Lessons from the lily pad: using Xenopus to understand heart disease

Bartlett, Heather L.; Weeks, Daniel L.

doi:10.1016/j.ddmod.2009.02.006

Cited by 8 publications

(8 citation statements)

References 26 publications

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“…Molecular mechanisms of heart development in the frog Xenopus are highly conserved with those of mouse and human (Brown et al, 2003; Gormley and Nascone-Yoder, 2003;Mohun et al, 2003;Brown et al, 2005; Garriock et al, 2005; Goetz et al, 2006;Bartlett et al, 2007; Langdon et al, 2007;Warkman and Krieg, 2007;Bartlett and Weeks, 2008;Afouda and Hoppler, 2009;Evans et al, 2010;Mandel et al, 2010; Kaltenbrun et al, 2011; Langdon et al, 2012). We have confirmed that, similar to other vertebrates, Xenopus epicardium develops as a specialized layer of cells surrounding the heart and is derived from a mesothelial precursor structure, the PEO, located on the ST (Jahr et al, 2008).…”

Section: Conserved Epicardial Development In Xenopusmentioning

confidence: 99%

Tcf21 regulates the specification and maturation of proepicardial cells

Tandon

Miteva

et al. 2013

Development

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SUMMARYThe epicardium is a mesothelial cell layer essential for vertebrate heart development and pertinent for cardiac repair post-injury in the adult. The epicardium initially forms from a dynamic precursor structure, the proepicardial organ, from which cells migrate onto the heart surface. During the initial stage of epicardial development crucial epicardial-derived cell lineages are thought to be determined. Here, we define an essential requirement for transcription factor Tcf21 during early stages of epicardial development in Xenopus, and show that depletion of Tcf21 results in a disruption in proepicardial cell specification and failure to form a mature epithelial epicardium. Using a mass spectrometry-based approach we defined Tcf21 interactions and established its association with proteins that function as transcriptional co-repressors. Furthermore, using an in vivo systems-based approach, we identified a panel of previously unreported proepicardial precursor genes that are persistently expressed in the epicardial layer upon Tcf21 depletion, thereby confirming a primary role for Tcf21 in the correct determination of the proepicardial lineage. Collectively, these studies lead us to propose that Tcf21 functions as a transcriptional repressor to regulate proepicardial cell specification and the correct formation of a mature epithelial epicardium. KEY WORDS: Tcf21, Proepicardial organ, Heart developmentTcf21 regulates the specification and maturation of proepicardial cells ZnCl 2 , 1 μM CaCl 2 , 0.5% Triton X-100 (v/v), 150 mM NaCl, 4 μg/ml DNase, 1/100 (v/v) Protease Inhibitor Cocktail and 1/100 Phosphatase Inhibitor Cocktail] using 5 ml lysis buffer/g cell powder. Lysates were homogenized, subjected to centrifugation, and supernatants incubated for 1 hour with 7 mg magnetic beads (M270 Epoxy Dynabeads, Invitrogen) conjugated with anti-EGFP antibodies (Cristea et al., 2005). Proteins were eluted by incubation for 10 minutes (70°C) in 30 µl 1× LDS sample buffer (Invitrogen) containing 1× Reducing Agent (Invitrogen), followed by shaking at room temperature for 10 minutes. Panna In-solution digestion, mass spectrometry analysis and data processingProtein IP eluates were prepared as described Greco et al., 2011;Tsai et al., 2012). Briefly, IP eluates were mixed with 8 M urea in aqueous 0.1 M Tris-HCl pH 8.0, applied to ultrafiltration Vivacon 500 units (Sartorius Stedim), and centrifuged at 14,000 g for 40 minutes at 20°C. Samples were washed, alkylated and digested with trypsin (Promega) overnight at 37°C. Resulting peptides were collected by centrifugation, acidified with trifluoroacetic acid, concentrated by vacuum centrifugation, and desalted using Empore C18 StageTips (Rappsilber et al., 2007;. Peptides were analyzed by nLC-MS/MS using a Dionex Ultimate 3000 RSLC system coupled online to an LTQ-Orbitrap Velos mass spectrometer (ThermoFisher Scientific) Tsai et al., 2012). Peptides were fragmented by collisioninduced dissociation (CID) and the MS/MS spectra were extracted by Proteome Discoverer (ThermoFishe...

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Section: Conserved Epicardial Development In Xenopusmentioning

confidence: 99%

Tcf21 regulates the specification and maturation of proepicardial cells

Tandon

Miteva

et al. 2013

Development

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“…It is well established that the molecular mechanisms of early heart development are highly conserved from Xenopus to human (Brade et al, 2007;Goetz and Conlon, 2007;Warkman and Krieg, 2007;Bartlett and Weeks, 2008;Afouda and Hoppler, 2009;Gessert and Kuhl, 2009;Evans et al, 2010;Kaltenbrun et al, 2011;Tandon et al, 2013), and studies in Xenopus have demonstrated an essential role for the transcription factor CASZ1 in cardiac differentiation and heart morphogenesis (Christine and Conlon, 2008). To ascertain the molecular mechanism by which CASZ1 functions in cardiac development, we undertook a yeast two-hybrid screen, using fulllength Xenopus CASZ1 as bait to screen a cDNA library that we generated from stage 28 Xenopus cardiac enriched tissue, a period that approximately corresponds to E7.5 in mouse and 18±1 days in human.…”

Section: Casz1 Interacts With Chd5mentioning

confidence: 99%

Congenital heart disease protein 5 associates with CASZ1 to maintain myocardial tissue integrity

et al. 2014

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The identification and characterization of the cellular and molecular pathways involved in the differentiation and morphogenesis of specific cell types of the developing heart are crucial to understanding the process of cardiac development and the pathology associated with human congenital heart disease. Here, we show that the cardiac transcription factor CASTOR (CASZ1) directly interacts with congenital heart disease 5 protein (CHD5), which is also known as tryptophanrich basic protein (WRB), a gene located on chromosome 21 in the proposed region responsible for congenital heart disease in individuals with Down's syndrome. We demonstrate that loss of CHD5 in Xenopus leads to compromised myocardial integrity, improper deposition of basement membrane, and a resultant failure of hearts to undergo cell movements associated with cardiac formation. We further report that CHD5 is essential for CASZ1 function and that the CHD5-CASZ1 interaction is necessary for cardiac morphogenesis. Collectively, these results establish a role for CHD5 and CASZ1 in the early stages of vertebrate cardiac development.

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“…The most extensively used of these techniques are the use of morpholino oligonucleotides (MOs), which inhibit the function of specific genes by preventing translation or splicing of messenger RNA (mRNA). This technique has resulted in the publication of many studies of heart development in Xenopus that have advanced our understanding of this process in vertebrates (Bartlett and Weeks, 2008; Brown and others, 2007; Brown and others, 2005; Christine and Conlon, 2008; Garriock and others, 2005a; Hilton and others, 2007; Inui and others, 2006; Kumano and others, 2006; Movassagh and Philpott, 2008; Nagao and others, 2008; Small and others, 2005; Zhang and others, 2005). MO antisense oligonucleotides are neutrally charged synthetic nucleic acid analogs that are stable, soluble, and bind to RNA with high affinity (Heasman and others, 2000; Moulton, 2007).…”

Section: Methods For Studying Heart Development and Disease In Xenopusmentioning

confidence: 99%

“…Moreover, recent sequence annotation and assembly of the Xenopus tropicalis genome has demonstrated that it has long regions in which genes exhibit remarkably similar synteny relationships to those found in the human genome (Hellsten and others, 2010; Showell and Conlon, 2007). Specifically, regarding human congenital heart disease (CHD), Xenopus has unique advantages for studying cardiovascular development (Bartlett and Weeks, 2008; Evans and others, 2010; Warkman and Krieg, 2006). First, early Xenopus development can proceed in the absence of a functional circulation system, allowing defects to be extensively analyzed in living embryos.…”

Section: Introductionmentioning

confidence: 99%

Xenopus: An emerging model for studying congenital heart disease

Kaltenbrun

Tandon

Amin

et al. 2011

Birth Defects Research

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Congenital heart defects affect nearly 1% of all newborns and are a significant cause of infant death. Clinical studies have identified a number of congenital heart syndromes associated with mutations in genes that are involved in the complex process of cardiogenesis. The African clawed frog, Xenopus, has been instrumental in studies of vertebrate heart development and provides a valuable tool to investigate the molecular mechanisms underlying human congenital heart diseases. In this review, we discuss the methodologies that make Xenopus an ideal model system to investigate heart development and disease. We also outline congenital heart conditions linked to cardiac genes that have been well-studied in Xenopus and describe some emerging technologies that will further aid in the study of these complex syndromes.

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Lessons from the lily pad: using Xenopus to understand heart disease

Cited by 8 publications

References 26 publications

Tcf21 regulates the specification and maturation of proepicardial cells

Tcf21 regulates the specification and maturation of proepicardial cells

Congenital heart disease protein 5 associates with CASZ1 to maintain myocardial tissue integrity

Xenopus: An emerging model for studying congenital heart disease

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