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
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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