Analysis of cardiomyocyte development using immunofluorescence in embryonic mouse heart

Lisa D. Wilsbacher*, Shaun R. Coughlin

*Corresponding author for this work

Research output: Contribution to journalArticle

5 Citations (Scopus)

Abstract

During heart development, the generation of myocardial-specific structural and functional units including sarcomeres, contractile myofibrils, intercalated discs, and costameres requires the coordinated assembly of multiple components in time and space. Disruption in assembly of these components leads to developmental heart defects. Immunofluorescent staining techniques are used commonly in cultured cardiomyocytes to probe myofibril maturation, but this ex vivo approach is limited by the extent to which myocytes will fully differentiate in culture, lack of normal in vivo mechanical inputs, and absence of endocardial cues. Application of immunofluorescence techniques to the study of developing mouse heart is desirable but more technically challenging, and methods often lack sufficient sensitivity and resolution to visualize sarcomeres in the early stages of heart development. Here, we describe a robust and reproducible method to co-immunostain multiple proteins or to co-visualize a fluorescent protein with immunofluorescent staining in the embryonic mouse heart and use this method to analyze developing myofibrils, intercalated discs, and costameres. This method can be further applied to assess cardiomyocyte structural changes caused by mutations that lead to developmental heart defects.

Original languageEnglish (US)
JournalJournal of Visualized Experiments
Issue number97
DOIs
StatePublished - Mar 26 2015

Fingerprint

Cardiac Myocytes
Fluorescent Antibody Technique
Myofibrils
Costameres
Sarcomeres
Staining and Labeling
Proteins
Defects
Muscle Cells
Cues
Mutation

Keywords

  • Cardiomyocyte
  • Costamere
  • Cryosection
  • Development
  • Developmental Biology
  • Embryonic heart
  • Immunofluorescence
  • Intercalated disc
  • Issue 97
  • Mouse
  • S-α-actinin
  • Sarcomere

ASJC Scopus subject areas

  • Neuroscience(all)
  • Chemical Engineering(all)
  • Immunology and Microbiology(all)
  • Biochemistry, Genetics and Molecular Biology(all)

Cite this

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Analysis of cardiomyocyte development using immunofluorescence in embryonic mouse heart. / Wilsbacher, Lisa D.; Coughlin, Shaun R.

In: Journal of Visualized Experiments, No. 97, 26.03.2015.

Research output: Contribution to journalArticle

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