Tau Protein, 1st ed. 2017
Methods and Protocols

Methods in Molecular Biology Series, Vol. 1523

Coordinator: Smet-Nocca Caroline

Language: Anglais

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This detailed volume gathers basic and advanced methods and protocols from in vitro assays and in vivo models to address the molecular and functional aspects of tau physiopathology. Divided into five parts that illustrate the underlying molecular mechanisms of Tau functions and dysfunctions in Alzheimer?s disease and other dementia referred to as tauopathies, this volume discusses timely topics such as: conformational studies of native tau protein and investigation of its physiological function in microtubules binding and tubulin polymerization; in vitro methods of formation and detection of tau oligomers and PHF-like fibrils; characterization and in vitro introduction of post-translational modifications on tau proteins for further functional studies; analytical tools for the detection of tau proteins, their modifications and cellular interactions, and MAPT gene mutations in various biological samples; and cellular and in vivo models for the investigations of tau physiopathology. 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.

Practical and cutting-edge, Tau Protein: Methods and Protocols is a valuable resource that addresses the most recent aspects of Alzheimer?s disease research related to Tau protein and state-of-the-art experimental techniques. It targets a broad scientific audience including molecular biologists and biochemists, as well as all researchers interested in exploring the functions of tau proteins.

Table of Contents


Part I Native Tau Protein: Conformational Studies and Tubulin Interactions

1.  Conformational Dynamics of Intracellular Tau Protein Revealed by CD and SAXS
Nalini Vijay Gorantla, Alexander V. Shkumatov, and Subashchandrabose Chinnathambi

2. Global Conformation of Tau Protein Mapped by Raman Spectroscopy
Nalini Vijay Gorantla, Puneet Khandelwal, Pankaj Poddar, and Subashchandrabose Chinnathambi

3.  Molecular Dynamics Simulation of Tau Peptides for the Investigation of Conformational Changes Induced by Specific Phosphorylation Patterns
Neha S.Gandhi, Predrag Kukic, Guy Lippens, and Ricardo L. Mancera

4. Tau Interaction with Tubulin and Microtubules: From Purified Proteins to Cells
Tiphany De Bessa, Gilles Breuzard, Diane Allegro, François Devred, Vincent Peyrot, and 
Pascale Barbier

Part II In Vitro Self-Association of Tau Protein: Oligomer and Fibril Formation

5. X-Ray Structural Study of Amyloid-Like Fibrils of Tau Peptides Bound to Small-Molecule Ligands
Einav Tayeb-Fligelman and Meytal Landau

6. Detection and Quantification Methods for Fibrillar Products of In Vitro Tau Aggregation Assays
Niki Nanavaty, Lauren Lin, Samantha H. Hinckley, and Jeff Kuret

7. Fourier Transform Infrared (FTIR) Spectroscopy, Ultraviolet Resonance Raman (UVRR) Spectroscopy and Atomic Force Microscopy (AFM) for Study of the Kinetics of Formation and Structural Characterization of Tau Fibrils
Gayathri Ramachandran 

8. Assays for the Screening and Characterization of Tau Aggregation Inhibitors
Janet E. Rickard, David Horsley, Claude M. Wischik, and Charles R. Harrington

9. Tau Oligomers as Pathogenic Seeds: Preparation and Propagation In Vitro and In Vivo
Julia E. Gerson, Urmi Sengupta, and Rakez Kayed
Part III Tau Post-Translational Modifications

10. Mass Spectrometry Analysis of Lysine Post-Translational Modifications of Tau Protein from Alzheimer’s Disease Brain
Stefani N. Thomas and Austin J. Yang

11. The Study of Posttranslational Modifications of Tau Protein by Nuclear Magnetic Resonance Spectroscopy: Phosphorylation of Tau Protein by ERK2 Recombinant Kinase and Rat Brain Extract, and Acetylation by Recombinant Creb-Binding Protein
Haoling Qi, Clément Despres, Sudhakaran Prabakaran, François-Xavier Cantrelle, Béatrice 
Chambraud, Jeremy Gunawardena, Guy Lippens, Caroline Smet-Nocca, and Isabelle Landrieu

12. Tag-Free Semi-Synthesis of Tau Protein
Oliver Reimann, Caroline Smet-Nocca, and Christian P. R. Hackenberger

13. Production of O-GlcNAc Modified Recombinant Tau in E.coli and Detection of Ser400 O-GlcNAc Tau In Vivo
Scott A. Yuzwa and David J. Vocadlo

Part IV Detecting Tau Proteins, Their Modifications, Mutations and Interacting Partners

14. Two-Dimensional Electrophoresis Protocols to Analyze the Microtubule-Associated Tau Proteins from Several Biological Sources
Nicolas Sergeant, Francisco-Jose Fernandez-Gomez, Helene Obriot, Sabiha Eddarkaoui,
Valérie Buée-Scherrer, and Luc Buée 

15. A Simple Method to Avoid Non-Specific Signal When Using Monoclonal Anti-Tau Antibodies in Western Blotting of Mouse Brain Proteins
Franck R. Petry, Samantha B. Nicholls, Sébastien S. Hébert, and Emmanuel Planel

16. Flow Cytometry Analysis and Quantitative Characterization of Tau in Synaptosomes from Alzheimer’s Disease Brains
Karen Hoppens Gylys and Tina Bilousova

17. In Vivo Microdialysis of Brain Interstitial Fluid for the Determination of Extracellular Tau Levels
Kaoru Yamada

18. Proximity Ligation Assay: A Tool to Study Endogenous Interactions between Tau and its Neuronal Partners
Alexis Bretteville, Florie Demiautte, and Julien Chapuis

19. Finding MAPT Mutations in Frontotemporal Dementia and Other Tauopathies
Carol Dobson-Stone and John B. J. Kwok

Part V Cellular and In Vivo Models of Tau Physiopathology

20. Tracking Tau in Neurons: How to Grow, Fix and Stain Primary Neurons for the Investigation of Tau in All Developmental Stages
Hans Zempel and Eva-Maria Mandelkow

21. Tracking Tau in Neurons: How to Transfect and Track Exogenous Tau into Primary Neurons
Hans Zempel, Julia Lüdtke, and Eva-Maria Mandelkow

22. Image-Based Analysis of Intracellular Tau Aggregation by Using Tau-BiFC Cell Model
Sungsu Lim, Dohee Kim, Dong Jin Kim, and Yun Kyung Kim

23. FRET and Flow Cytometry Assays to Measure Proteopathic Seeding Activity in Biological Samples
Jennifer L. Furman and Marc I. Diamond

24. In Vivo Imaging of Tau Aggregates in the Mouse Retina
Christian Schön and Jochen Herms

25. In Vivo Hyperthermic Stress Model: An Easy Tool to Study the Effects of Oxidative Stress on Neuronal Tau Functionality in Mouse Brain
Alban Chauderlier, Lucie Delattre, Luc Buée, and Marie-Christine Galas

26. Identification of Tau Toxicity Modifiers in the Drosophila Eye
Pierre Dourlen

27. Regulation of Neurotrophic Factors during Pathogenic Tau-Aggregation: A Detailed Protocol for Double-Labeling mRNA by In Situ Hybridization and Protein Epitopes by Immunohistochemistry
Katharina Schindowski

28. Pin1 Knockout Mice: A Model for the Study of Tau Pathology in Alzheimer's Disease
Asami Kondo, Onder Albayram, Xiao Zhen Zhou, and Kun Ping Lu

Includes cutting-edge methods and protocols

Provides step-by-step detail essential for reproducible results

Contains key notes and implementation advice from the experts