This volume discusses the latest advancements and techniques used to study Dictyostelium discoideum. The chapters in this book cover topics such as cytokinesis, membrane function and damage, and nutrient/bacterial processing; autophagy activation and function; protein localization, interaction, and activation; cell migration and contact guidance for quantification of cytoskeletal dynamics; and global mutational screening, transcriptome analyses, and proteomics that apply to all stages of the Dictyostelium life cycle and that may generalize to human disease models. 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 tips on troubleshooting and avoiding known pitfalls.
Cutting-edge and comprehensive, Dictyostelium discoideum: Methods and Protocols is a valuable resource for all researchers who are interested in learning more about this important and developing field.
Discovery and Quantitative Dissection of Cytokinesis Mechanisms Using
Dictyostelium discoideum.- Superresolution Expansion Microscopy in
Dictyostelium Amoebae.- Inflicting, Monitoring, Visualizing, and Quantitating
Various Sterile Membrane Damage and the Repair Response in Dictyostelium
discoideum.- Assaying Lysosomal Enzyme Activity in Dictyostelium
discoideum.- Procedures to Measure Dictyostelium Phagocytosis and
Macropinocytosis.- Quantitative Analysis of the Inactivation Process of
Internalized Bacteria in Dictyostelium Cells.- Methods to Assess Autophagic
Activity in Dictyostelium.- Controlling the Subcellular Localization of
Signaling Proteins using Chemically-Induced Dimerization and
Optogenetics.- APEX2-Mediated Proximity Protein Labeling in
Dictyostelium.- GRminusRD, A Sensitive Assay to Detect Activation Processes
at the Plasma Membrane in Living Cells.- Analyzing the Micro-Topography
Guidance of Dictyostelium Cells using Microfabricated Structures.- Evaluating
the Activity of Ras/Rap GTPases in Dictyostelium.- Analysis of cGMP Signaling
in Dictyostelium.- Quantitative Monitoring of GPCR-Mediated Spatiotemporal
IP3 Dynamics Using Confocal Fluorescence Microscopy.- Mutation Screening of
Dictyostelium Restriction Enzyme-Mediated Integration (REMI)
Libraries.- Single Cell Transcriptome Analysis during Development in
Dictyostelium.- Proteomic Analysis of Dictyostelium discoideum by Mass
Spectrometry.
