Protocols for Investigating the Epithelial Properties of Cardiac Progenitor Cells in the Mouse Embryo

Claudio Cortes, Christopher De Bono, Charlotte Thellier, Alexandre Francou, Robert G. Kelly

Research output: Chapter in Book/Report/Conference proceedingChapter

Abstract

Epithelial cardiac progenitor cells of the second heart field (SHF) contribute to growth of the vertebrate heart tube by progressive addition of cells from the dorsal pericardial wall to the cardiac poles. Perturbation of SHF development, including defects in apicobasal or planar polarity, results in shortening of the heart tube and a spectrum of congenital heart defects. Here, we provide detailed protocols for fixed section and wholemount immunofluorescence and live imaging approaches to studying the epithelial properties of cardiac progenitors in the dorsal pericardial wall during mouse heart development. Whole-embryo culture and electroporation methods are also presented, allowing for pharmacological and genetic perturbation of SHF development, as well as image analysis approaches to quantify cell features across the progenitor cell epithelium. These protocols are broadly applicable to the study of epithelia in the early embryo.

Original languageEnglish (US)
Title of host publicationMethods in Molecular Biology
PublisherHumana Press Inc.
Pages231-250
Number of pages20
DOIs
StatePublished - 2022

Publication series

NameMethods in Molecular Biology
Volume2438
ISSN (Print)1064-3745
ISSN (Electronic)1940-6029

Keywords

  • Electroporation
  • Embryo culture
  • Epithelium
  • Heart development
  • Immunofluorescence
  • Second heart field
  • Time-lapse imaging

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

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