Regulation of gene expression during Drosophila oocyte development

Detalhes bibliográficos
Autor(a) principal: Matos, Ricardo
Data de Publicação: 2020
Tipo de documento: Dissertação
Idioma: eng
Título da fonte: Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)
Texto Completo: http://hdl.handle.net/10400.1/16694
Resumo: Human oocytes can stay dormant for several decades in meiotic arrest, before reactivation and maturation. Similar, Drosophila oocyte is mostly transcriptionally quiescent during prophase I-arrest, transiently reactivating gene expression just before it progresses into metaphase I. Our aim is to better understand such reactivation and its role for oocyte maturation and female fertility. This work is divided in 3 chapters. In chapter I we use an isolated loss of function allele of dkdm5 to characterize the function of this enzyme during oogenesis. The results obtained with the loss of function allele of dkdm5 are consistent, albeit stronger, with the phenotypes previously observed after RNAi depletion and hypomorphic allele of dkdm5. Yet, a qualitatively distinct phenotype has also been identified, suggesting a new function of dkdm5 during oocyte maturation. In chapter II we try to access chromatin quality and synaptonemal complex assembly throughout oocyte development. Proteins from chromatin remodelling complexes, as dkdm5, have been shown to disrupt the synaptonemal complex further leading to problems in meiotic progression. Here we observed that two core components of the pho repressive complex are required for female fertility. Further, we show that specific germline dSfmbt depletion leads to an abnormal increase of Corolla and defective synaptonemal complex morphology in the oocyte chromatin throughout development. Chapter III is an independent chapter where we reveal Nine Teen Complex Protein Salsa as being particularly rate limiting for efficient splicing of short proximal introns and dorsoventral patterning of the Drosophila egg. We observed that, upon specific germline depletion of Salsa, Gurken transcript is poorly spliced leading to an abnormal localization, subsequentially leading to defects in the eggshell dorsoventral patterning and female fertility. Further we show that ectopic Gurken expression can suppress the dorsal ventral patterning defects after Salsa depletion. Our work aims to mechanistically understand our observations.
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spelling Regulation of gene expression during Drosophila oocyte developmentDrosófilaOogénesedkdm5MeioseSynaptonemal complexDomínio/Área Científica::Ciências Médicas::Outras Ciências MédicasHuman oocytes can stay dormant for several decades in meiotic arrest, before reactivation and maturation. Similar, Drosophila oocyte is mostly transcriptionally quiescent during prophase I-arrest, transiently reactivating gene expression just before it progresses into metaphase I. Our aim is to better understand such reactivation and its role for oocyte maturation and female fertility. This work is divided in 3 chapters. In chapter I we use an isolated loss of function allele of dkdm5 to characterize the function of this enzyme during oogenesis. The results obtained with the loss of function allele of dkdm5 are consistent, albeit stronger, with the phenotypes previously observed after RNAi depletion and hypomorphic allele of dkdm5. Yet, a qualitatively distinct phenotype has also been identified, suggesting a new function of dkdm5 during oocyte maturation. In chapter II we try to access chromatin quality and synaptonemal complex assembly throughout oocyte development. Proteins from chromatin remodelling complexes, as dkdm5, have been shown to disrupt the synaptonemal complex further leading to problems in meiotic progression. Here we observed that two core components of the pho repressive complex are required for female fertility. Further, we show that specific germline dSfmbt depletion leads to an abnormal increase of Corolla and defective synaptonemal complex morphology in the oocyte chromatin throughout development. Chapter III is an independent chapter where we reveal Nine Teen Complex Protein Salsa as being particularly rate limiting for efficient splicing of short proximal introns and dorsoventral patterning of the Drosophila egg. We observed that, upon specific germline depletion of Salsa, Gurken transcript is poorly spliced leading to an abnormal localization, subsequentially leading to defects in the eggshell dorsoventral patterning and female fertility. Further we show that ectopic Gurken expression can suppress the dorsal ventral patterning defects after Salsa depletion. Our work aims to mechanistically understand our observations.Os oócitos humanos têm a habilidade de para o seu ciclo celular na meiose durante diversas décadas, num estado de dormência transcricional, antes de reativarem a sua transcrição e maturarem. Semelhante ao que ocorre em humanos, os oócitos de Drosófila também estão, na grande parte do seu desenvolvimento, com o seu ciclo celular parado na prófase I da meiose num estado de quiescência transcricional. Antes da progressão meiótica para a metáfase I, existe uma reativação transiente da transcrição no nucleou do oócito. Este trabalho está dividido em três capítulos. No primeiro capítulo, focamo-nos em utilizar um alelo isolado com a perda de função total de dkdm5 para tentar caracterizar a função desta enzina durante a oogénese. Os resultados obtidos são semelhantes aos observados anteriormente em experiências feitas através de depleção por RNAi ou pelo uso de um alelo hipomórfico. Contudo os fenótipos observados na perda da função total de dkdm5 são mais acentuados. Também, com este trabalho conseguimos desvendar alguns dos processos que são dependentes ou independentes da atividade de demetilase de dkdm5. Contudo, observamos um fenótipo qualitativamente distinto na cromatina do núcleo do oócito o que sugere uma nova função de dkdm5 durante a maturação do oócito. No segundo capítulo, avaliamos a qualidade da cromatina e a formação synaptonemal complex ao longo do desenvolvimento do oócito. Proteínas que integram complexos responsáveis pela remodelação da cromatina, como dkdm5, tem vindo a ser descritos como necessário para a correta formação do synaptonemal complex e subsequentemente uma correta progressão meiótica. Neste capítulo, mostramos que dois dos principais componentes de pho repressive complex, são necessários para a fertilidade feminina. Alem disso, mostramos também que após uma deleção especifica de dSfmbt na linha germinal, existe um aumento da expressão de Corolla e o aparecimento de morfologias defeituosas do SC durante o desenvolvimento do oócito. O terceiro capítulo é um capítulo independente. Neste capítulo nós descobrimos que a proteína Salsa do Nine Teen Complex protein como sendo particularmente limitante para a eficiência de splicing em intrões proximais curtos e padronização dorsoventral do ovo de Drosófila. Observamos que, após depleção especifica da linha germinal de Salsa que, o splincing do transcrito de Gurken não é feito corretamente levando a uma localização anormal desta proteína, subsequentemente levando a defeitos de padronização dorsoventral dos ovos e defeitos na fertilidade feminina. Contudo, mostramos que a expressão ectópica de Gurken pode suprimir os defeitos de padronização dorsoventral após a depleção de Salsa. O nosso objetivo reside em entender os mecanismos que estão por detrás das nossas observações.Martinho, Rui GonçaloSilva, RuiSapientiaMatos, Ricardo2022-07-01T00:30:18Z2020-12-022020-12-02T00:00:00Zinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/masterThesisapplication/pdfapplication/pdfapplication/pdfapplication/pdfapplication/pdfhttp://hdl.handle.net/10400.1/16694TID:202724891enginfo: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:RCAAP2023-07-24T10:28:42Zoai:sapientia.ualg.pt:10400.1/16694Portal AgregadorONGhttps://www.rcaap.pt/oai/openaireopendoar:71602024-03-19T20:06:47.126486Repositório 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 Regulation of gene expression during Drosophila oocyte development
title Regulation of gene expression during Drosophila oocyte development
spellingShingle Regulation of gene expression during Drosophila oocyte development
Matos, Ricardo
Drosófila
Oogénese
dkdm5
Meiose
Synaptonemal complex
Domínio/Área Científica::Ciências Médicas::Outras Ciências Médicas
title_short Regulation of gene expression during Drosophila oocyte development
title_full Regulation of gene expression during Drosophila oocyte development
title_fullStr Regulation of gene expression during Drosophila oocyte development
title_full_unstemmed Regulation of gene expression during Drosophila oocyte development
title_sort Regulation of gene expression during Drosophila oocyte development
author Matos, Ricardo
author_facet Matos, Ricardo
author_role author
dc.contributor.none.fl_str_mv Martinho, Rui Gonçalo
Silva, Rui
Sapientia
dc.contributor.author.fl_str_mv Matos, Ricardo
dc.subject.por.fl_str_mv Drosófila
Oogénese
dkdm5
Meiose
Synaptonemal complex
Domínio/Área Científica::Ciências Médicas::Outras Ciências Médicas
topic Drosófila
Oogénese
dkdm5
Meiose
Synaptonemal complex
Domínio/Área Científica::Ciências Médicas::Outras Ciências Médicas
description Human oocytes can stay dormant for several decades in meiotic arrest, before reactivation and maturation. Similar, Drosophila oocyte is mostly transcriptionally quiescent during prophase I-arrest, transiently reactivating gene expression just before it progresses into metaphase I. Our aim is to better understand such reactivation and its role for oocyte maturation and female fertility. This work is divided in 3 chapters. In chapter I we use an isolated loss of function allele of dkdm5 to characterize the function of this enzyme during oogenesis. The results obtained with the loss of function allele of dkdm5 are consistent, albeit stronger, with the phenotypes previously observed after RNAi depletion and hypomorphic allele of dkdm5. Yet, a qualitatively distinct phenotype has also been identified, suggesting a new function of dkdm5 during oocyte maturation. In chapter II we try to access chromatin quality and synaptonemal complex assembly throughout oocyte development. Proteins from chromatin remodelling complexes, as dkdm5, have been shown to disrupt the synaptonemal complex further leading to problems in meiotic progression. Here we observed that two core components of the pho repressive complex are required for female fertility. Further, we show that specific germline dSfmbt depletion leads to an abnormal increase of Corolla and defective synaptonemal complex morphology in the oocyte chromatin throughout development. Chapter III is an independent chapter where we reveal Nine Teen Complex Protein Salsa as being particularly rate limiting for efficient splicing of short proximal introns and dorsoventral patterning of the Drosophila egg. We observed that, upon specific germline depletion of Salsa, Gurken transcript is poorly spliced leading to an abnormal localization, subsequentially leading to defects in the eggshell dorsoventral patterning and female fertility. Further we show that ectopic Gurken expression can suppress the dorsal ventral patterning defects after Salsa depletion. Our work aims to mechanistically understand our observations.
publishDate 2020
dc.date.none.fl_str_mv 2020-12-02
2020-12-02T00:00:00Z
2022-07-01T00:30:18Z
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