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Isolation and Characterization of Vascular Endothelial Cells from Murine Heart and Lung

The formation of blood vessel networks is a fundamental event in vertebrate embryo development. Angiogenesis and vasculogenesis are the essential processes in vascular formation. Endothelial cells play a key role during angiogenesis and vasculogenesis, and cultured vascular ...

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Indirect Immunostaining on Mouse Embryonic Heart for the Detection of Proliferated Cardiomyocyte

The heart is the first organ to form and become functional in a developing embryo and its proper function is critical for most, if not all, subsequent stages of an animal’s development. The formation of the heart relies heavily upon the rapid proliferation of cardiomyocytes at a specific stage during ...

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Analysis of Neural Crest Cell Fate During Cardiovascular Development Using Cre-Activated lacZ/-Galactosidase Staining

It is important to identify the mechanisms regulating cardiovascular development. However, complex genetic tools are often required, including transgenic animals that express the lacZ transgene encoding the β-galactosidase enzyme under the control of a specific promoter or fo ...

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Quantification of Myocyte Chemotaxis: A Role for FAK in Regulating Directional Motility

Formation of a fully functional four-chambered heart involves an intricate and complex series of events that includes precise spatial–temporal regulation of cell specification, proliferation, and migration. The formation of the ventricular septum during mid-gestation ens ...

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Analysis of the Endocardial-to-Mesenchymal Transformation of Heart Valve Development by Collagen Gel Culture Assay

Malformations of heart valves are one of the most common serious congenital defects. Heart valves are developed from endocardial cushions of the heart. The endocardial cushion in early heart development consists of two cell layers: an outer myocardial cell layer and an inner endocardial c ...

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Miniaturized Assays of Angiogenesis In Vitro

Assays of angiogenesis in vitro provide insights into vascular development and are useful in studies of agents that modulate blood vessel formation. This chapter describes techniques to induce angiogenesis-like sprouting from aortic and microvascular explants cultured in 3-d ...

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Visualization of Mouse Embryo Angiogenesis by Fluorescence-Based Staining

The establishment of a blood vessel network is fundamental to embryonic development and plays a critical role in many diseases including coronary heart disease and cancer. Vascular endothelial cells are central players in blood vessel formation and line the inside of the entire blood ves ...

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Whole-Mount Confocal Microscopy for Vascular Branching Morphogenesis

We introduce a whole-mount immunohistochemistry method for analyzing intricate vascular network formation in mouse embryonic tissues. Laser scanning confocal microscopy with multiple labeling allows for robust imaging of blood and lymphatic vessel branching morphogen ...

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Visualizing Vascular Networks in Zebrafish: An Introduction to Microangiography

Visualizing the circulatory pattern in developing embryos becomes an essential technique for the field of cardiovascular biology. In the zebrafish model system, there are currently several techniques available to visualize the circulatory pattern. Microangiography is a sim ...

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Chicken Chorioallantoic Membrane Angiogenesis Model

The chick embryo chorioallantoic membrane (CAM) is an extraembryonic membrane which serves as a gas exchange surface and its function is supported by a dense capillary network. Because of its extensive vascularization and easy accessibility, the CAM has been broadly used to study the morp ...

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Morpholino Injection in Xenopus

The study of gene function in developmental biology has been significantly furthered by advances in antisense technology made in the early 2000s. This was achieved, in particular, by the introduction of morpholino (MO) oligonucleotides. The introduction of antisense MO oligonucle ...

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Analysis of the Patterning of Cardiac Outflow Tract and Great Arteries with Angiography and Vascular Casting

Formation of the cardiac outflow tract and great arteries involves complex morphogenetic processes, whose abnormities result in several clinically important diseases. Studies of these developmental processes are therefore important for understanding congenital vasc ...

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Quantifying Cardiac Functions in Embryonic and Adult Zebrafish

Zebrafish embryos have been extensively used to study heart development and cardiac function, mainly due to the unique embryology and genetics of this model organism. Since most human heart disease occurs during adulthood, adult zebrafish models of heart disease are being created to dis ...

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Use of Whole Embryo Culture for Studying Heart Development

Congenital heart defects occur in approximately 1% of newborns and are a major cause of morbidity and mortality in infants and children. Many adult cardiac diseases also have developmental basis, such as heart valve malformations, among others. Therefore, dissecting the developmental ...

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Adenovirus-Mediated Gene Transfection in the Isolated Lymphatic Vessels

The authors describe technical details of experimental protocol of gene transfection in isolated rat mesenteric lymphatic vessels (MLVs). Authors also refer to the recent publication in Microcirculation, which provides wide set of experimental evidences obtained from confo ...

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Isolation of Cardiac Myocytes and Fibroblasts from Neonatal Rat Pups

Neonatal rat ventricular myocytes (NRVM) and fibroblasts (FBs) serve as in vitro models for studying fundamental mechanisms underlying cardiac pathologies, as well as identifying potential therapeutic targets. Both cell types are relatively easy to culture as monolayers and can be ...

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Application of Atomic Force Microscopy Measurements on Cardiovascular Cells

The atomic force microscope (AFM) is a state-of-the-art tool that can analyze and characterize samples on a scale from angstroms to 100 μm by physical interaction between AFM cantilever tip and sample surface. AFM imaging has been used incrementally over last decade in living cells in cardiova ...

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The Application of Genome-Wide RNAi Screens in Exploring Varieties of Signaling Transduction Pathways

Cardiovascular development is a precisely coordinated process at multilevels. It involves cross-talking among numerous signaling transduction pathways to ensure proper cell polarity, migration, proliferation, differentiation, and programmed death. Here, genome-w ...

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In Utero Assessment of Cardiovascular Function in the Embryonic Mouse Heart Using High-Resolution Ultrasound Biomicroscopy

Murine models are currently the preferred approach for studying the molecular mechanisms of cardiac dysfunction resulting from changes in gene expression. Transgenic and gene-targeting methods can be used to generate mice with altered cardiac size and function, and as a result, in vivo t ...

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Isolation and Preparation of RNA from Rat Blood and Lymphatic Microvessels for Use in Microarray Analysis

DNA microarray methodologies have proven to be an indispensable tool for genome-wide transcriptional profiling of organs, tissues, and cells. Here, we present a protocol for the optimized isolation and preparation of RNA from rat microvessels (including arteries, veins, and lympha ...

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