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This volume explores detailed methods to investigate various aspects of biology related to cell polarity, or asymmetry within a cell. Molecular, cellular, and tissue-level regulation and function as well as diseases caused by impairment of cell polarity are explored by these methods. Beginning with advanced imaging and biochemical methods, the book continues with planar cell polarity (PCP) signaling in morphogenesis in diverse developmental contexts, apicobasal (AB) cell polarity in development and diseases, as well as directional cell migration and biomechanics in cell polarity. The collection includes the usage of a wide variety of model systems and an extensive array of techniques, including genetic, imaging, biochemical, and biomechanical. Written for the highly successful Methods in Molecular Biology 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. Authoritative and comprehensive, Cell Polarity Signaling: Methods and Protocols aims to enable researchers to delve into the stimulating field of cell polarity and contribute to our understanding of how coordinated control of protein stability, trafficking, membrane retention, post-translational modification, and dynamic organization leads to active regulation of cell polarity.
Use of Fluorescence Recovery After Photobleaching (FRAP) to Measure In Vivo Dynamics of Cell Junction–Associated Polarity Proteins
FRET Imaging of Rho GTPase Activity with Red Fluorescent Protein-Based FRET Pairs
Live-Cell Total Internal Reflection Fluorescence (TIRF) Microscopy to Investigate Protein Internalization Dynamics
Single-Cell Single-Molecule Pull-Down (sc-SiMPull) for Detection of Protein Complexes from Embryonic Lysates
Biochemical Assays to Detect Activation of Small GTPases Rho, Rac, and Cdc42 during Morphogenesis
Analysis of Planar Cell Polarity Complexes by Proximity Biotinylation in Xenopus Embryos
Regulation of Cell Polarity by Posttranslational Protein Palmitoylation
Enrichment and Detection of Wnt Proteins from Cell Culture Media
Using Live Imaging to Examine Early Cardiac Development in Zebrafish
Imaging Planar Cell Polarity Proteins in Xenopus Neuroectoderm
In Ovo Gain- and Loss-of-Function Approaches to Study Gut Morphogenesis
Avian Embryos as a Model to Study Vascular Development
BAC Recombineering and Transgenesis to Study Cell Polarity and Polarized Tissue Morphogenesis in Mice
Two-Photon Cell and Tissue Level Laser Ablation Methods to Study Morphogenetic Biomechanics
Protocols for Investigating the Epithelial Properties of Cardiac Progenitor Cells in the Mouse Embryo
Methods to Investigate Cell Polarity of Inner Ear
Characterization of Axon Guidance Phenotypes in Wnt/PCP Mutant Mice
In Vitro Explant Assays and Cultures to Study PCP Signaling in Axon Guidance
Biochemical and Cellular Assays to Study Mechanisms of PCP Signaling in Axon Guidance
Dissection, Fixation, and Immunostaining of the Drosophila Midgut
In Vivo Analysis of Pathways Regulating Epithelial Polaritypolarity and Secretion Using Drosophila Salivary Glands
Imaging Epidermal Cell Rearrangement in the C. elegans Embryo
Methods for the Study of Apical Constrictionconstriction During Ascidian Gastrulation
Assays for Apical Constriction Using the Xenopus Model
The Use of Three-Dimensional Cell Culture to Study Apicobasal Polarization and Lumen Formation
Studying Cell Polarity Dynamics During Cancer Initiation Using Inducible 3D Organotypic Cultures
Under-Agarose Chemotaxis and Migration Assays for Dictyostelium
Mapping Asymmetry in Collective Cell Migration: Lessons from Border Cells in Drosophila Oogenesis
A Toolbox to Study Tissue Mechanics In Vivo and Ex Vivo
Quantitative Analysis of Directional Neural Crest Cell Migration