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1.
PLoS One ; 15(7): e0234069, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32649674

RESUMO

Recent discoveries of at least two heart fields and dynamic nature of cardiac development as well as controversies regarding the participation of heart fields in development of different heart structures led us to investigate the dynamics of incorporation of the first and second heart fields and prospective fate of the straight heart tube by labeling chicken embryos in vivo with the fluorescent lipophilic dye DiI. The cephalic and caudal limits of the anterior and posterior segments of the straight heart tube were labeled in two groups of embryos. Labels were tracked along the "C," "S," and "U" loops up to the tetracavitary or mature heart (n = 30 embryos/group; torsion and looping stage). To determine whether the atria and atrioventricular canal are derived from the first heart field the straight heart tube was cultured in vitro and immunodetection of Sox-9 and troponin I was performed to identify the mesenchymal and myocardial lineages respectively. Proliferating cell nuclear antigen (PCNA) immunodetection was used to determine the involvement of cell proliferation in heart tube development during torsion and looping. Embryological constitution of the straight heart tube and heart looping (C, S, and U) were not consistent with current descriptions. In fact, right ventricle precursors were absent in the straight heart tube derived from the first heart field. During torsion and looping, the cephalic segment of the straight heart tube gradually shifted into the heart tube until it was located at the myocardial interventricular septum in the tetracavitary heart. In contrast, the caudal segment of the straight heart tube was elongated and remodeled to become the first heart field derived left ventricle and the proximal part of the ventricular inlets. The ventricular outflows, right ventricle, distal part of the ventricular inlets, and atria developed from the second heart field.


Assuntos
Coração/embriologia , Animais , Carbocianinas , Divisão Celular , Linhagem da Célula , Embrião de Galinha , Corantes Fluorescentes , Mesoderma/citologia , Microscopia Eletrônica de Varredura , Miocárdio/química , Miocárdio/ultraestrutura , Organogênese , Antígeno Nuclear de Célula em Proliferação/análise , Fatores de Transcrição SOX9/análise , Troponina I/análise
2.
Ecotoxicol Environ Saf ; 201: 110808, 2020 Sep 15.
Artigo em Inglês | MEDLINE | ID: mdl-32516676

RESUMO

Cyprodinil is a broad-spectrum pyrimidine amine fungicide that has been reportedly used worldwide. However, toxicity studies of cyprodinil on aquatic organisms, specifically zebrafish (Danio rerio), are lacking. In our present study, we predicted cyprodinil binding to the aryl hydrocarbon receptor (AhR) by using molecular docking simulation. Then, we used recombinant HepG2 cells and Tg(cyp1a1-12DRE:egfp) transgenic zebrafish to further assess the AhR agonistic activity of cyprodinil. Besides, the significant upregulation of cyp1a1 further verified that statement. Moreover, we found that zebrafish exposure to cyprodinil induced developmental toxicity in the larvae, particularly during cardiac development. The expression levels of cardiac development-related genes, namely tbx5, nkx2.5, gata4, and tnnt2, were markedly altered, which might cause the adverse effects of cyprodinil on cardiac function and development. In summary, we found that cyprodinil, as an AhR agonist, induced development toxicity in zebrafish larvae, especially on cardiac. Data here can assess the potential effects on organisms in the aquatic environment and promote the regulation and safe use of cyprodinil.


Assuntos
Animais Geneticamente Modificados/metabolismo , Larva/efeitos dos fármacos , Pirimidinas/toxicidade , Receptores de Hidrocarboneto Arílico/agonistas , Poluentes Químicos da Água/toxicidade , Peixe-Zebra/metabolismo , Animais , Citocromo P-450 CYP1A1/genética , Citocromo P-450 CYP1A1/metabolismo , Embrião não Mamífero/efeitos dos fármacos , Embrião não Mamífero/metabolismo , Fungicidas Industriais/metabolismo , Coração/efeitos dos fármacos , Coração/embriologia , Células Hep G2 , Humanos , Larva/genética , Larva/metabolismo , Simulação de Acoplamento Molecular , Organogênese/efeitos dos fármacos , Organogênese/genética , Ligação Proteica , Peixe-Zebra/genética
3.
Chemosphere ; 251: 126610, 2020 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-32443250

RESUMO

Trichloroethylene (TCE), a widely used organic solvent, is a common environmental pollutant. Increasing evidence indicates that maternal TCE exposure is associated with congenital cardiac defects, but the underlining mechanisms remain largely unknown. In this study, we revealed that TCE exposure significantly induced heart defects and dysfunctions in zebrafish embryos. Heart tissues were dissected and subjected to high throughput sequencing and qPCR to identify differentially expressed miRNAs and mRNAs. The effects of miRNA were further verified by microinjection of antagomir or agomir. Reactive Oxygen Species (ROS) and cell proliferation were measured by using dichlorodihydrofluorescein diacetate (DCFH-DA) and EdU staining, respectively. Our results showed that 19 miRNAs were downregulated whereas 48 miRNAs were upregulated in the heart of zebrafish embryos. The downregulation of miR-133a and the upregulation of miR-182 were further validated. Moreover, we found that miR-133a agomir significantly alleviated the TCE-induced heart defects while miR-133a antagomir mimicked the toxic effect of TCE on heart development. Furthermore, miR-133a agomir significantly counteracted TCE-induced ROS production and excessive cell proliferation in the heart of zebrafish embryos. In conclusion, our results indicate that miR-133a mediates TCE-induced ROS generation, leading to excessive cell proliferation and heart defects.


Assuntos
Desenvolvimento Embrionário/efeitos dos fármacos , Cardiopatias Congênitas/induzido quimicamente , Coração/efeitos dos fármacos , MicroRNAs/genética , Tricloroetileno/toxicidade , Poluentes Químicos da Água/toxicidade , Peixe-Zebra , Animais , Cardiotoxicidade , Proliferação de Células/efeitos dos fármacos , Proliferação de Células/genética , Regulação para Baixo , Embrião não Mamífero/efeitos dos fármacos , Desenvolvimento Embrionário/genética , Coração/embriologia , Cardiopatias Congênitas/genética , Regulação para Cima
4.
PLoS One ; 15(5): e0233719, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32469960

RESUMO

The formation of a tube-like structure is a basic step in the making of functional hearts in vertebrates and invertebrates and therefore, its understanding provides important information on heart development and function. In Drosophila, the cardiac tube originates from two bilateral rows of dorsally migrating cells. On meeting at the dorsal midline, coordinated changes in cell shape and adhesive properties transform the two sheets of cells into a linear tube. ECM and transmembrane proteins linked to the cytoskeleton play an important role during these dynamic processes. Here we characterize the requirement of Cbl-Associated Protein (CAP) in Drosophila heart formation. In embryos, CAP is expressed in late migrating cardioblasts and is located preferentially at their luminal and abluminal periphery. CAP mutations result in irregular cardioblast alignment and imprecisely controlled cardioblast numbers. Furthermore, CAP mutant embryos show a strongly reduced heart lumen and an aberrant shape of lumen forming cardioblasts. Analysis of double heterozygous animals reveals a genetic interaction of CAP with Integrin- and Talin-encoding genes. In post-embryonic stages, CAP closely colocalizes with Integrin near Z-bands and at cell-cell contact sites. CAP mutants exhibit a reduced contractility in larval hearts and show a locally disrupted morphology, which correlates with a reduced pumping efficiency. Our observations imply a function of CAP in linking Integrin signaling with the actin cytoskeleton. As a modulator of the cytoskeleton, CAP is involved in the establishment of proper cell shapes during cardioblast alignment and cardiac lumen formation in the Drosophila embryo. Furthermore, CAP is required for correct heart function throughout development.


Assuntos
Movimento Celular , Proteínas de Drosophila/metabolismo , Embrião não Mamífero/embriologia , Coração/embriologia , Proteínas de Transporte de Monossacarídeos/metabolismo , Contração Miocárdica , Organogênese , Animais , Proteínas de Drosophila/genética , Drosophila melanogaster , Proteínas de Transporte de Monossacarídeos/genética , Mutação
5.
BMC Bioinformatics ; 21(1): 161, 2020 Apr 29.
Artigo em Inglês | MEDLINE | ID: mdl-32349652

RESUMO

BACKGROUND: Technological developments in the emerging field of spatial transcriptomics have opened up an unexplored landscape where transcript information is put in a spatial context. Clustering commonly constitutes a central component in analyzing this type of data. However, deciding on the number of clusters to use and interpreting their relationships can be difficult. RESULTS: We introduce SpatialCPie, an R package designed to facilitate cluster evaluation for spatial transcriptomics data. SpatialCPie clusters the data at multiple resolutions. The results are visualized with pie charts that indicate the similarity between spatial regions and clusters and a cluster graph that shows the relationships between clusters at different resolutions. We demonstrate SpatialCPie on several publicly available datasets. CONCLUSIONS: SpatialCPie provides intuitive visualizations of cluster relationships when dealing with Spatial Transcriptomics data.


Assuntos
Software , Transcriptoma/genética , Análise por Conglomerados , Regulação da Expressão Gênica no Desenvolvimento , Coração/embriologia , Humanos
6.
Chemosphere ; 256: 127133, 2020 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-32454355

RESUMO

Atmospheric fine particulate matter exposure (PM2.5) can increase the incidence and mortality of heart disease, and raise the risk of fetal congenital heart defect, which have recently drawn much attention. In this study, C57BL/6 mice were exposed to PM2.5 (approximately equivalent to 174 µg/m3) by intratracheal instillation during the gestation. After birth, 10 weeks old offspring mice were divided into four groups: male exposed group (ME), female exposed group (FE), male control group (MC), female control group (FC). The pathological injury, pro-inflammatory cytokines, histone acetylation levels, and expressions of GATA-binding protein 4 (GATA4) and downstream genes were investigated. The results showed that exposure to PM2.5 in utero increased pathological damage and TNF-α and IL-6 levels in hearts of offspring mice, and effects in ME were more serious than FE. Notably, GATA4 protein levels in hearts in ME were significantly lower than that of MC, accompanied by down-regulation of histone acetyltransferase (HAT)-p300 and up-regulation of histone deacetylase-SIRT3. As GATA4 downstream genes, ratios of ß-MHC gene expression to α-MHC significantly raised in ME relative to the MC. Results of chromatin immunoprecipitation (ChIP)-qPCR assay found that binding levels of acetylated histone 3 lysine 9 (H3K9ac) in GATA4 promoter region in the hearts of ME or FE were markedly decreased compared with their corresponding control groups. It suggested that maternal exposure to PM2.5 may cause cardiac injury in the offspring, heart damage of male mice was worse than female mice, in which process HAT-p300, H3K9ac, transcription factor GATA4 may play an important regulation role.


Assuntos
Poluentes Atmosféricos/toxicidade , Fator de Transcrição GATA4/genética , Regulação da Expressão Gênica no Desenvolvimento/efeitos dos fármacos , Coração/efeitos dos fármacos , Histonas/metabolismo , Material Particulado/toxicidade , Efeitos Tardios da Exposição Pré-Natal/induzido quimicamente , Acetilação , Animais , Animais Recém-Nascidos , Regulação para Baixo , Proteína p300 Associada a E1A/metabolismo , Feminino , Coração/embriologia , Coração/crescimento & desenvolvimento , Histonas/genética , Humanos , Masculino , Exposição Materna/efeitos adversos , Camundongos , Camundongos Endogâmicos C57BL , Gravidez , Efeitos Tardios da Exposição Pré-Natal/metabolismo , Efeitos Tardios da Exposição Pré-Natal/patologia , Caracteres Sexuais , Regulação para Cima
7.
Ecotoxicol Environ Saf ; 200: 110772, 2020 Sep 01.
Artigo em Inglês | MEDLINE | ID: mdl-32464444

RESUMO

Recently two-dimensional nanomaterials, such as graphene and molybdenum disulfide (MoS2), have received much attention as adsorbent materials for the effective removal of organic contaminants. MoS2 is attracting attention, not only for its chemical-physical properties, but also for its wide availability in nature as a constituent of molybdenite. The aim of this investigation was to assess the effects of different MoS2 concentrations (5 × 10-1, 5 × 10-2 and 5 × 10-3 mg/ml) on the embryonated eggs of Gallus gallus domesticus, according to Beck method. We evaluated the toxic effect of the MoS2 powder purchased at Sigma-Aldrich indicated as "received" and MoS2 powder treated via mechanical milling indicated as "ball mille". Subsequently, the embryos were sacrificed at different times of embryonic development (11th, 15th and 19th day after incubation) in order to evaluate their embryotoxic and teratogenic effects. The alterations of the embryonic development were studied by morphological and immunohistochemical analysis of the tissues. The results obtained have shown the toxicity of both powders of MoS2 with a high percentage of deaths and growth delays. Moreover, the immunohistochemical analysis performed on several tissue sections showed a strong positivity to the anti-metallothionein1 antibody only for the erythrocytes.


Assuntos
Dissulfetos/química , Dissulfetos/toxicidade , Desenvolvimento Embrionário/efeitos dos fármacos , Molibdênio/química , Molibdênio/toxicidade , Nanopartículas/química , Nanopartículas/toxicidade , Animais , Embrião de Galinha , Relação Dose-Resposta a Droga , Grafite/química , Coração/efeitos dos fármacos , Coração/embriologia , Fígado/efeitos dos fármacos , Fígado/embriologia , Fígado/patologia , Pulmão/efeitos dos fármacos , Pulmão/embriologia , Pulmão/patologia , Tamanho da Partícula , Propriedades de Superfície
8.
Subcell Biochem ; 95: 119-149, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32297298

RESUMO

As the first organ to form and function in all vertebrates, the heart is crucial to development. Tightly-regulated levels of retinoic acid (RA) are critical for the establishment of the regulatory networks that drive normal cardiac development. Thus, the heart is an ideal organ to investigate RA signaling, with much work remaining to be done in this area. Herein, we highlight the role of RA signaling in vertebrate heart development and provide an overview of the field's inception, its current state, and in what directions it might progress so that it may yield fruitful insight for therapeutic applications within the domain of regenerative medicine.


Assuntos
Regulação da Expressão Gênica no Desenvolvimento , Coração/embriologia , Transdução de Sinais , Tretinoína/metabolismo , Animais , Humanos , Medicina Regenerativa , Vertebrados/embriologia , Vertebrados/genética
9.
Adv Exp Med Biol ; 1236: 189-223, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32304074

RESUMO

Congenital heart defects (CHDs) are among the most common human birth defects. However, the etiology of a large proportion of CHDs remains undefined. Studies identifying the molecular and cellular mechanisms that underlie cardiac development have been critical to elucidating the origin of CHDs. Building upon this knowledge to understand the pathogenesis of CHDs requires examining how genetic or environmental stress changes normal cardiac development. Due to strong molecular conservation to humans and unique technical advantages, studies using zebrafish have elucidated both fundamental principles of cardiac development and have been used to create cardiac disease models. In this chapter we examine the unique toolset available to zebrafish researchers and how those tools are used to interrogate the genetic and environmental contributions to CHDs.


Assuntos
Meio Ambiente , Cardiopatias Congênitas/etiologia , Cardiopatias Congênitas/genética , Peixe-Zebra/genética , Animais , Interação Gene-Ambiente , Coração/embriologia , Cardiopatias Congênitas/embriologia , Cardiopatias Congênitas/patologia , Humanos , Peixe-Zebra/embriologia
10.
PLoS Genet ; 16(4): e1008730, 2020 04.
Artigo em Inglês | MEDLINE | ID: mdl-32251422

RESUMO

O-linked N-acetylglucosamine (GlcNAc) transferase (OGT) is the only enzyme catalyzing O-GlcNAcylation. Although it has been shown that OGT plays an essential role in maintaining postnatal heart function, its role in heart development remains unknown. Here we showed that loss of OGT in early fetal cardiomyocytes led to multiple heart developmental defects including hypertrabeculation, biventricular dilation, atrial septal defects, ventricular septal defects, and defects in coronary vessel development. In addition, RNA sequencing revealed that Angiopoietin-1, required within cardiomyocytes for both myocardial and coronary vessel development, was dramatically downregulated in cardiomyocyte-specific OGT knockout mouse hearts. In conclusion, our data demonstrated that OGT plays an essential role in regulating heart development through activating expression of cardiomyocyte Angiopoietin-1.


Assuntos
Coração/embriologia , Miócitos Cardíacos/metabolismo , N-Acetilglucosaminiltransferases/metabolismo , Angiopoietina-1/genética , Angiopoietina-1/metabolismo , Animais , Células Cultivadas , Coração/fisiologia , Camundongos , Camundongos Endogâmicos C57BL , N-Acetilglucosaminiltransferases/genética
11.
J Vis Exp ; (157)2020 03 10.
Artigo em Inglês | MEDLINE | ID: mdl-32225147

RESUMO

Congenital heart defects (CHD) are the most common type of birth defect in humans, affecting up to 1% of all live births. However, the underlying causes for CHD are still poorly understood. The developing mouse constitutes a valuable model for the study of CHD, because cardiac developmental programs between mice and humans are highly conserved. The protocol describes in detail how to produce mouse embryos of the desired gestational stage, methods to isolate and preserve the heart for downstream processing, quantitative methods to identify common types of CHD by histology (e.g., ventricular septal defects, atrial septal defects, patent ductus arteriosus), and quantitative histomorphometry methods to measure common muscular compaction phenotypes. These methods articulate all the steps involved in sample preparation, collection, and analysis, allowing scientists to correctly and reproducibly measure CHD.


Assuntos
Embrião de Mamíferos/anormalidades , Embrião de Mamíferos/patologia , Cardiopatias Congênitas/embriologia , Cardiopatias Congênitas/patologia , Histocitoquímica/métodos , Animais , Feminino , Coração/embriologia , Humanos , Camundongos Endogâmicos C57BL , Miocárdio/patologia , Inclusão em Parafina , Fenótipo
12.
J Vis Exp ; (157)2020 03 10.
Artigo em Inglês | MEDLINE | ID: mdl-32225157

RESUMO

Here, we describe an in vitro culture assay to study coronary angiogenesis. Coronary vessels feed the heart muscle and are of clinical importance. Defects in these vessels represent severe health risks such as in atherosclerosis, which can lead to myocardial infarctions and heart failures in patients. Consequently, coronary artery disease is one of the leading causes of death worldwide. Despite its clinical importance, relatively little progress has been made on how to regenerate damaged coronary arteries. Nevertheless, recent progress has been made in understanding the cellular origin and differentiation pathways of coronary vessel development. The advent of tools and technologies that allow researchers to fluorescently label progenitor cells, follow their fate, and visualize progenies in vivo have been instrumental in understanding coronary vessel development. In vivo studies are valuable, but have limitations in terms of speed, accessibility, and flexibility in experimental design. Alternatively, accurate in vitro models of coronary angiogenesis can circumvent these limitations and allow researchers to interrogate important biological questions with speed and flexibility. The lack of appropriate in vitro model systems may have hindered the progress in understanding the cellular and molecular mechanisms of coronary vessel growth. Here, we describe an in vitro culture system to grow coronary vessels from the sinus venosus (SV) and endocardium (Endo), the two progenitor tissues from which many of the coronary vessels arise. We also confirmed that the cultures accurately recapitulate some of the known in vivo mechanisms. For instance, we show that the angiogenic sprouts in culture from SV downregulate COUP-TFII expression similar to what is observed in vivo. In addition, we show that VEGF-A, a well-known angiogenic factor in vivo, robustly stimulates angiogenesis from both the SV and Endo cultures. Collectively, we have devised an accurate in vitro culture model to study coronary angiogenesis.


Assuntos
Vasos Coronários/fisiologia , Modelos Biológicos , Neovascularização Fisiológica , Animais , Fator II de Transcrição COUP/metabolismo , Reprogramação Celular , Vasos Coronários/embriologia , Dissecação , Embrião de Mamíferos/irrigação sanguínea , Matriz Extracelular/metabolismo , Feminino , Coração/embriologia , Coração/fisiologia , Ventrículos do Coração/embriologia , Humanos , Processamento de Imagem Assistida por Computador , Masculino , Camundongos , Gravidez , Técnicas de Cultura de Tecidos , Fixação de Tecidos , Fator A de Crescimento do Endotélio Vascular/metabolismo
13.
Zhongguo Yi Xue Ke Xue Yuan Xue Bao ; 42(1): 96-102, 2020 Feb 28.
Artigo em Chinês | MEDLINE | ID: mdl-32131947

RESUMO

Cilia are protruding cell structures on the cell surface and are found in almost every type of cell.According to the different structures and quantity of tubulins,cilia can be divided into two categories:motor cilia and sensory cilia.Sensory cilia are also called non-motor cilia and primary cilia,due to the composition and number of tubulins.They are closely related to the development of internal organs and many human physiological activities.Recent studies have demonstrated that cilia are involved in regulating the formation of left and right symmetry of the heart structure,and eventually the heart develops into the left-right asymmetry structures.Since congenital heart diseases(CHD)are characterized by abnormalities in the spatial structure of the heart chamber and outflow tract,cilia may play an important role in the pathogenesis of CHD.Cilia,mainly through ciliary transduction signal pathways,regulate both the formation of left and right asymmetrical structures and the polarity and the migration of cells.Therefore,a clear understanding of the regulation mechanism of ciliary signaling pathway on heart development can provide new therapeutic targets and new ideas for the clinical treatment of CHD and may offer new target genes for prenatal screening of CHD.This article summarizes recent advances in the role of cilia in heart development and CHD pathogenesis and its mechanisms.


Assuntos
Cílios/fisiologia , Cardiopatias/congênito , Coração/embriologia , Transdução de Sinais , Humanos
14.
Am J Physiol Heart Circ Physiol ; 318(5): H1308-H1315, 2020 05 01.
Artigo em Inglês | MEDLINE | ID: mdl-32216613

RESUMO

Noncoding RNAs (ncRNAs) are broadly described as RNA molecules that are not translated into protein. The investigation of dysregulated ncRNAs in human diseases such as cancer, neurological, and cardiovascular diseases has been under way for well over a decade. Micro-RNAs and long noncoding RNAs (lncRNAs) are the best characterized ncRNAs. These ncRNAs can have profound effects on the regulation of gene expression during cardiac development and disease. Importantly, ncRNAs are significant regulators of gene expression in several congenital heart diseases and can positively or negatively impact cardiovascular development. In this review, we focus on literature involving micro-RNAs and lncRNAs in the context of pediatric cardiovascular diseases, preclinical models of heart failure, and cardiac development.


Assuntos
Regulação da Expressão Gênica no Desenvolvimento , Cardiopatias Congênitas/genética , Insuficiência Cardíaca/genética , MicroRNAs/genética , Miocárdio/metabolismo , RNA Longo não Codificante/genética , Animais , Coração/embriologia , Cardiopatias Congênitas/metabolismo , Cardiopatias Congênitas/patologia , Insuficiência Cardíaca/metabolismo , Insuficiência Cardíaca/patologia , Humanos , MicroRNAs/metabolismo , RNA Longo não Codificante/metabolismo
15.
Nat Cell Biol ; 22(3): 332-340, 2020 03.
Artigo em Inglês | MEDLINE | ID: mdl-32123336

RESUMO

Mapping of the holistic cell behaviours sculpting the four-chambered mammalian heart has been a goal or previous studies, but so far only success in transparent invertebrates and lower vertebrates with two-chambered hearts has been achieved. Using a live-imaging system comprising a customized vertical light-sheet microscope equipped with a mouse embryo culture module, a heartbeat-gated imaging strategy and a digital image processing framework, we realized volumetric imaging of developing mouse hearts at single-cell resolution and with uninterrupted cell lineages for up to 1.5 d. Four-dimensional landscapes of Nppa+ cardiomyocyte cell behaviours revealed a blueprint for ventricle chamber formation by which biased outward migration of the outermost cardiomyocytes is coupled with cell intercalation and horizontal division. The inner-muscle architecture of trabeculae was developed through dual mechanisms: early fate segregation and transmural cell arrangement involving both oriented cell division and directional migration. Thus, live-imaging reconstruction of uninterrupted cell lineages affords a transformative means for deciphering mammalian organogenesis.


Assuntos
Ventrículos do Coração/citologia , Ventrículos do Coração/embriologia , Processamento de Imagem Assistida por Computador/métodos , Miócitos Cardíacos/citologia , Animais , Divisão Celular , Linhagem da Célula , Movimento Celular , Embrião de Mamíferos/citologia , Desenvolvimento Embrionário , Coração/embriologia , Camundongos , Microscopia , Morfogênese , Miocárdio/citologia , Análise de Célula Única , Técnicas de Cultura de Tecidos
16.
Environ Sci Process Impacts ; 22(3): 824-832, 2020 Mar 01.
Artigo em Inglês | MEDLINE | ID: mdl-32159184

RESUMO

In exploration of congenital heart defects produced by TCE, Hepatocyte Nuclear Factor 4 alpha (HNF4a) transcriptional activity was identified as a central component. TCE exposure altered gene transcription in the chick heart in a non-monotonic pattern where only low dose exposure inhibited transcription by HNF4a. As the chick embryo is non-placental, we examine here HNF4a as a target of TCE in developing mouse embryos. Benfluorex and Bi6015, published agonist and antagonist, respectively, of HNF4a were compared to low dose TCE exposure. Pregnant mice were exposed to 10 ppb (76 nM) TCE, 5 µM Benfluorex, 5 µM Bi6015, or a combination of Bi6015 and TCE in drinking water. Litters (E12) were collected during a sensitive window in heart development. Embryonic hearts were collected, pooled for extraction of RNA and marker expression was examined by quantitative PCR. Multiple markers, previously identified as sensitive to TCE exposure in chicks or as published targets of HNF4a transcription were significantly affected by Benfluorex, Bi6015 and TCE. Activity of TCE and both HNF4a-specific reagents on transcription argues that HNF4a is a component of TCE cardiotoxicity and likely a proximal target of low dose exposure during development. The effectiveness of these reagents after delivery in maternal drinking water suggests that neither maternal metabolism, nor placental transport is protective of exposure.


Assuntos
Tricloroetileno/toxicidade , Animais , Feminino , Coração/embriologia , Fator 4 Nuclear de Hepatócito/genética , Camundongos , Gravidez
17.
Cell Prolif ; 53(3): e12764, 2020 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-32077168

RESUMO

OBJECTIVES: microRNA-29 (miR-29) family have shown different expression patterns in cardiovascular diseases. Our study aims to explore the effect and mechanism of miR-29 family on cardiac development. MATERIALS AND METHODS: A total of 13 patients with congenital heart disease (CHD) and 7 controls were included in our study. Tissues were obtained from the right ventricular outflow tract (RVOT) after surgical resection or autopsy. The next-generation sequencing was applied to screen the microRNA expression profiles of CHD. Quantitative RT-PCR and Western blot were employed to measure genes expression. Tg Cmlc2: GFP reporter zebrafish embryos were injected with microRNA (miRNA) to explore its role in cardiac development in vivo. Dual-luciferase reporter assay was designed to validate the target gene of miRNAs. CCK-8 and EdU incorporation assays were performed to evaluate cardiomyocyte proliferation. RESULTS: Our study showed miR-29b-3p expression was significantly increased in the RVOT of the CHD patients. Injection of miR-29b-3p into zebrafish embryos induced higher mortality and malformation rates, developmental delay, cardiac malformation and dysfunction. miR-29b-3p inhibited cardiomyocyte proliferation, and its inhibitor promoted cardiomyocyte proliferation in vitro and in vivo. Furthermore, we identified that miR-29b-3p influenced cardiomyocyte proliferation by targeting NOTCH2, which was down-regulated in the RVOT of the CHD patients. CONCLUSION: This study reveals that miR-29b-3p functions as a novel regulator of cardiac development and inhibits cardiomyocyte proliferation via NOTCH2, which provides novel insights into the aetiology and potential treatment of CHD.


Assuntos
Cardiopatias Congênitas/genética , MicroRNAs/genética , Miócitos Cardíacos/patologia , Receptor Notch2/genética , Animais , Proliferação de Células , Células Cultivadas , Pré-Escolar , Regulação para Baixo , Feminino , Coração/embriologia , Cardiopatias Congênitas/patologia , Humanos , Lactente , Masculino , Camundongos , Miócitos Cardíacos/citologia , Miócitos Cardíacos/metabolismo , Regulação para Cima , Peixe-Zebra
18.
Aquat Toxicol ; 222: 105451, 2020 May.
Artigo em Inglês | MEDLINE | ID: mdl-32097808

RESUMO

Musk compounds are often used as to treat heart-related diseases and are widely used in Asia. Muscone is one of the most important physiologically active compounds of natural musk. Muscone is a chiral compound and can be further classified into S-muscone and R-muscone and both are present in synthetic musk. While these two chiral isomers have significant differences in odor properties, their difference in toxicity is still unknown. This study used zebrafish as an animal model to compare cardiac toxicities of S-muscone and R-muscone. Results showed that both compounds were acutely toxic to zebrafish embryos causing mortality, decreased hatching rate, pericardial edema, and decreased heart beat rate. These toxicities were modulated through increased Myh6 and Myh7 mRNA expression, and decreased thyroid genes (Trh, Thrß, and Dio3) expression. R-muscone caused higher toxicity than S-muscone at the same concentration. For safety, the chiral isomer composition of synthetic muscone should be carefully regulated in the future.


Assuntos
Cicloparafinas/toxicidade , Embrião não Mamífero/efeitos dos fármacos , Coração/efeitos dos fármacos , Poluentes Químicos da Água/toxicidade , Animais , Cardiotoxicidade , Cicloparafinas/química , Desenvolvimento Embrionário/efeitos dos fármacos , Coração/embriologia , Frequência Cardíaca/efeitos dos fármacos , Estereoisomerismo , Poluentes Químicos da Água/química , Peixe-Zebra/embriologia
19.
J Vis Exp ; (156)2020 02 06.
Artigo em Inglês | MEDLINE | ID: mdl-32090990

RESUMO

Zebrafish are increasingly utilized as a model organism for cardiomyopathies and regeneration. Current methods evaluating cardiac function fail to reliably detect segmental mechanics and are not readily feasible in zebrafish. Here we present a semiautomated, open-source method for the quantitative assessment of four-dimensional (4D) segmental cardiac function: displacement analysis of myocardial mechanical deformation (DIAMOND). Transgenic embryonic zebrafish were imaged in vivo using a light-sheet fluorescence microscopy system with 4D cardiac motion synchronization. Acquired 3D digital hearts were reconstructed at end-systole and end-diastole, and the ventricle was manually segmented into binary datasets. Then, the heart was reoriented and isotropically resampled along the true short axis, and the ventricle was evenly divided into eight portions (I-VIII) along the short axis. Due to the different resampling planes and matrices at end-systole and end-diastole, a transformation matrix was applied for image registration to restore the original spatial relationship between the resampled systolic and diastolic image matrices. After image registration, the displacement vector of each segment from end-systole to end-diastole was calculated based on the displacement of mass centroids in three dimensions (3D). DIAMOND shows that basal myocardial segments adjacent to the atrioventricular canal undergo the highest mechanical deformation and are the most susceptible to doxorubicin-induced cardiac injury. Overall, DIAMOND provides novel insights into segmental cardiac mechanics in zebrafish embryos beyond traditional ejection fraction (EF) under both physiological and pathological conditions.


Assuntos
Algoritmos , Embrião não Mamífero/fisiologia , Coração/embriologia , Coração/fisiologia , Processamento de Imagem Assistida por Computador , Peixe-Zebra/embriologia , Animais , Animais Geneticamente Modificados , Diástole/efeitos dos fármacos , Diástole/fisiologia , Doxorrubicina/farmacologia , Embrião não Mamífero/efeitos dos fármacos , Coração/efeitos dos fármacos , Testes de Função Cardíaca , Ventrículos do Coração/efeitos dos fármacos , Imageamento Tridimensional , Receptores Notch/metabolismo , Sístole/fisiologia
20.
Dev Cell ; 52(3): 350-363.e6, 2020 02 10.
Artigo em Inglês | MEDLINE | ID: mdl-31928974

RESUMO

The mammalian heart contains multiple cell types that appear progressively during embryonic development. Advance in determining cardiac lineage diversification has often been limited by the unreliability of genetic tracers. Here we combine clonal analysis, genetic lineage tracing, tissue transplantation, and mutant characterization to investigate the lineage relationships between epicardium, arterial mesothelial cells (AMCs), and the coronary vasculature. We report a contribution of the second heart field (SHF) to a vasculogenic niche composed of AMCs and sub-mesothelial cells at the base of the pulmonary artery. Sub-mesothelial cells from this niche differentiate into lymphatic endothelial cells and, in close association with AMC-derived cells, contribute to and are essential for the development of ventral cardiac lymphatics. In addition, regionalized epicardial/mesothelial retinoic acid signaling regulates lymphangiogenesis, contributing to the niche properties. These results uncover a SHF vasculogenic contribution to coronary lymphatic development through a local niche at the base of the great arteries.


Assuntos
Diferenciação Celular , Vasos Coronários/fisiologia , Endotélio Vascular/fisiologia , Coração/fisiologia , Linfangiogênese , Vasos Linfáticos/fisiologia , Pericárdio/fisiologia , Animais , Linhagem da Célula , Vasos Coronários/citologia , Endotélio Vascular/citologia , Epitélio/fisiologia , Feminino , Coração/embriologia , Vasos Linfáticos/citologia , Masculino , Camundongos , Pericárdio/citologia , Transdução de Sinais
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