Using cilia mutants to study left-right asymmetry in zebrafish

Detalhes bibliográficos
Autor(a) principal: Sampaio, Pedro Rafael Martins de Almeida
Data de Publicação: 2014
Tipo de documento: Dissertação
Idioma: eng
Título da fonte: Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)
Texto Completo: http://hdl.handle.net/10362/13155
Resumo: A thesis submitted in fulfillment of the requirements for the degree of the Masters in Molecular Genetics and Biomedicine
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spelling Using cilia mutants to study left-right asymmetry in zebrafishKupffer’s vesicleMotile ciliaKV flowLeft-right asymmetryDnah7DeltaD mutantA thesis submitted in fulfillment of the requirements for the degree of the Masters in Molecular Genetics and BiomedicineIn vertebrates, internal organs are positioned asymmetrically across the left-right (L-R) body axis. Events determining L-R asymmetry occur during embryogenesis, and are regulated by the coordinated action of genetic mechanisms. Embryonic motile cilia are essential in this process by generating a directional fluid flow inside the zebrafish organ of asymmetry, called Kupffer’s vesicle ﴾KV). A correct L-R formation is highly dependent on signaling pathways downstream of such flow, however detailed characterization of how its dynamics modulates these mechanisms is still lacking. In this project, fluid flow measurements were achieved by a non-invasive method, in four genetic backgrounds: Wild-type (WT), deltaD-/- mutants, Dnah7 morphants (MO) and control-MO embryos. Knockdown of Dnah7, a heavy chain inner axonemal dynein, renders cilia completely immotile and depletes the KV directional fluid flow, which we characterize here for the first time. By following the development of each embryo, we show that flow dynamics in the KV is already asymmetric and provides a very good prediction of organ laterality. Through novel experiments, we characterized a new population of motile cilia, an immotile population, a range of cilia beat frequencies and lengths, KV volumes and cilia numbers in live embryos. These data were crucial to perform fluid dynamics simulations, which suggested that the flow in embryos with 30 or more cilia reliably produces left situs; with fewer cilia, left situs is sometimes compromised through disruption of the dorsal anterior clustering of motile cilia. A rough estimate based upon the 30 cilium threshold and statistics of cilium number predicts 90% and 60% left situs in WT and deltaD-/- respectively, as observed experimentally. Cilia number and clustering are therefore critical to normal situs via robust asymmetric flow. Thus, our results support a model in which asymmetric flow forces registered in the KV pattern organ laterality in each embryo.Faculdade de Ciências e TecnologiaLopes, SusanaLudovice, AnaRUNSampaio, Pedro Rafael Martins de Almeida2014-09-30T10:37:31Z20142014-01-01T00:00:00Zinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/masterThesisapplication/pdfhttp://hdl.handle.net/10362/13155enginfo: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:RCAAP2024-03-11T03:47:56Zoai:run.unl.pt:10362/13155Portal AgregadorONGhttps://www.rcaap.pt/oai/openaireopendoar:71602024-03-20T03:21:07.292504Repositório 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 Using cilia mutants to study left-right asymmetry in zebrafish
title Using cilia mutants to study left-right asymmetry in zebrafish
spellingShingle Using cilia mutants to study left-right asymmetry in zebrafish
Sampaio, Pedro Rafael Martins de Almeida
Kupffer’s vesicle
Motile cilia
KV flow
Left-right asymmetry
Dnah7
DeltaD mutant
title_short Using cilia mutants to study left-right asymmetry in zebrafish
title_full Using cilia mutants to study left-right asymmetry in zebrafish
title_fullStr Using cilia mutants to study left-right asymmetry in zebrafish
title_full_unstemmed Using cilia mutants to study left-right asymmetry in zebrafish
title_sort Using cilia mutants to study left-right asymmetry in zebrafish
author Sampaio, Pedro Rafael Martins de Almeida
author_facet Sampaio, Pedro Rafael Martins de Almeida
author_role author
dc.contributor.none.fl_str_mv Lopes, Susana
Ludovice, Ana
RUN
dc.contributor.author.fl_str_mv Sampaio, Pedro Rafael Martins de Almeida
dc.subject.por.fl_str_mv Kupffer’s vesicle
Motile cilia
KV flow
Left-right asymmetry
Dnah7
DeltaD mutant
topic Kupffer’s vesicle
Motile cilia
KV flow
Left-right asymmetry
Dnah7
DeltaD mutant
description A thesis submitted in fulfillment of the requirements for the degree of the Masters in Molecular Genetics and Biomedicine
publishDate 2014
dc.date.none.fl_str_mv 2014-09-30T10:37:31Z
2014
2014-01-01T00:00:00Z
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://hdl.handle.net/10362/13155
url http://hdl.handle.net/10362/13155
dc.language.iso.fl_str_mv eng
language eng
dc.rights.driver.fl_str_mv info:eu-repo/semantics/openAccess
eu_rights_str_mv openAccess
dc.format.none.fl_str_mv application/pdf
dc.publisher.none.fl_str_mv Faculdade de Ciências e Tecnologia
publisher.none.fl_str_mv Faculdade de Ciências e Tecnologia
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
instacron_str RCAAP
institution RCAAP
reponame_str Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)
collection Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)
repository.name.fl_str_mv Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) - Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação
repository.mail.fl_str_mv
_version_ 1799137852047491072

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