Imaging the early zebrafish embryo centrosomes following injection of small-molecule inhibitors to understand spindle formation
| العنوان: | Imaging the early zebrafish embryo centrosomes following injection of small-molecule inhibitors to understand spindle formation |
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| المؤلفون: | Thomas Cammerino, Heidi Hehnly, Lindsay Rathbun, Abrar A. Aljiboury, Amra Mujcic |
| المصدر: | STAR Protocols STAR Protocols, Vol 2, Iss 1, Pp 100293-(2021) |
| سنة النشر: | 2021 |
| مصطلحات موضوعية: | Model organisms, Cell biology, food.ingredient, animal structures, Embryo, Nonmammalian, ved/biology.organism_classification_rank.species, Mitosis, Spindle Apparatus, Biology, General Biochemistry, Genetics and Molecular Biology, law.invention, food, Confocal microscopy, law, Yolk, Protocol, Animals, Model organism, lcsh:Science (General), Protein Kinase Inhibitors, Zebrafish, Centrosome, Microscopy, Microscopy, Confocal, General Immunology and Microbiology, ved/biology, General Neuroscience, Embryo, Embryonic stem cell, Spindle apparatus, embryonic structures, Multipolar spindles, lcsh:Q1-390 |
| الوصف: | Summary During the earliest division stages, zebrafish embryos have large cells that divide rapidly and synchronously to create a cellular layer on top of the yolk. Here, we describe a protocol for monitoring spindle dynamics during these early embryonic divisions. We outline techniques for injecting zebrafish embryos with small-molecule inhibitors toward polo-like kinases, preparing and mounting embryos for three-dimensional imaging using confocal microscopy. These techniques are used to understand how the early zebrafish embryo’s centrosome constructs the mitotic spindle. For complete details on the use and execution of this protocol, please refer to Rathbun et al. (2020). Graphical abstract Highlights • Breeding and obtaining zebrafish embryos to monitor spindle dynamics • Zebrafish embryo microinjection to introduce polo-like kinase (PLK1/4) inhibitors • Mounting of live and fixed zebrafish embryo for early embryonic studies • Imaging live and fixed zebrafish embryos using confocal microscope During the earliest division stages, zebrafish embryos have large cells that divide rapidly and synchronously to create a cellular layer on top of the yolk. Here, we describe a protocol for monitoring spindle dynamics during these early embryonic divisions. We outline techniques for injecting zebrafish embryos with small-molecule inhibitors toward polo-like kinases, preparing and mounting embryos for three-dimensional imaging using confocal microscopy. These techniques are used to understand how the early zebrafish embryo’s centrosome constructs the mitotic spindle. |
| تدمد: | 2666-1667 |
| URL الوصول: | https://explore.openaire.eu/search/publication?articleId=doi_dedup___::130f99423aa1d61d11844df9503cabd2 https://pubmed.ncbi.nlm.nih.gov/33554134 |
| حقوق: | OPEN |
| رقم الأكسشن: | edsair.doi.dedup.....130f99423aa1d61d11844df9503cabd2 |
| قاعدة البيانات: | OpenAIRE |
| تدمد: | 26661667 |
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