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Full Description
This volume provides the novice to expert tau scientist with detailed protocols to establish in vitro and in vivo techniques designed for understanding the physiopathology of tau in neurodegenerative diseases. The protocols in this book were intentionally selected to provide a basis for research laboratories focusing in tau neurodegenerative drug discovery, and are organized into four cohesive parts. Part One covers the procuration of recombinant tau protein, which is necessary to produce a particular isoform, induce hyperphosphorylation state, or capture oligomer conformation. Part Two describes techniques to obtain neurofibrillary tangles (insoluble tau) and soluble form of tau. Part Three focuses on conditions mimicking neurodegenerative scenarios in vitro such as oligomer formation, tau-RNA interaction, and seeding. Part Four provides protocols for robust proof of concept studies involving C. Elegan, drosophila, zebrafish, and rodent tau models. This book is part of the established Neuromethods book series, and each chapter includes short introduction to the topic, list of materials, laboratory protocol, recommendations, and troubleshooting section.
Cutting-edge and comprehensive, Translational Research Methods for Tauopathies is a valuable tool for researchers interested in learning more about this important field.
Contents
Production of Hyperphosphorylated Tau Using a Baculovirus-Insect Cell System.- Expression and Production of Tau in E. Coli Cells.- Preparation and Examination of Hyperphosphorylated Tau Produced by PIMAX System.- Structural and Functional Assays for the Characterization of Recombinant Hyperphosphorylated Tau Protein.- Evaluation of the Kinetics of Tau Fibril Formation.- Tracking Tau Oligomer Formation In Vitro.- Isolation and Characterization of Amyloid-b Plaque Cores and Neurofibrillary Tangles from Alzheimer's Disease Brain.- Isolation and Proteomic Analysis of Pathological Tau-Seeds.- Neurotoxic Tau Oligomers: Isolation, Preparation, and Characterization.- Capturing Tau and RNA Interactions.- Generation of Stable Tau Biosensor Cell Lines.- Cell-Based Tau Polymerization Assays.- Modelling Functions and Pathology of Tau Protein in C. elegans.- Tracking Tau Multimerization in the Adult Drosophila Brain.- Testing Small Molecules for Activity Against Neurological Phenotypes in a Zebrafish 4-Repeat Tauopathy Model.- Tau Transgenic Mice.
