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. Preface… Table of Contents… Contributing Authors… 1. Assays for Neuronal Defects Caused by Early Formation of a-Synuclein Inclusions in Primary Cultured Neurons Laura A. Volpicelli-Daley 2. Immunohistochemical Detection of Alpha-Synuclein in Unfixed Human Brain Tissue John B. Sanderson 3. Kinetic Measurements of Endocytosis and Exocytosis in Cultured Neurons Karina Vargas 4. Detection of Misfolded a-Synuclein Aggregates in Cerebrospinal Fluid by the Protein Misfolding Cyclic Amplification Platform Luis Concha-Marambio, Mohammad Shahnawaz, and Claudio Soto 5. Stereotaxic Targeting of Alpha-Synuclein Pathology in Mouse Brain using Pre-Formed Fibrils Bin 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 Detection Niels Kruse and Brit Mllenhauer 7. Alpha-Synuclein Proximity Ligation Assay (AS-PLA) in Brain Sections to Probe for Alpha-Synuclein Oligomers Rosalind F. Roberts, Nora Bengoa-Vergniory, and Javier Alegre-Abarrategui 8. Studying a-Synuclein Conformation by Intact-Cell Crosslinking Thibaut Imberdis, Saranna Fanning, Andrew Newman, Nagendran Ramalingam, and Ulf Dettmer 9. C. elegans as a Model for Synucleinopathies and Other Neurodegenerative Diseases: Tools and Techniques Mandy Koopman, Rene I. Seinstra, and Ellen. A. A. Nollen 10. Recombinant Expression and Purification of N-Acetylated Alpha-Synuclein Tara A. Eastwood and Daniel P. Mulvihill 11. Circular Dichroism and Isothermal Titration Calorimetry to Study the Interaction of a-Synuclein with Membranes Matteo Rovere 12. Yeast-Based Screens to Target Alpha-Synuclein Toxicity Ins Caldeira Brs, Blagovesta Popova, Gerhard H. Braus, and Tiago F. Outeiro 13. Probing Structural Changes in Alpha Synuclein by Nuclear Magnetic Resonance Spectroscopy Tapojyoti Das and David Eliezer 14. Measuring Mitochondrial Dysfunction Caused by Soluble a-Synuclein Oligomers Eric S. Luth and Irina G. Stavrovskaya 15. Monitoring a-Synuclein Proteotoxicity in Drosophila Models Aron Szabo and George K. Tofaris 16. Detection of a-Synuclein in Biological Samples using Mass Spectrometry Ann Brinkmalm, Annika hrfelt, Payel Bhattacharjee, and Henrik Zetterberg 17. Single-Molecule FRET Detection of Early-Stage Conformations in a-Synuclein Aggregation Mahdi Muhammad Moosa, Josephine C. Ferreon, and Allan Chris M. Ferreon 18. In Situ Peroxidase Labelling and Mass-Spectrometry of Alpha-Synuclein in Rat Cortical Neurons Ricardo 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) Spectroscopy Julia Cattani, Theresa Braun, and Malte Drescher 20. Expression and Purification of Untagged a-Synuclein Alex E. Powers and Dushyant S. Patel 21. Generation and Analysis of Viral Vector Mediated Rodent Models for Parkinson’s Disease Akua A. Karikari, James B. Koprich, and Chi Wang Ip
