Description
Opioid Receptors: Methods and Protocols serves as a comprehensive guide to both key new techniques and established methods for the investigation of genetics, structural biology, transcription, and post-transcriptional events of opioid receptors. Other methods cover the cellular detection and trafficking of opioid receptors in vitro and in vivo. Procedures aimed to investigate signaling pathways modulated by opioid receptors and model systems to study opioid receptor-mediated functions are also included. Finally, methods to assay behavioral effects mediated by opioid receptors are described. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective introductions, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls. Wide-ranging and authoritative, Opioid Receptors: Methods and Protocols will help both experienced and new entrants in this field to carry out their experiments successfully and with new inspiration. Part I: Genetic, Structural Biology, Transcriptional and Post-Transcriptional Analysis of Opioid Receptors 1. Overview of Genetic Analysis of Human Opioid Receptors Santi M. Spampinato 2. Computational Structural Biology of Opioid Receptors Davide Provasi 3. Analysis of Epigenetic Mechanisms Regulating Opioid Receptor Gene Transcription Cheol Kyu Hwang, Yadav Wagley, Ping-Yee Law, Li-Na Wei, and Horace H. Loh 4. Renilla Luciferase Reporter Assay to Study 3’UTR-Driven Post-Transcriptional Regulations of OPRM1 Gabriele Vincelli and Andrea Bedini 5. High-Throughput Gene Expression Profiling of Opioid-Induced Alterations in Discrete Brain Areas Michal Korostynski, Marcin Piechota, Slawomir Golda, and Ryszard Przewlocki Part II: Cellular Detection and Analysis of Opioid Receptors 6. Real-Time Imaging of Mu Opioid Receptors by Total Internal Reflection Fluorescence Microscopy Cristina Roman-Vendrell and Guillermo Ariel Yudowski 7. In Vivo Techniques to Investigate the Internalization Profile of Opioid Receptors Amynah A. Pradhan, Vivianne L. Tawfik, Alycia F. Laboy, and Grgory Scherrer 8. Monitoring Opioid Receptor Interaction in Living Cells by Bioluminescence Resonance Energy Transfer (BRET) Monica Baiul a 9. Bioluminescence Resonance Energy Transfer (BRET) to Detect the Interactions Between Kappa Opioid Receptor and Non-Visual Arrestins Andrea Bedini 10. Identification and Verification of Proteins Interacting with the Kappa Opioid Receptor (KOPR) Chongguang Chen, Peng Huang, and Lee-Yuan Liu-Chen 11. Detection of Mu Opioid Receptor (MOPR) and Its Glycosylation in Rat and Mouse Brains by Western Blot with Anti-C, an Affinity-Purified Polyclonal Anti-MOPR Antibody Peng Huang, Chongguang Chen, and Lee-Yuan Liu-Chen 12. Immunohistochemical Analysis of Opioid Receptors in Peripheral Tissues Yvonne Schmidt and Halina Machelska Part III: Analysis of Signaling Events Modulated by Opioid Receptors 13. [35S]GTPgS-Autoradiography for Studies of Opioid Receptor Functionality Alfhild Grnbladh and Mathias Hallberg 14. Fluorescence-Based, High-Throughput Assays for -Opioid Receptor Activation Using Membrane Potential-Sensitive Dye Alisa Knapman and Mark Connor 15. Analysis of Potassium and Calcium Imaging to Assay the Function of Opioid Receptors Viola Spahn, Dinah Nockemann, and Halina Machelska 16. Electrophysiological Patch Clamp Assay to Monitor the Action of Opioid Receptors Viola Spahn, Dinah Nockemann, a nd Halina Machelska Part IV: Model Systems to Studying Opioid Receptor-Mediated Functions 17. Skin-Nerve Preparation to Assay the Function of Opioid Receptors in Peripheral Endings of Sensory Neurons Rabih Moshourab, Yvonne Schmidt, and Halina Machelska 18. Mechanical Nociception Measurement in Mice and Rats with Automated Von Frey Equipment Gabriele Campana and Roberto Rimondini 19. Detecting Zinc Release Induced by Mu-Opioid Receptor Agonists in Brain Slices Mara Rodrguez-Muoz, Pilar Snchez-Blzquez, Concha Bailn, and Javier Garzn 20. Opioid Receptors: Methods for Detection and Their Modes of Actions in the Eye Shahid Husain 21. Evaluation of Murine Macrophage Cytokine Production After In Vivo Morphine Treatment Silvia Franchi, Mara Castelli, Sarah Moretti, Alberto Panerai, and Paola Sacerdote 22. Measurement of Macrophage Toll Like Receptor 4 Expression After Morphine Treatment Mara Castelli, Alberto Panerai, Paola Sacerdote, and Silvia Franchi 23. The Role of Opioid Receptors in Migration and Wound Recovery In Vitro in Cultured Human Keratinocytes and Fibroblasts Mei Bigliardi-Qi and Paul L. Bigliardi Part V: Behavioral Effects Mediated by Opioid Receptors 24. Role of Opioid Receptors in the Reinstatemen t of Opioid-Seeking Behavior: An Overview Liana Fattore, Paola Fadda, Mary Silvia Antinori, and Walter Fratta 25. Analysis of Opioid-Seeking Reinstatement in the Rat Liana Fattore, Paola Fadda, Mary Tresa Zanda, and Walter Fratta 26. Induction of a High Alcohol Consumption in Rats and Mice: Role of Opioid Receptors Roberto Rimondini and Gabriele Campana 27. Evaluation of Social and Non-Social Behaviors Mediated by Opioids in Mouse Pups Francesca R. D’Amato
