Main description:

This volume provides the reader with detailed protocols that describe a variety of in vitro and in vivo techniques that study reductionist systems and human samples. The methods covered in this book explore a broad selection of topics such as functional aspects of alpha-synuclein biology, its lipid-interactions and misfolding, and functional deficits in cells and mice. Chapters also cover topics such as kinetic measurements of endocytosis and exocytosis in cultured neurons; electrochemiluminescence-based detection; yeast-based screens to target alpha-synuclein toxicity; mass-spectrometry of alpha-synuclein in rat cortical neurons; and expression and purification of untagged a-synuclein. 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 thorough, Alpha-Synuclein: Methods and Protocols is a valuable reference guide that aids researchers working on topics related to alpha-synuclein and its various aspects.

Contents:

Preface...Table of Contents...Contributing Authors... 1. Assays for Neuronal Defects Caused by Early Formation of -Synuclein Inclusions in Primary Cultured NeuronsLaura A. Volpicelli-Daley
2. Immunohistochemical Detection of Alpha-Synuclein in Unfixed Human Brain TissueJohn B. Sanderson
3. Kinetic Measurements of Endocytosis and Exocytosis in Cultured NeuronsKarina Vargas
4. Detection of Misfolded -Synuclein Aggregates in Cerebrospinal Fluid by the Protein Misfolding Cyclic Amplification PlatformLuis Concha-Marambio, Mohammad Shahnawaz, and Claudio Soto
5. Stereotaxic Targeting of Alpha-Synuclein Pathology in Mouse Brain using Pre-Formed FibrilsBin Zhang, Victoria Kehm, Ron Gathagan, Susan N. Leight, John Q. Trojanowski, Virginia M.-Y. Lee, and Kelvin C. Luk
6. Quantification of Alpha-Synuclein in Biological Fluids by Electrochemiluminescence-Based DetectionNiels Kruse and Brit Moellenhauer
7. Alpha-Synuclein Proximity Ligation Assay (AS-PLA) in Brain Sections to Probe for Alpha-Synuclein OligomersRosalind F. Roberts, Nora Bengoa-Vergniory, and Javier Alegre-Abarrategui
8. Studying -Synuclein Conformation by Intact-Cell CrosslinkingThibaut Imberdis, Saranna Fanning, Andrew Newman, Nagendran Ramalingam, and Ulf Dettmer
9. C. elegans as a Model for Synucleinopathies and Other Neurodegenerative Diseases: Tools and TechniquesMandy Koopman, Renee I. Seinstra, and Ellen. A. A. Nollen
10. Recombinant Expression and Purification of N-Acetylated Alpha-SynucleinTara A. Eastwood and Daniel P. Mulvihill
11. Circular Dichroism and Isothermal Titration Calorimetry to Study the Interaction of -Synuclein with MembranesMatteo Rovere
12. Yeast-Based Screens to Target Alpha-Synuclein ToxicityInes Caldeira Bras, Blagovesta Popova, Gerhard H. Braus, and Tiago F. Outeiro
13. Probing Structural Changes in Alpha Synuclein by Nuclear Magnetic Resonance SpectroscopyTapojyoti Das and David Eliezer
14. Measuring Mitochondrial Dysfunction Caused by Soluble -Synuclein OligomersEric S. Luth and Irina G. Stavrovskaya
15. Monitoring -Synuclein Proteotoxicity in Drosophila ModelsAron Szabo and George K. Tofaris
16. Detection of -Synuclein in Biological Samples using Mass SpectrometryAnn Brinkmalm, Annika OEhrfelt, Payel Bhattacharjee, and Henrik Zetterberg
17. Single-Molecule FRET Detection of Early-Stage Conformations in -Synuclein AggregationMahdi Muhammad Moosa, Josephine C. Ferreon, and Allan Chris M. Ferreon
18. In Situ Peroxidase Labelling and Mass-Spectrometry of Alpha-Synuclein in Rat Cortical NeuronsRicardo Sanz, Patrick Ovando-Roche, Namrata D. Udeshi, Steven A. Carr, Chee Yeun Chung, and Vikram Khurana
19. Probing Alpha-Synuclein Conformations by Electron Paramagnetic Resonance (EPR) SpectroscopyJulia Cattani, Theresa Braun, and Malte Drescher
20. Expression and Purification of Untagged -SynucleinAlex E. Powers and Dushyant S. Patel
21. Generation and Analysis of Viral Vector Mediated Rodent Models for Parkinson's DiseaseAkua A. Karikari, James B. Koprich, and Chi Wang Ip