Chemotaxis

Comprehensive and cutting edge, this practical guide covers diverse methodologies that are currently propelling chemotaxis research forward. Chapters include lists of necessary materials, step-by-step protocols, and notes on troubleshooting and pitfalls.

Fundamental to the development and vital functions of organisms, the migration of motile cells due to the detection of shallow gradients of specific chemical signals in their environments, or chemotaxis, can be clearly seen as a major force in cell biology. In Chemotaxis: Methods and Protocols , expert researchers in the field provide state-of-the-art methods for investigating cell migration behaviors, studying molecular components involved in detecting extracellular signals and directing cell movement, visualizing spatiotemporal dynamics of the components in signaling networks of chemotaxis in real time, and constructing quantitative models that simulate chemoattractant-induced cell responses. Written in the highly successful Methods in Molecular Biology ™ series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and notes on troubleshooting and avoiding known pitfalls.

Comprehensive and cutting-edge, Chemotaxis: Methods and Protocols serves scientists with practical guidance on the diverse methodologies that are currently propelling chemotaxis research forward in order to further our understanding of this vital biological system.

Describes a breadth of uniquely suited model systems for analysis of chemotaxis, including bacteria, Dictyostelium, yeast, Drosophila, zebrafish, neurons, and immune and tumor cells Examines phenomena related to chemotaxis, including chemotropism, phototaxis, and electrotaxis Details the application of cutting-edge technologies for stimulating cells with chemoattractants and analyzing their resulting cellular behaviors Covers a variety of state-of-the-art microscopic methods for visualizing the migration of cells in tissues and the spatiotemporal dynamics of chemotaxis signaling molecules Provides step-by-step functional assays for a variety of components involved in chemotaxis, including cell-surface receptors, downstream effectors, and cytoskeletal components Explores the quantitative description of chemoattractant gradients and computational approaches for modeling the signaling networks that underlie gradient sensing Includes supplementary material: sn.pub/extras

Inhalt
Microfluidic Techniques for the Analysis of Bacterial Chemotaxis.- Prokaryotic Phototaxis.- Photoorientation in Photosynthetic Flagellates.- Dictyostelium Slug Phototaxis.- Electrotaxis and Wound Healing: Experimental Methods to Study Electric Fields as a Directional Signal for Cell Migration.- Chemotropism During Yeast Mating.- Group Migration and Signal Relay in Dictyostelium.- Quantitative Analysis of Distal Tip Cell Migration in C. elegans.- Inflammation and Wound Healing in Drosophila.- Neutrophil Motility In Vivo Using Zebrafish.- Chemotaxis in Neutrophil-Like HL-60 Cells.- Chemokine Receptor Dimerization and Chemotaxis.- Intravital Two-Photon Imaging of Adoptively Transferred B Lymphocytes in Inguinal Lymph Nodes.- Breast Cancer Cell Movement: Imaging Invadopodia by TIRF and IRM Microscopy.- In Vivo Assay for Tumor Cell Invasion.- Quantitative Studies of Neuronal Chemotaxis in 3D.- Assays for Chemotaxis and Chemoattractant-Stimulated TorC2 Activation and PKB Substrate Phosphorylation in Dictyostelium.- Biochemical Responses to Chemoattractants in Dictyostelium: Ligand-Receptor Interactions and Downstream Kinase Activation.- Quantifying In Vivo Phosphoinositide Turnover in Chemotactically Competent Dictyostelium Cells.- In Vivo Measurements of Cytosolic Calcium in Dictyostelium discoideum.- Chemokine Receptor Signaling and HIV Infection.- Spatiotemporal Stimulation of Single Cells Using Flow Photolysis.- Spatiotemporal Regulation of Ras-GTPases During Chemotaxis.- FRAP Analysis of Chemosensory Components of Dictyostelium.- Monitoring Dynamic GPCR Signaling Events Using Fluorescence Microscopy, FRET Imaging, and Single-Molecule Imaging.- Imaging Actin Cytoskeleton Dynamics in Dictyostelium Chemotaxis.- Analysis of Actin Assembly by In Vitro TIRF Microscopy.- Single-Molecule Imaging Techniques to Visualize Chemotactic Signaling Events on the Membrane of Living Dictyostelium Cells.- Imaging B-Cell Receptor Signaling by Single-Molecule Techniques.- Light Microscopy to Image and Quantify Cell Movement.- Mathematics of Experimentally Generated Chemoattractant Gradients.- Modeling Spatial and Temporal Dynamics of Chemotactic Networks.- Computational Modeling of Signaling Networks for Eukaryotic Chemosensing.