Optimized protocol for live-cell analysis of kinetochore fiber maturation in Indian muntjac cells
Summary: Here, we take advantage of the low chromosome number (2N=6) and distinctively large kinetochores of female Indian muntjac cells to investigate the molecular mechanism underlying k-fiber maturation. We describe steps for monitoring kinetochore-microtubule dynamics over time. Specifically, we...
| Main Authors: | , , |
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| Format: | Article |
| Language: | English |
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Elsevier
2023-03-01
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| Series: | STAR Protocols |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S2666166722008917 |
| _version_ | 1811177258325377024 |
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| author | Ana Coelho Almeida Joana Soares-de-Oliveira Helder Maiato |
| author_facet | Ana Coelho Almeida Joana Soares-de-Oliveira Helder Maiato |
| author_sort | Ana Coelho Almeida |
| collection | DOAJ |
| description | Summary: Here, we take advantage of the low chromosome number (2N=6) and distinctively large kinetochores of female Indian muntjac cells to investigate the molecular mechanism underlying k-fiber maturation. We describe steps for monitoring kinetochore-microtubule dynamics over time. Specifically, we detail the combination of live-cell super-resolution CH-STED microscopy of microtubule growth events within individual k-fibers and a laser-mediated k-fiber injury/repair assay. These tools provide a direct assessment of microtubule amplification mechanisms within k-fibers in metazoans.For complete details on the use and execution of this protocol, please refer to Almeida et al. (2022).1 : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics. |
| first_indexed | 2024-04-10T22:58:42Z |
| format | Article |
| id | doaj.art-14877f1d407447fdafdeb1e922d89275 |
| institution | Directory Open Access Journal |
| issn | 2666-1667 |
| language | English |
| last_indexed | 2024-04-10T22:58:42Z |
| publishDate | 2023-03-01 |
| publisher | Elsevier |
| record_format | Article |
| series | STAR Protocols |
| spelling | doaj.art-14877f1d407447fdafdeb1e922d892752023-01-14T04:27:11ZengElsevierSTAR Protocols2666-16672023-03-0141102011Optimized protocol for live-cell analysis of kinetochore fiber maturation in Indian muntjac cellsAna Coelho Almeida0Joana Soares-de-Oliveira1Helder Maiato2Instituto de Investigação e Inovação em Saúde (i3S), Universidade do Porto, Rua Alfredo Allen 208, 4200-135 Porto, Portugal; Instituto de Biologia Molecular e Celular, Universidade do Porto, Rua Alfredo Allen 208, 4200-135 Porto, PortugalInstituto de Investigação e Inovação em Saúde (i3S), Universidade do Porto, Rua Alfredo Allen 208, 4200-135 Porto, Portugal; Instituto de Biologia Molecular e Celular, Universidade do Porto, Rua Alfredo Allen 208, 4200-135 Porto, PortugalInstituto de Investigação e Inovação em Saúde (i3S), Universidade do Porto, Rua Alfredo Allen 208, 4200-135 Porto, Portugal; Instituto de Biologia Molecular e Celular, Universidade do Porto, Rua Alfredo Allen 208, 4200-135 Porto, Portugal; Cell Division Group, Department of Biomedicine, Faculdade de Medicina, Universidade do Porto, Alameda Professor Hernâni Monteiro, 4200-319 Porto, Portugal; Corresponding authorSummary: Here, we take advantage of the low chromosome number (2N=6) and distinctively large kinetochores of female Indian muntjac cells to investigate the molecular mechanism underlying k-fiber maturation. We describe steps for monitoring kinetochore-microtubule dynamics over time. Specifically, we detail the combination of live-cell super-resolution CH-STED microscopy of microtubule growth events within individual k-fibers and a laser-mediated k-fiber injury/repair assay. These tools provide a direct assessment of microtubule amplification mechanisms within k-fibers in metazoans.For complete details on the use and execution of this protocol, please refer to Almeida et al. (2022).1 : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.http://www.sciencedirect.com/science/article/pii/S2666166722008917Cell BiologyMicroscopy |
| spellingShingle | Ana Coelho Almeida Joana Soares-de-Oliveira Helder Maiato Optimized protocol for live-cell analysis of kinetochore fiber maturation in Indian muntjac cells STAR Protocols Cell Biology Microscopy |
| title | Optimized protocol for live-cell analysis of kinetochore fiber maturation in Indian muntjac cells |
| title_full | Optimized protocol for live-cell analysis of kinetochore fiber maturation in Indian muntjac cells |
| title_fullStr | Optimized protocol for live-cell analysis of kinetochore fiber maturation in Indian muntjac cells |
| title_full_unstemmed | Optimized protocol for live-cell analysis of kinetochore fiber maturation in Indian muntjac cells |
| title_short | Optimized protocol for live-cell analysis of kinetochore fiber maturation in Indian muntjac cells |
| title_sort | optimized protocol for live cell analysis of kinetochore fiber maturation in indian muntjac cells |
| topic | Cell Biology Microscopy |
| url | http://www.sciencedirect.com/science/article/pii/S2666166722008917 |
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