Ex Vivo Techniques to Study Heart Regeneration in Zebrafish

Methods Mol Biol. 2021;2158:211-222. doi: 10.1007/978-1-0716-0668-1_16.

Abstract

Due to its pronounced regenerative capacity, the zebrafish heart represents an advantageous model system for exploring the cellular and molecular mechanisms of cardiac regeneration. Upon injury, the epicardium, the outermost mesothelial tissue layer of vertebrate hearts, serves dual purposes in the regenerating heart as both a signaling center and a source for crucial cell types. Traditional in vivo genetic approaches to study heart regeneration can be time consuming and are not applicable to large-scale approaches and live surveillance of cellular behaviors. Here, we demonstrate ex vivo methods to culture, maintain, and study the regenerative responses of epicardial tissue in excised zebrafish hearts. Epicardial cell proliferation and migration are monitored in real time after uninjured or injured hearts are excised, washed, and cultured for up to 30 days. In addition to these techniques, we describe ex vivo genetic ablation of the epicardium, cell proliferation assays, partial ventricular explant culturing, and chemical screening.

Keywords: Chemical screening; Epicardium; Heart explant culture; Heart regeneration; Live imaging; Zebrafish.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Animals, Genetically Modified
  • Anti-Infective Agents / pharmacology
  • Cell Movement
  • Cell Proliferation
  • Heart / drug effects
  • Heart / growth & development*
  • High-Throughput Screening Assays / methods*
  • Metronidazole / pharmacology*
  • Myocytes, Cardiac / cytology*
  • Myocytes, Cardiac / drug effects
  • Pericardium / cytology*
  • Pericardium / drug effects
  • Regeneration*
  • Zebrafish
  • Zebrafish Proteins / genetics*

Substances

  • Anti-Infective Agents
  • Zebrafish Proteins
  • Metronidazole