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All three peroxisome proliferator-activated receptor (PPAR) subtypes share a high degree of structural homology but differ in function, tissue distribution and ligand specificity. PPARs play critical roles as regulators of numerous physiological as well as pathophysiological pathways, and efforts are currently underway to fully characterize their functioning and to develop safer and more effective PPAR modulators to treat a myriad of diseases and conditions. In Peroxisome proliferator-Activated Receptors: Methods and Protocols, renowned experts in the PPAR arena provide detailed protocols for investigating these receptors. Chapters contain methods ranging from the cloning of receptors to their knockdown, to protocols exploring posttranslational modifications of PPARs and coactivators, as well as receptor subcellular localization.
Also assembled are methods to evaluate the involvement of these receptors in behavior functions, an emerging facet in PPAR research. Written in the 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 protocols, and notes on troubleshooting and avoiding known pitfalls. With its well-honed methodologies, Peroxisome proliferator-Activated Receptors: Methods and Protocols, will be a useful resource for all seeking to advance their knowledge of this field.
1. PPARs: History and Advances Jihan Youssef and Mostafa Badr
2. PPAR-Alpha Cloning, Expression and Characterization Suong N. T. Ngo and Ross McKinnon
3. PPARGC1A and PPARG Genotyping. Beginner's Guide to Genotyping with Unlabeled Probes Viera Habalova and Josef Zidzik
4. Generation of an Inducible, Cardiomyocyte-Specific Transgenic Mouse Model with PPARbeta/d Overexpression Teayoum Kim, Olga Zhelyabovska, Jian Liu, and Qinglin Yang
5. Specific knockdown of PPARd Gene in Colon Cancer Cells by Lentivirus-Mediated RNA Interfering Lie Yang, Zong-Guang Zhou, and Xiao-Feng Sun
6. Dominant-Negative and Knockdown Approaches to Studying PPAR activity Sudhir Singh and Robert G. Bennett
7. Producing PPARgamma2 Knockdown in Mouse Liver Tomomi Yamazaki and Osamu Ezaki
8. Adipose Tissue-Specific PPARg Gene Targeting Weimin He
9. Site-Directed Mutagenesis to Study the Role of Specific Amino Acids in the Ligand Binding Domain of PPARs Nico Mitro, Frederica Gilardi, Marco Giudici, Cristina Godio, Elena Scotti, and Maurizio Crestani
10. PPAR SUMOylation: Some Useful Experimental Tips Benoit Pourcet, Bart Staels, and Corine Glineur
11. Analyzing Phosphorylation-Dependent Regulation of Subcellular Transcriptional Activity of Transcriptional Coactivator NT-PGC-1a Ji Suk Chang and Thomas W. Gettys
12. In vivo Studies of PPAR-Chromatin Interactions: Chromatin Immunoprecipitation for Single-Locus and Genomewide Analyses Till Adhikary and Rolf Muller
13. FISH Analysis Using PPARg Specific Probes for Detection of PAX8-PPARg Translocation in Follicular Thyroid Neoplasms Sharifah Noor Akmal Syed Hussain, Chia Wai Kit, and Zubaidah Zakaraia
14. Immunohistochemical Techniques to Identify and Localize Proteins of Interest in Paraffin-Embedded Tissue Sections Carolyn M. Komar and Mary Jane Long
15. Determination of PPAR Expression by Western-Blot Aziz Gullich and Catherine Coirault
16. Fluorescence Resonance Energy Transfer Techniques to Study Ligand-Mediated Interactions of PPARs with Coregulators Nico Mitro, Cristina Godio, and Maurizio Crestani
17. Estimation of the PPARa Agonism of Fibrates by a Combined MM-Docking Approach Fabio Lannutti, Alessandro Marrone, and Nazzareno Re
18. Combined Biophysical and Cell-based Approaches for the Assessment of Ligand Binding to PPARg Silvia Zorrilla and Dolores Perez-Sala
19. Exploring PPAR Modulation in Experimental Mice Anna Rull, Gerard Aragones, Raul Beltran-Debon, Esther Rodriguez-Gallego, Jordi Camps, Jorge Joven
20. Induction of Adipogenic Differentiation in Three-Dimensional Culture Model on a Novel Microfabricated Scaffold Yoshitaka Miyagawa, Hajime Okita, and Nobutaka Kiyokawa
21. Analyzing PPAa/Ligand Interactions by Chemical Cross-Linking and High-Resolution Mass Spectrometry Mathias Muller, Christian Ihling, and Andrea Sinz
22. Synthesis, Mass Spectrometric Characterization, and Analysis of the PPARd Agonist GW1516 and its Major Human Metabolites - Targets in Sports Drug Testing Mario Thevis, Ines Moller, Simon Beuk, and Wihelm Schanzer
23. LC-MS-Based Method for the Qualitative and Quantitative Analysis of the Novel PPARg Agonist KR-62980 Mi-Sun Kim and Myung Ae Bae
24. Behavioral Paradigms to Evaluate PPAR Modulation in Animal Models of Brain Injury Dana Greene-Schloesser, Caroline I. Schnegg, and Mike E. Robbins
