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Cardiomyocyte differentiation by GATA-4-deficient embryonic stem cells
N Narita1, M Bielinska, D B Wilson
1Department of Pediatrics, Washington University School of Medicine, St. Louis Children's Hospital, MO 63110, USA.
Development (Cambridge, England)
|November 21, 1997
Summary
Transcription factor GATA-4 is not essential for cardiomyocyte differentiation. While Gata4-/- embryonic stem cells formed cardiomyocytes in vitro and in vivo, GATA-4 absence suggests other GATA-binding proteins may compensate.
Area of Science:
- Cardiovascular Biology
- Developmental Biology
- Stem Cell Biology
Background:
- Transcription factor GATA-4 has been implicated in cardiomyocyte differentiation.
- Previous studies suggested GATA-4's essential role in cardiac lineage commitment.
Purpose of the Study:
- To investigate whether GATA-4 is essential for cardiomyocyte lineage commitment using Gata4-/- embryonic stem (ES) cells.
- To assess the in vitro and in vivo differentiation capacity of Gata4-/- ES cells into cardiomyocytes.
Main Methods:
- In vitro differentiation of embryoid bodies from wild-type and Gata4-/- ES cells.
- In vivo analysis using chimeric mice generated by injecting Gata4-/- ES cells into 8-cell-stage embryos.
- Assessment of cardiomyocyte formation via microscopy, immunofluorescence, and in situ hybridization.
Main Results:
- Gata4-/- ES cells formed contracting cardiomyocytes in vitro, although at a lower frequency than wild-type.
- Electron microscopy and immunofluorescence confirmed sarcomere and cardiac myosin presence in mutant cardiomyocytes.
- Chimeric mice analysis showed Gata4-/- ES cells contributed to endocardium, myocardium, and epicardium, expressing cardiac-specific genes.
Conclusions:
- GATA-4 is not essential for terminal cardiomyocyte differentiation.
- Other GATA-binding proteins, such as GATA-5 or GATA-6, may compensate for the absence of GATA-4 in cardiac development.