Dictyostelium discoideum Methods and Protocols

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.