Nr2f1a maintains atrial nkx2.5 expression to repress pacemaker identity within venous atrial cardiomyocytes of zebrafish
Maintenance of cardiomyocyte identity is vital for normal heart development and function. However, our understanding of cardiomyocyte plasticity remains incomplete. Here, we show that sustained expression of the zebrafish transcription factor Nr2f1a prevents the progressive acquisition of ventricula...
| Main Authors: | , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
eLife Sciences Publications Ltd
2023-05-01
|
| Series: | eLife |
| Subjects: | |
| Online Access: | https://elifesciences.org/articles/77408 |
| _version_ | 1797826662042173440 |
|---|---|
| author | Kendall E Martin Padmapriyadarshini Ravisankar Manu Beerens Calum A MacRae Joshua S Waxman |
| author_facet | Kendall E Martin Padmapriyadarshini Ravisankar Manu Beerens Calum A MacRae Joshua S Waxman |
| author_sort | Kendall E Martin |
| collection | DOAJ |
| description | Maintenance of cardiomyocyte identity is vital for normal heart development and function. However, our understanding of cardiomyocyte plasticity remains incomplete. Here, we show that sustained expression of the zebrafish transcription factor Nr2f1a prevents the progressive acquisition of ventricular cardiomyocyte (VC) and pacemaker cardiomyocyte (PC) identities within distinct regions of the atrium. Transcriptomic analysis of flow-sorted atrial cardiomyocytes (ACs) from nr2f1a mutant zebrafish embryos showed increased VC marker gene expression and altered expression of core PC regulatory genes, including decreased expression of nkx2.5, a critical repressor of PC differentiation. At the arterial (outflow) pole of the atrium in nr2f1a mutants, cardiomyocytes resolve to VC identity within the expanded atrioventricular canal. However, at the venous (inflow) pole of the atrium, there is a progressive wave of AC transdifferentiation into PCs across the atrium toward the arterial pole. Restoring Nkx2.5 is sufficient to repress PC marker identity in nr2f1a mutant atria and analysis of chromatin accessibility identified an Nr2f1a-dependent nkx2.5 enhancer expressed in the atrial myocardium directly adjacent to PCs. CRISPR/Cas9-mediated deletion of the putative nkx2.5 enhancer leads to a loss of Nkx2.5-expressing ACs and expansion of a PC reporter, supporting that Nr2f1a limits PC differentiation within venous ACs via maintaining nkx2.5 expression. The Nr2f-dependent maintenance of AC identity within discrete atrial compartments may provide insights into the molecular etiology of concurrent structural congenital heart defects and associated arrhythmias. |
| first_indexed | 2024-04-09T12:35:59Z |
| format | Article |
| id | doaj.art-f836804f17944076be9c284e9dd8ddfc |
| institution | Directory Open Access Journal |
| issn | 2050-084X |
| language | English |
| last_indexed | 2024-04-09T12:35:59Z |
| publishDate | 2023-05-01 |
| publisher | eLife Sciences Publications Ltd |
| record_format | Article |
| series | eLife |
| spelling | doaj.art-f836804f17944076be9c284e9dd8ddfc2023-05-15T13:31:03ZengeLife Sciences Publications LtdeLife2050-084X2023-05-011210.7554/eLife.77408Nr2f1a maintains atrial nkx2.5 expression to repress pacemaker identity within venous atrial cardiomyocytes of zebrafishKendall E Martin0Padmapriyadarshini Ravisankar1Manu Beerens2Calum A MacRae3Joshua S Waxman4https://orcid.org/0000-0002-8132-487XMolecular Genetics, Biochemistry, and Microbiology Graduate Program, University of Cincinnati College of Medicine, Cincinnati, United States; Molecular Cardiovascular Biology Division and Heart Institute, Cincinnati Children’s Hospital Medical Center, Cincinnati, United StatesMolecular Cardiovascular Biology Division and Heart Institute, Cincinnati Children’s Hospital Medical Center, Cincinnati, United StatesDivisions of Cardiovascular Medicine, Genetics and Network Medicine, Department of Medicine, Brigham and Women's Hospital and Harvard Medical School, Boston, United StatesDivisions of Cardiovascular Medicine, Genetics and Network Medicine, Department of Medicine, Brigham and Women's Hospital and Harvard Medical School, Boston, United StatesMolecular Cardiovascular Biology Division and Heart Institute, Cincinnati Children’s Hospital Medical Center, Cincinnati, United States; Department of Pediatrics, University of Cincinnati College of Medicine, Cincinnati, United StatesMaintenance of cardiomyocyte identity is vital for normal heart development and function. However, our understanding of cardiomyocyte plasticity remains incomplete. Here, we show that sustained expression of the zebrafish transcription factor Nr2f1a prevents the progressive acquisition of ventricular cardiomyocyte (VC) and pacemaker cardiomyocyte (PC) identities within distinct regions of the atrium. Transcriptomic analysis of flow-sorted atrial cardiomyocytes (ACs) from nr2f1a mutant zebrafish embryos showed increased VC marker gene expression and altered expression of core PC regulatory genes, including decreased expression of nkx2.5, a critical repressor of PC differentiation. At the arterial (outflow) pole of the atrium in nr2f1a mutants, cardiomyocytes resolve to VC identity within the expanded atrioventricular canal. However, at the venous (inflow) pole of the atrium, there is a progressive wave of AC transdifferentiation into PCs across the atrium toward the arterial pole. Restoring Nkx2.5 is sufficient to repress PC marker identity in nr2f1a mutant atria and analysis of chromatin accessibility identified an Nr2f1a-dependent nkx2.5 enhancer expressed in the atrial myocardium directly adjacent to PCs. CRISPR/Cas9-mediated deletion of the putative nkx2.5 enhancer leads to a loss of Nkx2.5-expressing ACs and expansion of a PC reporter, supporting that Nr2f1a limits PC differentiation within venous ACs via maintaining nkx2.5 expression. The Nr2f-dependent maintenance of AC identity within discrete atrial compartments may provide insights into the molecular etiology of concurrent structural congenital heart defects and associated arrhythmias.https://elifesciences.org/articles/77408heart developmentatrial cardiomyocytescellular plasticityNr2f transcription factorpacemaker cardiomyocyteNkx2.5 transcription factor |
| spellingShingle | Kendall E Martin Padmapriyadarshini Ravisankar Manu Beerens Calum A MacRae Joshua S Waxman Nr2f1a maintains atrial nkx2.5 expression to repress pacemaker identity within venous atrial cardiomyocytes of zebrafish eLife heart development atrial cardiomyocytes cellular plasticity Nr2f transcription factor pacemaker cardiomyocyte Nkx2.5 transcription factor |
| title | Nr2f1a maintains atrial nkx2.5 expression to repress pacemaker identity within venous atrial cardiomyocytes of zebrafish |
| title_full | Nr2f1a maintains atrial nkx2.5 expression to repress pacemaker identity within venous atrial cardiomyocytes of zebrafish |
| title_fullStr | Nr2f1a maintains atrial nkx2.5 expression to repress pacemaker identity within venous atrial cardiomyocytes of zebrafish |
| title_full_unstemmed | Nr2f1a maintains atrial nkx2.5 expression to repress pacemaker identity within venous atrial cardiomyocytes of zebrafish |
| title_short | Nr2f1a maintains atrial nkx2.5 expression to repress pacemaker identity within venous atrial cardiomyocytes of zebrafish |
| title_sort | nr2f1a maintains atrial nkx2 5 expression to repress pacemaker identity within venous atrial cardiomyocytes of zebrafish |
| topic | heart development atrial cardiomyocytes cellular plasticity Nr2f transcription factor pacemaker cardiomyocyte Nkx2.5 transcription factor |
| url | https://elifesciences.org/articles/77408 |
| work_keys_str_mv | AT kendallemartin nr2f1amaintainsatrialnkx25expressiontorepresspacemakeridentitywithinvenousatrialcardiomyocytesofzebrafish AT padmapriyadarshiniravisankar nr2f1amaintainsatrialnkx25expressiontorepresspacemakeridentitywithinvenousatrialcardiomyocytesofzebrafish AT manubeerens nr2f1amaintainsatrialnkx25expressiontorepresspacemakeridentitywithinvenousatrialcardiomyocytesofzebrafish AT calumamacrae nr2f1amaintainsatrialnkx25expressiontorepresspacemakeridentitywithinvenousatrialcardiomyocytesofzebrafish AT joshuaswaxman nr2f1amaintainsatrialnkx25expressiontorepresspacemakeridentitywithinvenousatrialcardiomyocytesofzebrafish |