Live imaging of adult zebrafish cardiomyocyte proliferation ex vivo
Autor: | Katharina F Sonnen, Phong D. Nguyen, Jeroen Bakkers, Veronique E M van der Velden, Hessel Honkoop |
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Přispěvatelé: | Hubrecht Institute for Developmental Biology and Stem Cell Research |
Rok vydání: | 2021 |
Předmět: |
Male
Research Report Zebrafish Proteins/metabolism Proliferation Sarcomere Animals Genetically Modified Sarcomeres/metabolism Myocytes Cardiac Cardiomyocyte proliferation Zebrafish Cells Cultured Mammals Cultured biology Mammals/metabolism Calpain Calpain/metabolism Cytokinesis/physiology Heart Myocytes Cardiac/metabolism Cell biology Heart/physiology Cardiac/metabolism Female Cardiac Regeneration/physiology Sarcomeres Proteasome Endopeptidase Complex Cardiovascular Development and Regeneration Cells Cell Proliferation/physiology Live cell imaging Zebrafish/metabolism Animals Regeneration Molecular Biology Genetically Modified/metabolism Cell Proliferation Cytokinesis Cell Nucleus Myocytes Live-imaging Regeneration (biology) Animals Genetically Modified/metabolism Zebrafish Proteins biology.organism_classification Proteasome Endopeptidase Complex/metabolism Cell Nucleus/metabolism biology.protein Ex vivo Developmental Biology |
Zdroj: | Development (Cambridge, England) article-version (VoR) Version of Record Development (Cambridge), 148(18). Company of Biologists Ltd |
ISSN: | 1477-9129 0950-1991 |
Popis: | Zebrafish are excellent at regenerating their heart by reinitiating proliferation in pre-existing cardiomyocytes. Studying how zebrafish achieve this holds great potential in developing new strategies to boost mammalian heart regeneration. Nevertheless, the lack of appropriate live-imaging tools for the adult zebrafish heart has limited detailed studies into the dynamics underlying cardiomyocyte proliferation. Here, we address this by developing a system in which cardiac slices of the injured zebrafish heart are cultured ex vivo for several days while retaining key regenerative characteristics, including cardiomyocyte proliferation. In addition, we show that the cardiac slice culture system is compatible with live timelapse imaging and allows manipulation of regenerating cardiomyocytes with drugs that normally would have toxic effects that prevent their use. Finally, we use the cardiac slices to demonstrate that adult cardiomyocytes with fully assembled sarcomeres can partially disassemble their sarcomeres in a calpain- and proteasome-dependent manner to progress through nuclear division and cytokinesis. In conclusion, we have developed a cardiac slice culture system, which allows imaging of native cardiomyocyte dynamics in real time to discover cellular mechanisms during heart regeneration. Summary: A method that allows live imaging of proliferating cardiomyocytes during zebrafish heart regeneration in their native tissue context. |
Databáze: | OpenAIRE |
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