Phosphodiesterases Expression during Murine Cardiac Development

3′-5′ cyclic nucleotide phosphodiesterases (PDEs) are a large family of enzymes playing a fundamental role in the control of intracellular levels of cAMP and cGMP. Emerging evidence suggested an important role of phosphodiesterases in heart formation, but little is known about the expression of phosphodiesterases during cardiac development. In the present study, the pattern of expression and enzymatic activity of phosphodiesterases was investigated at different stages of heart formation. C57BL/6 mice were mated and embryos were collected from 14.5 to 18.5 days of development. Data obtained by qRT-PCR and Western blot analysis showed that seven different isoforms are expressed during heart development, and PDE1C, PDE2A, PDE4D, PDE5A and PDE8A are modulated from E14.5 to E18.5. In heart homogenates, the total cAMP and cGMP hydrolytic activity is constant at the evaluated times, and PDE4 accounts for the majority of the cAMP hydrolyzing ability and PDE2A accounts for cGMP hydrolysis. This study showed that a subset of PDEs is expressed in developing mice heart and some of them are modulated to maintain constant nucleotide phosphodiesterase activity in embryonic and fetal heart.

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Abstract 257: Clonal Analysis Of Multipotent Cardiovascular Progenitors That Generate Cardiomyocytes During Development

Circulation Research ◽

10.1161/res.115.suppl_1.257 ◽

2014 ◽

Vol 115 (suppl_1) ◽

Author(s):

Konstantina Ioanna Sereti ◽

Paniz Kamran Rashani ◽

Peng Zhao ◽

Reza Ardehali

Keyword(s):

Single Cell ◽

Heart Development ◽

Cardiac Development ◽

Clonal Analysis ◽

Cardiac Cells ◽

Single Cell Level ◽

Reporter System ◽

Cell Level ◽

Cardiac Progenitors

It has been proposed that cardiac development in lower vertebrates is driven by the proliferation of cardiomyocytes. Similarly, cycling myocytes have been suggested to direct cardiac regeneration in neonatal mice after injury. Although, the role of cardiomyocyte proliferation in cardiac tissue generation during development has been well documented, the extent of this contribution as well as the role of other cell types, such as progenitor cells, still remains controversial. Here we used a novel stochastic four-color Cre-dependent reporter system (Rainbow) that allows labeling at a single cell level and retrospective analysis of the progeny. Cardiac progenitors expressing Mesp1 or Nkx2.5 were shown to be a source of cardiomyocytes during embryonic development while the onset of αMHC expression marked the developmental stage where the capacity of cardiac cells to proliferate diminishes significantly. Through direct clonal analysis we provide strong evidence supporting that cardiac progenitors, as opposed to mature cardiomyocytes, are the main source of cardiomyocytes during cardiac development. Moreover, we have identified quadri-, tri-, bi, and uni-potent progenitors that at a single cell level can generate cardiomyocytes, fibroblasts, endothelial and smooth muscle cells. Although existing cardiomyocytes undergo limited proliferation, our data indicates that it is mainly the progenitors that contribute to heart development. Furthermore, we show that the limited proliferation capacity of cardiomyocytes observed during normal development was enhanced following neonatal cardiac injury allowing almost complete regeneration of the scared tissue. However, this ability was largely absent in adult injured hearts. Detailed characterization of dividing cardiomyocytes and proliferating progenitors would greatly benefit the development of novel therapeutic options for cardiovascular diseases.

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Abstract 052: Epicardial-derived Adrenomedullin Drives Cardiac Hyperplasia During Embryogenesis

Circulation Research ◽

10.1161/res.113.suppl_1.a052 ◽

2013 ◽

Vol 113 (suppl_1) ◽

Author(s):

Sarah E Wetzel-Strong ◽

Manyu Li ◽

Toshio Nishikimi ◽

Kathleen M Caron

Keyword(s):

Embryonic Development ◽

Heart Development ◽

Cardiac Development ◽

Lymphatic Vessels ◽

Heart Weight ◽

Cardiovascular Development ◽

Proliferating Cells ◽

Over Expression ◽

Knockout Animals

The multi-functional peptide adrenomedullin ( Adm = gene, AM = protein) plays important roles in embryonic development and disease. Previous studies demonstrated that Adm knockout mice die at embryonic day 13.5 with small, disorganized hearts and hypoplastic lymphatic vessels, highlighting the importance of this peptide in normal cardiovascular development. Since Adm knockout animals are embryonic lethal, our goal was to generate and characterize a novel model of Adm over-expression to study the role of Adm during development and disease processes. Through gene targeting techniques, we generated a novel mouse model of Adm over-expression, abbreviated as Adm hi/hi . When we assessed gene expression of Adm from 10 different tissues, we found Adm hi/hi mice express 3- to 15-fold more Adm than wildtype littermates. Additionally, peptide levels of AM in lung and kidney, as well as circulating plasma levels of AM were elevated 3-fold over wildtype mice, indicating a functional increase in AM. Our initial analysis revealed that adult Adm hi/hi mice have larger heart weight to body weight ratios than wildtype littermates (4.93±0.23 vs. 5.96±0.29, n = 11-12). We found that compared to wildtype, Adm hi/hi embryos have more proliferating cells during heart development (14.46±1.11 vs. 31.97±2.84, n=4), indicating that hyperplasia drives Adm hi/hi heart enlargement. By crossing the Adm hi/hi line to different tissue-specific Cre lines, we were able to excise the stabilizing bovine growth hormone 3’UTR, thereby returning Adm expression levels back to wildtype in cells with active Cre recombinase. Using this approach, we identified the epicardium as a major source of AM during cardiac development. In conclusion, we found that AM derived primarily from the epicardium drives cardiac hyperplasia during embryonic development resulting in persistent, enlarged hearts of adult Adm hi/hi mice. Since our Adm hi/hi mice recapitulate the 3-fold plasma elevation of AM observed during human disease, this mouse line will be a useful tool for studying the role of elevated AM during disease.

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The zebrafish as a model for cardiac development and regeneration

10.1093/med/9780198757269.003.0029 ◽

2018 ◽

Cited By ~ 1

Author(s):

Bill Chaudhry ◽

José Luis de la Pompa ◽

Nadia Mercader

Keyword(s):

Heart Development ◽

Cardiac Development ◽

Model Organism ◽

Laboratory Model ◽

Developmental Studies ◽

Sequencing Technologies ◽

Technological Advances ◽

The Common ◽

Zebrafish Heart

The zebrafish has become an established laboratory model for developmental studies and is increasingly used to model aspects of human development and disease. However, reviewers and grant funding bodies continue to speculate on the utility of this Himalayan minnow. In this chapter we explain the similarities and differences between the heart from this distantly related vertebrate and the mammalian heart, in order to reveal the common fundamental processes and to prevent misleading extrapolations. We provide an overview of zebrafish including their husbandry, development, peculiarities of their genome, and technological advances, which make them a highly tractable laboratory model for heart development and disease. We discuss the controversies around morphants and mutants, and relate the development and structures of the zebrafish heart to mammalian counterparts. Finally, we give an overview of regeneration in the zebrafish heart and speculate on the role of the model organism in next-generation sequencing technologies.

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The Role of Shear Stress on ET-1, KLF2, and NOS-3 Expression in the Developing Cardiovascular System of Chicken Embryos in a Venous Ligation Model

Physiology ◽

10.1152/physiol.00023.2007 ◽

2007 ◽

Vol 22 (6) ◽

pp. 380-389 ◽

Cited By ~ 59

Author(s):

Bianca C. W. Groenendijk ◽

Kim Van der Heiden ◽

Beerend P. Hierck ◽

Robert E. Poelmann

Keyword(s):

Shear Stress ◽

Heart Development ◽

Cardiac Development ◽

Background Information ◽

Endothelial Shear Stress ◽

Oxide Synthase ◽

Endothelial Nitric Oxide ◽

Cardiovascular Anomalies ◽

Chicken Model

In this review, the role of wall shear stress in the chicken embryonic heart is analyzed to determine its effect on cardiac development through regulating gene expression. Therefore, background information is provided for fluid dynamics, normal chicken and human heart development, cardiac malformations, cardiac and vitelline blood flow, and a chicken model to induce cardiovascular anomalies. A set of endothelial shear stress-responsive genes coding for endothelin-1 (ET-1), lung Krüppel-like factor (LKLF/KLF2), and endothelial nitric oxide synthase (eNOS/NOS-3) are active in development and are specifically addressed.

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GATA factors in vertebrate heart development and disease

Expert Reviews in Molecular Medicine ◽

10.1017/s1462399406000093 ◽

2006 ◽

Vol 8 (22) ◽

pp. 1-20 ◽

Cited By ~ 82

Author(s):

Alison Brewer ◽

John Pizzey

Keyword(s):

Transcription Factors ◽

Heart Development ◽

Regulatory Networks ◽

Cardiac Development ◽

Cellular Mechanisms ◽

Cardiac Morphogenesis ◽

Functional Roles ◽

Gata Factors ◽

Evolutionarily Conserved ◽

Heart Formation

Vertebrate heart formation is dependent upon complex hierarchical gene regulatory networks, which effect both the specification and differentiation of cardiomyocytes and subsequently cardiac morphogenesis. GATA-4, -5 and -6 comprise an evolutionarily conserved subfamily of transcription factors, which are expressed within the precardiac mesoderm from early stages in its specification and continue to be expressed within the adult heart. We review here the functional roles of individual GATA transcription factors in cardiac development, normal homeostasis and disease. We also review the cellular mechanisms employed to regulate the expression and downstream targets of the different GATA factors.

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The role of KMT2D and KDM6A in cardiac development: A cross-species analysis in humans, mice, and zebrafish

10.1101/2020.04.03.024646 ◽

2020 ◽

Author(s):

Rwik Sen ◽

Ezra Lencer ◽

Elizabeth A. Geiger ◽

Kenneth L. Jones ◽

Tamim H. Shaikh ◽

...

Keyword(s):

Gene Expression ◽

Heart Development ◽

Target Genes ◽

Cardiac Development ◽

Heart Defects ◽

Neural Crest Cell ◽

Kabuki Syndrome ◽

Rna Seq ◽

Wide Range

AbstractCongenital Heart Defects (CHDs) are the most common form of birth defects, observed in 4-10/1000 live births. CHDs result in a wide range of structural and functional abnormalities of the heart which significantly affect quality of life and mortality. CHDs are often seen in patients with mutations in epigenetic regulators of gene expression, like the genes implicated in Kabuki syndrome – KMT2D and KDM6A, which play important roles in normal heart development and function. Here, we examined the role of two epigenetic histone modifying enzymes, KMT2D and KDM6A, in the expression of genes associated with early heart and neural crest cell (NCC) development. Using CRISPR/Cas9 mediated mutagenesis of kmt2d, kdm6a and kdm6al in zebrafish, we show cardiac and NCC gene expression is reduced, which correspond to affected cardiac morphology and reduced heart rates. To translate our results to a human pathophysiological context and compare transcriptomic targets of KMT2D and KDM6A across species, we performed RNA sequencing (seq) of lymphoblastoid cells from Kabuki Syndrome patients carrying mutations in KMT2D and KDM6A. We compared the human RNA-seq datasets with RNA-seq datasets obtained from mouse and zebrafish. Our comparative interspecies analysis revealed common targets of KMT2D and KDM6A, which are shared between species, and these target genes are reduced in expression in the zebrafish mutants. Taken together, our results show that KMT2D and KDM6A regulate common and unique genes across humans, mice, and zebrafish for early cardiac and overall development that can contribute to the understanding of epigenetic dysregulation in CHDs.

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The role of the neural crest in cardiac development

10.1093/med/9780198757269.003.0019 ◽

2018 ◽

Author(s):

Laura A. Dyer ◽

Margaret L. Kirby

Keyword(s):

Neural Crest ◽

Neural Tube ◽

Heart Development ◽

Cardiac Development ◽

Signalling Pathways ◽

Pharyngeal Arches ◽

Cardiac Neural Crest ◽

Final Destination ◽

Dorsal Neural Tube

The cardiac neural crest (CNC) plays pivotal roles in numerous steps of cardiac development. Every aspect of the CNC cell’s lifespan is highly orchestrated, from its induction in the dorsal neural tube to its migration to its differentiation at its final destination. During migration, CNC cells are affected by their environment and simultaneously modulate the extra-cellular milieu through which they migrate. In the pharyngeal arches, CNC cells repattern the originally symmetrical arch arteries, producing the great arteries. Because the cardiac neural crest is essential for many aspects of heart development, it is unsurprising that human CNC-related syndromes have severe phenotypes. This chapter describes how CNC cells are formed and contribute to their final destinations. Essential signalling pathways are presented in the context of CNC development, and CNC-related syndromes are included to highlight this population’s broad importance during development.

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Cardiac Development: A Glimpse on Its Translational Contributions

Hearts ◽

10.3390/hearts2010008 ◽

2021 ◽

Vol 2 (1) ◽

pp. 87-118

Author(s):

Diego Franco ◽

Carlos Garcia-Padilla ◽

Jorge N. Dominguez ◽

Estefania Lozano-Velasco ◽

Amelia Aranega

Keyword(s):

Fetal Heart ◽

Developmental Process ◽

Cardiac Development ◽

Conduction System ◽

Molecular Pathways ◽

Straight Tube ◽

Cardiac Pathology ◽

Heart Formation ◽

Left Right Asymmetry ◽

The Right

Cardiac development is a complex developmental process that is initiated soon after gastrulation, as two sets of precardiac mesodermal precursors are symmetrically located and subsequently fused at the embryonic midline forming the cardiac straight tube. Thereafter, the cardiac straight tube invariably bends to the right, configuring the first sign of morphological left–right asymmetry and soon thereafter the atrial and ventricular chambers are formed, expanded and progressively septated. As a consequence of all these morphogenetic processes, the fetal heart acquired a four-chambered structure having distinct inlet and outlet connections and a specialized conduction system capable of directing the electrical impulse within the fully formed heart. Over the last decades, our understanding of the morphogenetic, cellular, and molecular pathways involved in cardiac development has exponentially grown. Multiples aspects of the initial discoveries during heart formation has served as guiding tools to understand the etiology of cardiac congenital anomalies and adult cardiac pathology, as well as to enlighten novels approaches to heal the damaged heart. In this review we provide an overview of the complex cellular and molecular pathways driving heart morphogenesis and how those discoveries have provided new roads into the genetic, clinical and therapeutic management of the diseased hearts.

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Building and Repairing the Heart: What Can We Learn from Embryonic Development?

BioMed Research International ◽

10.1155/2014/679168 ◽

2014 ◽

Vol 2014 ◽

pp. 1-8 ◽

Cited By ~ 6

Author(s):

Ana G. Freire ◽

Tatiana P. Resende ◽

Perpétua Pinto-do-Ó

Keyword(s):

Heart Development ◽

Cardiac Development ◽

Embryonic Stem ◽

Cardiac Pathophysiology ◽

Distinct Cell ◽

Cell Sources ◽

Cardiac Lineage ◽

Heart Formation ◽

Tissue Recovery

Mammalian heart formation is a complex morphogenetic event that depends on the correct temporal and spatial contribution of distinct cell sources. During cardiac formation, cellular specification, differentiation, and rearrangement are tightly regulated by an intricate signaling network. Over the last years, many aspects of this network have been uncovered not only due to advances in cardiac development comprehension but also due to the use of embryonic stem cells (ESCs)in vitromodel system. Additionally, several of these pathways have been shown to be functional or reactivated in the setting of cardiac disease. Knowledge withdrawn from studying heart development, ESCs differentiation, and cardiac pathophysiology may be helpful to envisage new strategies for improved cardiac repair/regeneration. In this review, we provide a comparative synopsis of the major signaling pathways required for cardiac lineage commitment in the embryo and murine ESCs. The involvement and possible reactivation of these pathways following heart injury and their role in tissue recovery will also be discussed.

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CFTR deficiency causes cardiac dysplasia during zebrafish embryogenesis and is associated with dilated cardiomyopathy

10.21203/rs.3.rs-18648/v1 ◽

2020 ◽

Author(s):

Yanyan Liu ◽

Ziyuan Lin ◽

Mingfeng Liu ◽

Huijuan Liao ◽

Yan Chen ◽

...

Keyword(s):

Heart Disease ◽

Dilated Cardiomyopathy ◽

Heart Development ◽

Cardiac Development ◽

Myocardial Dysfunction ◽

Primary Defect ◽

Zebrafish Model ◽

Human Heart Failure ◽

Stage Of Development

Abstract Background: Mutations in the CFTR gene cause cystic fibrosis (CF) with myocardial dysfunction. However, it remains unknown whether CF-related heart disease is a secondary effect of pulmonary disease, or an intrinsic primary defect in the CF heart. Results: Here, we used a zebrafish model, which lacks lung tissue, to investigate the role of CFTR in cardiogenesis during embryonic development. Our findings demonstrate that a loss of CFTR impairs cardiac development from the cardiac progenitor stage of heart development, resulting in cardiac looping defects, a dilated atrium, pericardial edema, and a decrease in heart rate. Furthermore, we found that cardiac development is perturbed in wild type embryos treated with a gating speciﬁc Cftr channel inhibitor, CFTRinh-172, at the blastula stage of development, but not with treatment at later stages. Gene expression analysis of blastulas indicated that transcript levels, including mRNAs associated with cardiovascular diseases, were significantly altered in embryos derived from cftr mutants relative to controls. To evaluate the role of CFTR in human heart failure, we performed a genetic association study on individuals with dilated cardiomyopathy and found that CFTR containing I556V mutation, which causes a channel defect, is associated with the disease. Similar to well-studied channel-defective CFTR mutants, CFTR I556V mRNA failed to restore cardiac dysplasia in mutant embryos. Conclusions: The present study reveals an important role for the CFTR ion channel in regulating cardiac development during early embryogenesis, supporting the hypothesis that CF-related heart disease results from an intrinsic primary defect in the CF heart.

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