Drosophila S2 cells as a model system to investigate mitotic spindle dynamics, architecture, and function
Autor(a) principal: | |
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Data de Publicação: | 2010 |
Outros Autores: | , |
Tipo de documento: | Artigo |
Idioma: | eng |
Título da fonte: | Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) |
Texto Completo: | http://hdl.handle.net/10216/53787 |
Resumo: | In order to perpetuate their genetic content, eukaryotic cells have developed a microtubule-based machine known as the mitotic spindle. Independently of the system studied, mitotic spindles share at least one common characteristic – the dynamic nature of microtubules. This property allows the constant plasticity needed to assemble a bipolar structure, make proper kinetochoremicrotubule attachments, segregate chromosomes and finally disassemble the spindle and reform an interphase microtubule array. Here we describe a variety of experimental approaches currently used in our laboratory to study microtubule dynamics during mitosis using Drosophila melanogaster S2 cells as a model. By using quantitative live-cell imaging microscopy in combination with an advantageous labeling background, we illustrate how several cooperative pathways are used to build functional mitotic spindles. We illustrate different ways of perturbing spindle microtubule dynamics, including pharmacological inhibition and RNA interference of proteins that directly or indirectly impair microtubule dynamics. Additionally, we demonstrate the advantage of using fluorescent speckle microscopy (FSM) to investigate an intrinsic property of spindle microtubules known as poleward flux. Finally, we developed a set of laser microsurgery-based experiments that allow, with unique spatiotemporal resolution, the study of specific spindle-structures (e.g. centrosomes, microtubules and kinetochores) and their respective roles during mitosis. |
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Drosophila S2 cells as a model system to investigate mitotic spindle dynamics, architecture, and functionLive-cell microscopyLaser microsurgeryFluorescent speckle microscopyMicrotubulesMicrotubule drugsIn order to perpetuate their genetic content, eukaryotic cells have developed a microtubule-based machine known as the mitotic spindle. Independently of the system studied, mitotic spindles share at least one common characteristic – the dynamic nature of microtubules. This property allows the constant plasticity needed to assemble a bipolar structure, make proper kinetochoremicrotubule attachments, segregate chromosomes and finally disassemble the spindle and reform an interphase microtubule array. Here we describe a variety of experimental approaches currently used in our laboratory to study microtubule dynamics during mitosis using Drosophila melanogaster S2 cells as a model. By using quantitative live-cell imaging microscopy in combination with an advantageous labeling background, we illustrate how several cooperative pathways are used to build functional mitotic spindles. We illustrate different ways of perturbing spindle microtubule dynamics, including pharmacological inhibition and RNA interference of proteins that directly or indirectly impair microtubule dynamics. Additionally, we demonstrate the advantage of using fluorescent speckle microscopy (FSM) to investigate an intrinsic property of spindle microtubules known as poleward flux. Finally, we developed a set of laser microsurgery-based experiments that allow, with unique spatiotemporal resolution, the study of specific spindle-structures (e.g. centrosomes, microtubules and kinetochores) and their respective roles during mitosis.20102010-01-01T00:00:00Zinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/articleapplication/pdfhttp://hdl.handle.net/10216/53787eng0091-679XMoutinho-Pereira, SMatos, IMaiato, Hinfo: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:RCAAP2023-11-29T13:35:27Zoai:repositorio-aberto.up.pt:10216/53787Portal AgregadorONGhttps://www.rcaap.pt/oai/openaireopendoar:71602024-03-19T23:43:18.565963Repositório 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 |
Drosophila S2 cells as a model system to investigate mitotic spindle dynamics, architecture, and function |
title |
Drosophila S2 cells as a model system to investigate mitotic spindle dynamics, architecture, and function |
spellingShingle |
Drosophila S2 cells as a model system to investigate mitotic spindle dynamics, architecture, and function Moutinho-Pereira, S Live-cell microscopy Laser microsurgery Fluorescent speckle microscopy Microtubules Microtubule drugs |
title_short |
Drosophila S2 cells as a model system to investigate mitotic spindle dynamics, architecture, and function |
title_full |
Drosophila S2 cells as a model system to investigate mitotic spindle dynamics, architecture, and function |
title_fullStr |
Drosophila S2 cells as a model system to investigate mitotic spindle dynamics, architecture, and function |
title_full_unstemmed |
Drosophila S2 cells as a model system to investigate mitotic spindle dynamics, architecture, and function |
title_sort |
Drosophila S2 cells as a model system to investigate mitotic spindle dynamics, architecture, and function |
author |
Moutinho-Pereira, S |
author_facet |
Moutinho-Pereira, S Matos, I Maiato, H |
author_role |
author |
author2 |
Matos, I Maiato, H |
author2_role |
author author |
dc.contributor.author.fl_str_mv |
Moutinho-Pereira, S Matos, I Maiato, H |
dc.subject.por.fl_str_mv |
Live-cell microscopy Laser microsurgery Fluorescent speckle microscopy Microtubules Microtubule drugs |
topic |
Live-cell microscopy Laser microsurgery Fluorescent speckle microscopy Microtubules Microtubule drugs |
description |
In order to perpetuate their genetic content, eukaryotic cells have developed a microtubule-based machine known as the mitotic spindle. Independently of the system studied, mitotic spindles share at least one common characteristic – the dynamic nature of microtubules. This property allows the constant plasticity needed to assemble a bipolar structure, make proper kinetochoremicrotubule attachments, segregate chromosomes and finally disassemble the spindle and reform an interphase microtubule array. Here we describe a variety of experimental approaches currently used in our laboratory to study microtubule dynamics during mitosis using Drosophila melanogaster S2 cells as a model. By using quantitative live-cell imaging microscopy in combination with an advantageous labeling background, we illustrate how several cooperative pathways are used to build functional mitotic spindles. We illustrate different ways of perturbing spindle microtubule dynamics, including pharmacological inhibition and RNA interference of proteins that directly or indirectly impair microtubule dynamics. Additionally, we demonstrate the advantage of using fluorescent speckle microscopy (FSM) to investigate an intrinsic property of spindle microtubules known as poleward flux. Finally, we developed a set of laser microsurgery-based experiments that allow, with unique spatiotemporal resolution, the study of specific spindle-structures (e.g. centrosomes, microtubules and kinetochores) and their respective roles during mitosis. |
publishDate |
2010 |
dc.date.none.fl_str_mv |
2010 2010-01-01T00:00:00Z |
dc.type.status.fl_str_mv |
info:eu-repo/semantics/publishedVersion |
dc.type.driver.fl_str_mv |
info:eu-repo/semantics/article |
format |
article |
status_str |
publishedVersion |
dc.identifier.uri.fl_str_mv |
http://hdl.handle.net/10216/53787 |
url |
http://hdl.handle.net/10216/53787 |
dc.language.iso.fl_str_mv |
eng |
language |
eng |
dc.relation.none.fl_str_mv |
0091-679X |
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.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 |
|
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1799135747749445632 |