Retinoid Protocols

ISBN-13: 9780896034389 / Angielski / Twarda / 1998 / 434 str.

Christopher Redfern

Opis

Interest in retinoic acid, the main biologically active derivative of vi- min A or retinol, increased dramatically between 1989 and 1993, following the cloning of nuclear receptors or RARs reported in 1987 (Fig. 1). Important discoveries since then have shown how RARs work as all-trans retinoic ac- dependent heterodimers with related nuclear receptors for 9-cis retinoic acid called RXRs. This has stimulated the development of synthetic analogs s- cific for each type of receptor, and opens the way to develop new methods for regulating pharmacologically the activity ofretinoic acid-dependent pathways of gene activation. The potential for the development of new drugs by the pharmaceutical industry is now a maj or factor driving forward our understa- ing of vitamin A-regulated pathways in animal development and homeostasis. However, apart from the real potential ofretinoid analogs as novel pharma- logical agents, there remains the considerable intellectual challenge of und- standing the way in which vitamin A and its derivatives function in cell development and differentiation. Retinoid Protocols is an attempt to bring together various methodologies that will be vital for rising to this challenge in the future. Retinoid molecular biology has few methods of its own, but is reliant on standard molecular biology methods applied to this particular research area.

Spis treści

Part I. Handling and Analysis of Retinoids Properties of Retinoids:Structure, Handling, and Preparation Arun B. Barua and Harold C. Furr Quantitative Analyses of Naturally Occurring Retinoids Joseph L. Napoli and Ronald L. Horst Detection and Measurement of Retinoic Acid Production by Isolated Tissues Using Retinoic Acid-Sensitive Reporter Cell Lines Michael Wagner Part II. Retinoid-Binding Proteins Immunohistochemistry for CRBPs and CRABPs Ulf Eriksson and Anne-Lee Gustafsson Whole-Mount In Situ Hybridization of Mouse Embryos Exposed to Teratogenic Levels of Retinoic Acid Sally D. Lyn Reverse Transcriptase-Polymerase Chain Reaction (RT-PCR) for Cellular Retinoid-Binding Proteins Andrew D. Loughney and Christopher P. F. Redfern Methods for Producing Recombinant Human Cellular Retinaldehyde-Binding Protein John W. Crabb, Yang Chen, Steve Goldflam, Karen West, and James Kapron Expression and Purification of CRABPs from E. coli Joseph L. Napoli, Zuzana Sperkova, Marek Sperka, and Paul D. Fiorella Purification and Fluorescent Titration of Cellular Retinol-Binding Protein Giorgio Malpeli, Claudia Folli, Davide Cavazzini, Giovanni Sartori, and Rodolfo Berni Fluorometric Titration of the CRABPs Andrew W. Norris and Ellen Li Expression and Mutagenesis of Retinol-Binding Protein Manickavasagam Sundaram, Asipu Sivaprasadarao, and John B. C. Findlay Interactions of Retinol-Binding Protein with Transthyretin and Its Receptor Asipu Sivaprasadarao, Manickavasagam Sundaram, and John B. C. Findlay Detection of Conformational Changes in Cellular Retinoid-Binding Proteins by Limited Proteolysis Robert S. Jamison, Marcia E. Newcomer, and David E. Ong Measurement of Rates of Dissociation of Retinoids from the Interphotoreceptor Retinoid-Binding Protein Noa Noy Use of Antisense Oligonucleotides to Study the Role of CRABPs in Retinoic Acid-Induced Gene Expression Paul Nugent and Robert M. Greene Part III. Nuclear Retinoid Receptors Preparation of Polyclonal Antibodies to Retinoid Receptors Jacqueline A. Dyck and Vickie J. LaMorte Detection of RARs and RXRs in Cells and Tissues Using Specific Ligand-Binding Assays and Ligand-Binding Immunoprecipitation Techniques Elizabeth A. Allegretto Nonisotopic In Situ Hybridization for the Detection of Nuclear Retinoid Receptor Transcripts in Tissue Sections Xiaochun Xu and Reuben Lotan In Situ Hybridization with S-Labeled Probes for Retinoid Receptors Karen Niederreither and Pascal Dollé Isolation of Retinoid Receptors from Mammalian Cells Ann K. Daly and Christopher P. F. Redfern Analysis of Retinoid Receptor Phosphorylation Ali Tahayato, Christophe Rachez, Pierre Formstecher, and Philippe Lefebvre Photoaffinity Labeling of RARs and Mapping of Labeled Sites by an Endoproteinase Combination Techniqu Yuichi Hashimoto and Toru Sasaki PCR Cloning of N-Terminal RAR Isoforms and APL-Associated PLZF-RARa Fusion Proteins Arthur Zelent RT-PCR in Diagnosis and Disease Monitoring of Acute Promyelocytic Leukemia (APL) David Grimwade, Stephen Langabeer, Kathy Howe, and Ellen Solomon A Two-Hybrid Protein Interaction System to Identify Factors That Interact with Retinoid and Vitamin D Receptors Paul N. MacDonald Gel-Shift Analysis and Identification of RXREs and RAREs by PCR-Based Selection Myriam I. Baes and Peter E. Declercq Identification and Cloning of RA-Regulated Genes by mRNA Differential Display Jay A. White and Martin Petkovich Gene Targeting of Retinoid Receptors David Lohnes Index

Blurb

Christopher Redfern brings together in Retinoid Protocols a comprehensive collection of key biochemical and molecular methods for retinoid research. These easily reproducible techniques range from methods of handling and analyzing retinoids to advanced protocols for gene targeting, from binding protein function in transgenic animals to the study of nuclear retinoid receptors. In addition, the book makes accessible to all retinoid researchers such advanced techniques as cloning via RT-PCR, fluorimetry methods, recombinant protein purification and characterization, the use of antisense oligonucleotides to study the role of CRABPs, immunological and in situ hybridization methods, photoaffinity labeling, gel-shift analysis, and differential display.

Many of the chapters in Retinoid Protocols represent landmarks in retinoid methodology because they give step-by-step instructions for techniques central to understanding cellular control by retinoids. These indispensable techniques will be fundamental for all those working with retinoids, and so will play a key role in the development of new drugs and therapies for treating human disease.