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细胞功能测定

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Angiogenesis Assays in the Chick CAM

The growth of new blood vessels from pre-existing vascular elements, or angiogenesis, involves coordinated signals to the adhesion, migration, and survival machinery within the target endothelial cell. Agents that interfere with any of these processes may therefore influence ang ...

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Tumor Cell Invasion Assays

The ability of tumor cells to invade is one of the hallmarks of the metastatic phenotype. To elucidate the mechanisms by which tumor cells acquire an invasive phenotype, in vitro assays have been developed that mimic the in vivo process. The most commonly used in vitro invasion assay is a modified Boyd ...

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Analysis of Cell Migration Using Drosophila as a Model System

There are a number of reasons to use Drosophila as a model system to study cell migration. First and foremost is the availability of an arsenal of powerful genetic techniques that can be deployed, permitting the study of cell migration in vivo, in the context of the entire organism. This is especially imp ...

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Analysis of Cell Migration in Caenorhabditis elegans

This chapter is concerned with a method of analysis and quantification of cell migration defects in mutants of the nematode worm Caenorhabditis elegans. The method takes advantage of transgenic expression of the green fluorescent protein to visualize migrating cells. By following th ...

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Imaging Cell Movement During Dorsal Closure in Drosophila Embryos

During Drosophila embryogenesis, many cells and tissues undergo complex morphogenetic movements, such as ventral furrow formation, germ band extension and retraction, and dorsal closure. The best way to study and understand the cell behaviors during such tissue movements is to image ...

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Analysis of Cell Movements in Zebrafish Embryos: Morphometrics and Measuring Movement of Labeled Cell Populations In Vivo

Cell movements occur in all phases of animal life from embryogenesis, to maintaining adult organs, to comprising a critical component of pathology. During gastrulation, cells demonstrate a repertoire of morphogenetic movements coordinated with fate inductions to sculpt the embr ...

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The Xenopus Embryo as a Model System for Studies of Cell Migration

In this chapter, we describe procedures for the microsurgical removal of cells and tissues from early-stage embryos of the amphibian Xenopus laevis. Using simple culture conditions and artificial substrates, these preparations undergo a variety of quantifiable cellular behavi ...

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Neural Crest Migration Methods in the Chicken Embryo

Neural crest cells emerge from the neural tube early in development. They migrate extensively throughout the embryo and form most of the head and peripheral nervous system, giving rise to sensory and sympathetic ganglia, heart regions, adrenal cells, head bones, teeth, muscle cells, sensory ...

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In Vivo and In Vitro Models of Mammalian Angiogenesis

Angiogenesis is a complex process involving the organization of proliferating endothelial cells into new blood vessels. Both in vivo models and in vitro models are important for investigating angiogenesis and for defining the involvement of specific molecules. This chapter descr ...

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Measurement of Protein Tyrosine Phosphorylation in Cell Adhesion

This chapter describes biochemical, immunochemical, and microscopic approaches to measure protein tyrosine phosphorylation after cell adhesion. We have outlined detailed procedures to biochemically examine the phosphotyrosine content of cellular proteins by Weste ...

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Cell-Adhesion Assays: Fabrication of an E-Cadherin Substratum and Isolation of Lateral and Basal Membrane Patches

Cell adhesion between cells and with the extracellular matrix (ECM) results in dramatic changes in cell organization and, in particular, the cytoskeleton and plasma membrane domains involved in adhesion. However, current methods to analyze these changes are limited because of the sma ...

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Use of Multiphoton Imaging for Studying Cell Migration in the Mouse

We describe a method for studying cell motility in the living mouse using multiphoton microscopy. The procedure consists of mouse anesthesia, labeling of target cells with enhanced green fluorescent protein by infection with recombinant adenovirus, implantation of beads carryi ...

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Application of Microscope-Based FRET to Study Molecular Interactions in Focal Adhesions of Live Cells

This chapter describes the use of microscope-based fluorescence resonance energy transfer to follow dynamic interaction of molecules localized at focal adhesions. We first outline the significance of studying dynamic interactions in focal adhesions of living cells, and second ...

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Using Microfluidic Channel Networks to Generate Gradients for Studying Cell Migration

In this chapter, we will discuss a method for the generation of gradients that can be quantitatively used for studying directional cell migration. Microfluidic networks, which serially split and remix small volumes of solutions under laminar flow conditions to generate a series of micro ...

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Assays of Adrenergic Receptors: Including Lipolysis and Binding Measurements

Adipocytes express the three major types of adrenoceptors (ARs), α1-, α2-, and β-ARs, each of them being further divided into three subtypes. With the exception of α1A-, α1B-, and α1D-ARs, which have not been described to directly influence cAMP production in fat cells, all the other AR subtypes are known to ...

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Assay of Membrane Transport of Long-Chain Fatty Acids by Adipocytes

As a result of their poor aqueous solubility, Fatty acids (FAs) are quantitatively bound to plasma albumin in the circulation and to cytosolic FA-binding proteins inside the cell. The presence of FA-binding proteins on both sides of the plasma membrane complicates interpretation of FA upta ...

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Transfection of Adipocytes and Preparation of Nuclear Extracts

The uniqueness of the adipose cell type must be considered before choosing a procedure for gene transfer into mature adipocytes. Because adipocytes isolated from adipose tissue (AT) are filled with so many lipids that cells cannot be plated on a culture support, because of their low density, and b ...

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Assays of Glucose Entry, Glucose Transporter Amount, and Translocation

Glucose enters the cell by a carrier-mediated, facilitated diffusion mechanism, which, in most tissues, exhibits no energy or counter-ion requirements. In adipose tissues and skeletal muscle, glucose entry is acutely regulated by insulin and other hormones (1,2). Indeed, in those tissu ...

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Methods to Study Phosphorylation and Activation of the Hormone-Sensitive Adipocyte Phosphodiesterase Type 3B in Rat Adipocytes

Cyclic nucleotide phosphodiesterases (PDEs) include a large group of structurally related enzymes that are responsible for the hydrolysis of cyclic adenosine monophosphate (cAMP) and cyclic guanosine monophosphate (cGMP). These enzymes belong to at least nine related gene fam ...

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Measurements of Glucose Conversion to its Metabolites

Glucose is the main substrate for most tissues of the body, and provides energy for cellular respiration and metabolic activities. The fate of glucose end-products depends, in part, on the physiological demands of the tissue in question. Similarly, the type and function of glucose transport ...

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