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      Tbx20 Is an Essential Regulator of Embryonic Heart Growth in Zebrafish

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          Abstract

          The molecular mechanisms that regulate cardiomyocyte proliferation during embryonic heart growth are not completely deciphered yet. In a forward genetic N-ethyl-N-nitrosourea (ENU) mutagenesis screen, we identified the recessive embryonic-lethal zebrafish mutant line weiches herz ( whz). Homozygous mutant whz embryos display impaired heart growth due to diminished embryonic cardiomyocyte proliferation resulting in cardiac hypoplasia and weak cardiac contraction. By positional cloning, we found in whz mutant zebrafish a missense mutation within the T-box 20 (Tbx20) transcription factor gene leading to destabilization of Tbx20 protein. Morpholino-mediated knock-down of Tbx20 in wild-type zebrafish embryos phenocopies whz, indicating that the whz phenotype is due to loss of Tbx20 function, thereby leading to significantly reduced cardiomyocyte numbers by impaired proliferation of heart muscle cells. Ectopic overexpression of wild-type Tbx20 in whz mutant embryos restored cardiomyocyte proliferation and heart growth. Interestingly, ectopic overexpression of Tbx20 in wild-type zebrafish embryos resulted, similar to the situation in the embryonic mouse heart, in significantly reduced proliferation rates of ventricular cardiomyocytes, suggesting that Tbx20 activity needs to be tightly fine-tuned to guarantee regular cardiomyocyte proliferation and embryonic heart growth in vivo.

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          Most cited references28

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          In situ hybridization: an improved whole-mount method for Xenopus embryos.

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            Quality control of eukaryotic mRNA: safeguarding cells from abnormal mRNA function.

            Cells routinely make mistakes. Some mistakes are encoded by the genome and may manifest as inherited or acquired diseases. Other mistakes occur because metabolic processes can be intrinsically inefficient or inaccurate. Consequently, cells have developed mechanisms to minimize the damage that would result if mistakes went unchecked. Here, we provide an overview of three quality control mechanisms--nonsense-mediated mRNA decay, nonstop mRNA decay, and no-go mRNA decay. Each surveys mRNAs during translation and degrades those mRNAs that direct aberrant protein synthesis. Along with other types of quality control that occur during the complex processes of mRNA biogenesis, these mRNA surveillance mechanisms help to ensure the integrity of protein-encoding gene expression.
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              Building and re-building the heart by cardiomyocyte proliferation.

              The adult human heart does not regenerate significant amounts of lost tissue after injury. Rather than making new, functional muscle, human hearts are prone to scarring and hypertrophy, which can often lead to fatal arrhythmias and heart failure. The most-cited basis of this ineffective cardiac regeneration in mammals is the low proliferative capacity of adult cardiomyocytes. However, mammalian cardiomyocytes can avidly proliferate during fetal and neonatal development, and both adult zebrafish and neonatal mice can regenerate cardiac muscle after injury, suggesting that latent regenerative potential exists. Dissecting the cellular and molecular mechanisms that promote cardiomyocyte proliferation throughout life, deciphering why proliferative capacity normally dissipates in adult mammals, and deriving means to boost this capacity are primary goals in cardiovascular research. Here, we review our current understanding of how cardiomyocyte proliferation is regulated during heart development and regeneration.
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                Author and article information

                Contributors
                Role: Editor
                Journal
                PLoS One
                PLoS ONE
                plos
                plosone
                PLoS ONE
                Public Library of Science (San Francisco, CA USA )
                1932-6203
                1 December 2016
                2016
                : 11
                : 12
                : e0167306
                Affiliations
                [1 ]Molecular Cardiology, Department of Medicine II, University of Ulm, Ulm, Germany
                [2 ]Department of Medicine II, University of Ulm, Ulm, Germany
                Academia Sinica, TAIWAN
                Author notes

                Competing Interests: The authors have declared that no competing interests exist.

                • Conceptualization: SJ WR.

                • Funding acquisition: SJ WR.

                • Investigation: SJ LR IMB AB MK.

                • Methodology: SJ LR IMB.

                • Supervision: SJ WR.

                • Validation: SJ WR AB.

                • Visualization: SJ LR WR.

                • Writing – original draft: SJ LR WR.

                • Writing – review & editing: SJ WR AB MK.

                Article
                PONE-D-16-34221
                10.1371/journal.pone.0167306
                5132222
                27907103
                94a728b1-44b3-4476-bd66-d1aef6add484
                © 2016 Just et al

                This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

                History
                : 26 August 2016
                : 13 November 2016
                Page count
                Figures: 6, Tables: 0, Pages: 17
                Funding
                Funded by: funder-id http://dx.doi.org/10.13039/501100001659, Deutsche Forschungsgemeinschaft;
                Award ID: RO2173/3-1; RO2173/3-2
                Award Recipient :
                Funded by: funder-id http://dx.doi.org/10.13039/501100001659, Deutsche Forschungsgemeinschaft;
                Award ID: JU2859/2-1
                Award Recipient :
                Funded by: funder-id http://dx.doi.org/10.13039/501100002347, Bundesministerium für Bildung und Forschung;
                Award ID: 01ZX1407A
                Award Recipient :
                Funded by: Bundesministerium für Bildung und Forschung (DE)
                Award ID: 01GS0108; 01GS0420; 01GS0836; 01GS1104; 01KU0901C
                Award Recipient :
                Funded by: European Union
                Award ID: Inheritance
                Award Recipient :
                Funded by: funder-id http://dx.doi.org/10.13039/501100003542, Ministerium für Wissenschaft, Forschung und Kunst Baden-Württemberg;
                Award ID: Juniorprofessurenprogramm
                Award Recipient :
                Funded by: Boehringer Ingelheim Ulm University BioCenter (BIU)
                Award Recipient :
                Funded by: Boehringer Ingelheim Ulm University BioCenter (BIU)
                Award Recipient :
                This work was supported by the Deutsche Forschungsgemeinschaft (DFG) RO2173/3-1 (WR), RO2173/3-2 (WR), JU2859/2-1 (SJ) and the German Federal Ministry of Education and Research (BMBF) (e:Med-SYMBOL-HF grant #01ZX1407A) (SJ) as well as 01GS0108 (WR), 01GS0420 (WR), 01GS0836 (WR) and 01GS1104 (WR) (NGFNplus), and 01KU0901C (WR) Insight-DCM; the European Union (EU-Inheritance) (WR); the Ministerium für Wissenschaft, Forschung und Kunst Baden-Württemberg (MWK) Juniorprofessurenprogramm (SJ) and the Boehringer Ingelheim Ulm University BioCenter (BIU) (WR/SJ). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.
                Categories
                Research Article
                Research and Analysis Methods
                Model Organisms
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                Zebrafish
                Biology and Life Sciences
                Organisms
                Animals
                Vertebrates
                Fishes
                Osteichthyes
                Zebrafish
                Biology and Life Sciences
                Developmental Biology
                Embryology
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                Anatomy
                Cardiovascular Anatomy
                Heart
                Medicine and Health Sciences
                Anatomy
                Cardiovascular Anatomy
                Heart
                Biology and Life Sciences
                Molecular Biology
                Molecular Biology Techniques
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                Gene Expression and Vector Techniques
                Hyperexpression Techniques
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                Biology and Life Sciences
                Anatomy
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                Cardiac Ventricles
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