Long-term, in toto live imaging of cardiomyocyte behaviour during mouse ventricle chamber formation at single-cell resolution

Nat Cell Biol. 2020 Mar;22(3):332-340. doi: 10.1038/s41556-020-0475-2. Epub 2020 Mar 2.

Abstract

Mapping of the holistic cell behaviours sculpting the four-chambered mammalian heart has been a goal or previous studies, but so far only success in transparent invertebrates and lower vertebrates with two-chambered hearts has been achieved. Using a live-imaging system comprising a customized vertical light-sheet microscope equipped with a mouse embryo culture module, a heartbeat-gated imaging strategy and a digital image processing framework, we realized volumetric imaging of developing mouse hearts at single-cell resolution and with uninterrupted cell lineages for up to 1.5 d. Four-dimensional landscapes of Nppa+ cardiomyocyte cell behaviours revealed a blueprint for ventricle chamber formation by which biased outward migration of the outermost cardiomyocytes is coupled with cell intercalation and horizontal division. The inner-muscle architecture of trabeculae was developed through dual mechanisms: early fate segregation and transmural cell arrangement involving both oriented cell division and directional migration. Thus, live-imaging reconstruction of uninterrupted cell lineages affords a transformative means for deciphering mammalian organogenesis.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Cell Division
  • Cell Lineage
  • Cell Movement
  • Embryo, Mammalian / cytology
  • Embryonic Development
  • Heart / embryology
  • Heart Ventricles / cytology*
  • Heart Ventricles / embryology*
  • Image Processing, Computer-Assisted / methods*
  • Mice
  • Microscopy
  • Morphogenesis
  • Myocardium / cytology
  • Myocytes, Cardiac / cytology*
  • Single-Cell Analysis
  • Tissue Culture Techniques