Live imaging of adult zebrafish cardiomyocyte proliferation ex vivo

Hessel Honkoop, Phong D Nguyen, Veronique E M van der Velden, Katharina F Sonnen, Jeroen Bakkers

Research output: Contribution to journal/periodicalArticleScientificpeer-review

Abstract

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.

Original languageEnglish
JournalDevelopment (Cambridge)
Volume148
Issue number18
DOIs
Publication statusPublished - 15 Sep 2021

Keywords

  • Animals
  • Animals, Genetically Modified/metabolism
  • Calpain/metabolism
  • Cell Nucleus/metabolism
  • Cell Proliferation/physiology
  • Cells, Cultured
  • Cytokinesis/physiology
  • Female
  • Heart/physiology
  • Male
  • Mammals/metabolism
  • Myocytes, Cardiac/metabolism
  • Proteasome Endopeptidase Complex/metabolism
  • Regeneration/physiology
  • Sarcomeres/metabolism
  • Zebrafish/metabolism
  • Zebrafish Proteins/metabolism

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