Auditory and Vestibular Research. Methods and Protocols (Methods in Molecular Biology 493)

About this book

While early studies of the auditory/vestibular sciences provided insights into the anatomy and neurophysiology of these systems and produced a prosthetic cochlear implant, the rise of molecular biology now permits a clear examination of the genetic basis for various hearing and balance disorders. In Auditory and Vestibular Research: Methods and Protocols, specialists in the field describe current RNA, protein, and imaging protocols that have provided insights into genetic regulation as well as a greater understanding of the genes and pathogens involved in diseases of the ear. This overview utilizes both mammalian and non-mammalian animal models as well as techniques applicable to clinical studies in order to best provide an up-to-date perspective on basic research. Written in the highly successful Methods in Molecular Biology™ series format, chapters include brief introductions to their subjects, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and Notes sections, highlighting tips on troubleshooting and avoiding known pitfalls.

Comprehensive and cutting-edge, Auditory and Vestibular Research: Methods and Protocols is an ideal guide to the field and a useful tool for exploring genes and proteins in other systems as well, especially systems in which tissues are scarce and a comparative approach lends itself to discovering the underlying causes of human disorders.

Written for: Otolaryngologists, molecular biologists, pathologists

Keywords:

• Amino acid
• Balance disorders
• Hearing disorders
• Nucleic acid
• RT-PCR

Table of contents

Part 1: Nucleic Acid Protocols

• RNA Isolation from Xenopus Inner Ear Sensory Endorgans for Transcriptional Profiling and Molecular Cloning
  Casilda Trujillo-Provencio, TuShun R. Powers, David R. Sultemeier, and Elba E. Serrano
• Synthesis of Biotin-Labeled RNA for Gene Expression Measurements Using Oligonucleotide Arrays
  Ana E. Vázquez, Liping Nie, and Ebenezer N. Yamoah
• In situ Hybridization Approach to Study mRNA Expression and Distribution in Cochlear Frozen Sections
  Hakim Hiel
• Lineage Analysis of Inner Ear Cells Using Genomic Tags for Clonal Identification
  Takunori Satoh and Donna M. Fekete
• Genetic Fate Mapping Approaches: New Means to Explore the Embryonic Origins of the Cochlear Nucleus
  Jun Chul Kim and Susan M. Dymecki
• The Practical Use of Cre and loxP Technologies in Mouse Auditory Research
  Yiling Yu and Jian Zuo
• Helios® Gene Gun-Mediated Transfection of the Inner Ear Sensory Epithelium
  Inna A. Belyantseva
• Electroporation-Mediated Gene Transfer to the Developing Mouse Inner Ear
  John V. Brigande, Samuel P. Gubbels, David W. Woessner, Jonathan J. Jungwirth, and Catherine S. Bresee
• Isolation of Sphere-Forming Stem Cells from the Mouse Inner Ear
  Kazuo Oshima, Pascal Senn, and Stefan Heller
• Molecular Biology of Vestibular Schwannomas
  Long-Sheng Chang and D. Bradley Welling
• Multilocus Sequence Typing and Pulsed-Field Gel Electrophoresis of Otitis Media Causing Pathogens
  Jonathan C. Thomas and Melinda M. Pettigrew
• Fluorescence "In situ" Hybridization for the Detection of Biofilm in the Middle Ear and Upper Respiratory Tract Mucosa
  Laura Nistico, Armin Gieseke, Paul Stoodley, Luanne Hall-Stoodley, Joseph E. Kerschner, and Garth D. Ehrlich
• Positional Cloning of Deafness Genes
  Hannie Kremer and Frans P. M. Cremers

Part II: Amino Acid Protocols

• Twist-off Purification of Hair Bundles
  Jung-Bum Shin, James Pagana, and Peter G. Gillespie
• Yeast Two-Hybrid Screening to Test for Protein-Protein Interactions in the Auditory System
  Dhasakumar S. Navaratnam
• The Use of 2-D Gels to Identify Novel Protein-Protein Interactions in the Cochlea
  Thandavarayan Kathiresan, Margaret C. Harvey, and Bernd H. A. Sokolowski
• Identification of Functionally Important Residues/Domains in Membrane Proteins Using an Evolutionary Approach Coupled with Systematic Mutational Analysis
  Lavanya Rajagopalan, Fred A. Pereira, Olivier Lichtarge, and William E. Brownell
• In vivo Verification of Protein Interactions in the Inner Ear by Coimmunoprecipitation
  Margaret C. Harvey and Bernd H. A. Sokolowski
• Identification of Transcription Factor-DNA Interactions Using Chromatin Immunoprecipitation Assays
  Liping Nie, Ana E. Vazquez, and Ebenezer N. Yamoah
• Surface Plasmon Resonance (SPR) Analysis of Binding Interactions of Proteins in Inner-Ear Sensory Epithelia
  Dennis G. Drescher, Neeliyath A. Ramakrishnan, and Marian J. Drescher
• Multiplexed Isobaric Tagging Protocols for Quantitative Mass Spectrometry Approaches to Auditory Research
  Douglas E. Vetter, Johnvesly Basappa, and Sevin Turcan

Part III: Imaging Protocols

Fluorescence Microscopy Methods in the Study of Protein Structure and Function
Heather Jensen-Smith, Benjamin Currall, Danielle Rossino, LeAnn Tiede, Michael Nichols, and Richard Hallworth

Ion Imaging in Cochlear Hair Cells
Gregory I. Frolenkov

Atomic Force Microscopy in Studies of the Cochlea
Michio Murakoshi and Hiroshi Wada