Gon4l/Udu regulates cardiomyocyte proliferation and maintenance of ventricular chamber identity during zebrafish development

• Published in: Developmental biology Vol. 462; no. 2; pp. 223 - 234
• Main Authors: Budine, Terin E., de Sena-Tomás, Carmen, Williams, Margot L.K., Sepich, Diane S., Targoff, Kimara L., Solnica-Krezel, Lila
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 15.06.2020
• Subjects: Animals; Cell Differentiation - physiology; Cell Proliferation - physiology; Chamber identity; Chromatin - metabolism; Erythroid-Specific DNA-Binding Factors - metabolism; Erythroid-Specific DNA-Binding Factors - physiology; Gene Expression Regulation, Developmental - genetics; Heart - embryology; Heart Atria - embryology; Heart Atria - metabolism; Heart development; Myocardium - metabolism; Myocytes, Cardiac - metabolism; Myocytes, Cardiac - physiology; nkx2.5; Proliferation; S Phase - genetics; Transcription Factors - metabolism; Ugly duckling (udu); Zebrafish - embryology; Zebrafish - genetics; Zebrafish - metabolism; Zebrafish Proteins - metabolism; Zebrafish Proteins - physiology; Chamber identity; Ugly duckling (udu); Proliferation; Heart development; nkx2.5
• ISSN: 0012-1606; 1095-564X
• DOI: 10.1016/j.ydbio.2020.03.002

Vertebrate heart development requires spatiotemporal regulation of gene expression to specify cardiomyocytes, increase the cardiomyocyte population through proliferation, and to establish and maintain atrial and ventricular cardiac chamber identities. The evolutionarily conserved chromatin factor Gon4-like (Gon4l), encoded by the zebrafish ugly duckling (udu) locus, has previously been implicated in cell proliferation, cell survival, and specification of mesoderm-derived tissues including blood and somites, but its role in heart formation has not been studied. Here we report two distinct roles of Gon4l/Udu in heart development: regulation of cell proliferation and maintenance of ventricular identity. We show that zygotic loss of udu expression causes a significant reduction in cardiomyocyte number at one day post fertilization that becomes exacerbated during later development. We present evidence that the cardiomyocyte deficiency in udu mutants results from reduced cell proliferation, unlike hematopoietic deficiencies attributed to TP53-dependent apoptosis. We also demonstrate that expression of the G1/S-phase cell cycle regulator, cyclin E2 (ccne2), is reduced in udu mutant hearts, and that the Gon4l protein associates with regulatory regions of the ccne2 gene during early embryogenesis. Furthermore, udu mutant hearts exhibit a decrease in the proportion of ventricular cardiomyocytes compared to atrial cardiomyocytes, concomitant with progressive reduction of nkx2.5 expression. We further demonstrate that udu and nkx2.5 interact to maintain the proportion of ventricular cardiomyocytes during development. However, we find that ectopic expression of nkx2.5 is not sufficient to restore ventricular chamber identity suggesting that Gon4l regulates cardiac chamber patterning via multiple pathways. Together, our findings define a novel role for zygotically-expressed Gon4l in coordinating cardiomyocyte proliferation and chamber identity maintenance during cardiac development. •Gon4l functions in cardiomyocyte proliferation and ventricular fate maintenance.•Reduced number of cardiomyocytes in udu−/− embryos is TP53-apoptosis independent.•Cardiomyocytes in udu−/− embryos fail to proliferation after heart tube formation.•Expression of known cell cycle regulator cnne1 is reduced in udu−/− cardiomyocytes.•Udu and nkx2.5 genetically in ventricular chamber identity maintenance.