str_mv 2012-06
2012-06-01T00:00:00Z
2013-03-25T14:47:31Z
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.uri.fl_str_mv http://hdl.handle.net/10400.6/1115
url http://hdl.handle.net/10400.6/1115
dc.language.iso.fl_str_mv eng
language eng
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dc.format.none.fl_str_mv application/pdf
dc.publisher.none.fl_str_mv Universidade da Beira Interior
publisher.none.fl_str_mv Universidade da Beira Interior
dc.source.none.fl_str_mv reponame:Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)
instname:Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação
instacron:RCAAP
instname_str Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação
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collection Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)
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