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Main description:
Since the cloning of the cystic fibrosis transmembrane conductance re- lator (CFTR) nearly a decade ago, cystic fibrosis (CF) research has witnessed a dramatic expansion into new scientific areas. Basic researchers, clinicians, and patients increasingly rely on fundamental techniques of genetics, molecular biology, electrophysiology, biochemistry, cell biology, microbiology, and immunology to understand the molecular basis of this complex disease. Research into the pathophysiology of CF has established numerous paradigms of ion channel dysfunction that extend from inflammation and infection in the airways of patients to basic mechanisms of protein processing and regulation in intracellular components. With these rapid advances has come an increasing need for research scientists to understand and utilize a growing array of basic laboratory tools. This volume of Methods in Molecular Medicine, Cystic Fibrosis Methods and Protocols satisfies that need by providing detailed protocols for the laboratory techniques used throughout CF research. From electrophysiology and cell biology, to animal models and gene therapy, the comprehensive set of methods covered here provide step-by-step instructions needed for investigators to incorporate new approaches into their research programs. Contributions have been chosen to reflect the rich diversity of techniques and to provide a cohesive framework for understanding challenges that are currently at the forefront of CF research. It is hoped that this volume will serve as a valuable reference that will not only foster interdisciplinary investigations into current problems encountered in CF, but also facilitate the translation of new scientific discoveries into clinical solutions.
Contents:
Part I. Genetics of Cystic Fibrosis
CFTR Mutation Detection by Multiplex Heteroduplex (mHET) Analysis on MDE Gel
Julian Zielenski, Isabel Aznarez, Tuncer Onay, John Tzounzouris, Danuta Markiewicz, and Lap-Chee Tsui
cDNA Microarrays for Pharmacogenomic Analysis of Cystic Fibrosis, Meera Srivastava
Ofer Eidelman, and Harvey B. Pollard
Natural Animal Models of Human Genetic Diseases
Jeffrey J. Wine, Michael Dean, and Damjan Glavac
Part II. CFTR Structure and Function: Structure, Gating, and Regulation
Electrophysiological Approach to Studying CFTR
Horst Fischer
Quantitative Analysis of ATP-Dependent Gating of CFTR
Allan Powe, Zhen Zhou, Tzyh-Chang Hwang, and Georg Nagel
CFTR Regulation by Phosphorylation
Tang Zhu, Deborah A. R. Hinkson, David Dahan, Alexandra Evagelidis, and John W. Hanrahan
Transepithelial Measurements of Bicarbonate Secretion in Calu-3 Cells
Robert J. Bridges
Transepithelial Impedance Analysis of Chloride Secretion
Ashvani K. Singh, Sangeeta Singh, Daniel C. Devor, Raymond A. Frizzell, Willy van Driessche, and Robert J. Bridges
Studies of the Molecular Basis for Cystic Fibrosis Using Purified Reconstituted CFTR Protein
Ilana Kogan, Mohabir Ramjeesingh, Canhui Li, and Christine E. Bear
Probing CFTR Channel Structure and Function Using the Substituted-Cysteine-Accessibility Method
Myles H. Akabas
Methods for the Study of Intermolecular and Intramolecular Interactions Regulating CFTR Function
Anjaparavanda P. Naren
Fluorescent Indicator Methods to Assay Functional CFTR Expression in Cells
Alan S. Verkman and Sujatha Jayaraman
Expression, Folding, and Degradation
Immunolocalization of CFTR in Intact Tissue and Cultured Cells
Christopher R. Marino
Analysis of CFTR Trafficking and Polarization Using Green Fluorescent Protein and Confocal Microscopy
Bryan D. Moyer and Bruce A. Stanton
CFTR Folding and Maturation in Cells
Mohamed Benharouga, Manu Sharma, and Gergely L. Lukacs
Isolation of CFTR: Chaperone Complexes by Co-Immunoprecipitation
Geoffrey C. Meacham and Douglas M. Cyr
CFTR Expression and ER-Associated Degradation in Yeast
Yimao Zhang, Susan Michaelis, and Jeffrey L. Brodsky
Manipulating the Folding Pathway of -F508 CFTR Using Chemical Chaperones
Marybeth Howard and William J. Welch
CFTR Degradation and Aggregation
Michael J. Corboy, Philip J. Thomas, and W. Christian Wigley
In Vitro Reconstitution of CFTR Biogenesis and Degradation
Jon Oberdorf and William R. Skach
In Vitro CFTR Folding Assays
Rhesa D. Stidham, W. Christian Wigley, and Philip J. Thomas
Analysis of CFTR Endocytosis by Cell Surface Biotinylation
Kelly Weixel and Neil A. Bradbury
Regulatory Complexes
CFTR Regulation of ENaC
Scott H. Donaldson, Elaine G. Poligone, and M. Jackson Stutts
Yeast Two-Hybrid Identification and Analysis of Protein Interactions with CFTR
Viswanathan Raghuram, Kenneth R. Hallows, and J. Kevin Foskett
Biochemical Assays for Studying Indirect Interactions Between CFTR and the Cytoskeleton
Peter J. Mohler, Patricia L. Kultgen, M. Jackson Stutts, and Sharon L. Milgram
CFTR-Associated ATP Transport and Release
Marie E. Egan
Part III. Pathophysiology of Cystic Fibrosis
Inflammatory Mediators in CF Patients,
Jay B. Hilliard, Michael W. Konstan, and Pamela B. Davis
Bacterial Colonization and Infection in the CF Lung
Scott D. Sagel, Elaine B. Dowell, and Frank J. Accurso
Antimicrobial Peptides and Proteins in the CF Airway,
Alexander M. Cole and Tomas Ganz
Bacterial-Epithelial Interactions
Ruth Bryan and Alice Prince
PRODUCT DETAILS
Publisher: Springer (Humana Press Inc.)
Publication date: November, 2011
Pages: 648
Weight: 979g
Availability: Available
Subcategories: Diseases and Disorders, General Issues, Respiratory Medicine
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