Includes cutting-edge methods and protocols in the study of opioid receptors
Provides step-by-step detail essential for reproducible results
Contains key notes and implementation advice from the experts
Includes cutting-edge methods and protocols in the study of opioid receptors
Provides step-by-step detail essential for reproducible results
Contains key notes and implementation advice from the experts
Includes supplementary material: sn.pub/extras
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, tranillegalscription, and post-tranillegalscriptional 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, Tranillegalscriptional and Post-Tranillegalscriptional 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 Tranillegalscription
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-Tranillegalscriptional 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 Grégory 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-µC, 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 Grönbladh and Mathias Hallberg
14. Fluorescence-Based, High-Throughput Assays for µ-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
María Rodríguez-Muñoz, Pilar Sánchez-Blázquez, Concha Bailón, and Javier Garzón
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
"Book edited by Santi M. Spampinato offers a review of state-of-the-art techniques used to analyze opioid receptors functionality on molecular, cellular, structural, and organism level. ... The book is aimed for practicing researchers actively engaged in the field of opioid receptors investigation. Everyone from graduate students to principal investigator may find it useful.” (Lubica Lacinova, General Physiology and Biophysics, Vol. 34 (3), 2015)
Overview of Genetic Analysis of Human Opioid Receptors.- Computational Structural Biology of Opioid Receptors.- Analysis of Epigenetic Mechanisms Regulating Opioid Receptor Gene Tranillegalscription.- Renilla Luciferase Reporter Assay to Study 3'UTR-Driven Post-Tranillegalscriptional Regulations of OPRM1.- High-Throughput Gene Expression Profiling of Opioid-Induced Alterations in Discrete Brain Areas.- Real-Time Imaging of Mu Opioid Receptors by Total Internal Reflection Fluorescence Microscopy.- In Vivo Techniques to Investigate the Internalization Profile of Opioid Receptors.- Monitoring Opioid Receptor Interaction in Living Cells by Bioluminescence Resonance Energy Transfer (BRET).- Bioluminescence Resonance Energy Transfer (BRET) to Detect the Interactions Between Kappa Opioid Receptor and Non-Visual Arrestins.- Identification and Verification of Proteins Interacting with the Kappa Opioid Receptor (KOPR).- Detection of Mu Opioid Receptor (MOPR) and Its Glycosylation in Rat and Mouse Brains by Western Blot with Anti-µC, an Affinity-Purified Polyclonal Anti-MOPR Antibody.- Immunohistochemical Analysis of Opioid Receptors in Peripheral Tissues.- [35S]GTPgS-Autoradiography for Studies of Opioid Receptor Functionality.- Fluorescence-Based, High-Throughput Assays for µ-Opioid Receptor Activation Using Membrane Potential-Sensitive Dye.- Analysis of Potassium and Calcium Imaging to Assay the Function of Opioid Receptors.- Electrophysiological Patch Clamp Assay to Monitor t
he Action of Opioid Receptors.- Skin-Nerve Preparation to Assay the Function of Opioid Receptors in Peripheral Endings of Sensory Neurons.- Mechanical Nociception Measurement in Mice and Rats with Automated Von Frey Equipment.- Detecting Zinc Release Induced by Mu-Opioid Receptor Agonists in Brain Slices.- Opioid Receptors: Methods for Detection and Their Modes of Actions in the Eye.- Evaluation of Murine Macrophage Cytokine Production After In Vivo Morphine Treatment.- Measurement of Macrophage Toll Like Receptor 4 Expression After Morphine Treatment.- The Role of Opioid Receptors in Migration and Wound Recovery In Vitro in Cultured Human Keratinocytes and Fibroblasts.- Role of Opioid Receptors in the Reinstatement of Opioid-Seeking Behavior: An Overview.- Analysis of Opioid-Seeking Reinstatement in the Rat.- Induction of a High Alcohol Consumption in Rats and Mice: Role of Opioid Receptors.- Evaluation of Social and Non-Social Behaviors Mediated by Opioids in Mouse Pups.
"Book edited by Santi M. Spampinato offers a review of state-of-the-art techniques used to analyze opioid receptors functionality on molecular, cellular, structural, and organism level. ... The book is aimed for practicing researchers actively engaged in the field of opioid receptors investigation. Everyone from graduate students to principal investigator may find it useful." (Lubica Lacinova, General Physiology and Biophysics, Vol. 34 (3), 2015)
Inhaltsverzeichnis
Overview of Genetic Analysis of Human Opioid Receptors.- Computational Structural Biology of Opioid Receptors.- Analysis of Epigenetic Mechanisms Regulating Opioid Receptor Gene Tranillegalscription.- Renilla Luciferase Reporter Assay to Study 3'UTR-Driven Post-Tranillegalscriptional Regulations of OPRM1.- High-Throughput Gene Expression Profiling of Opioid-Induced Alterations in Discrete Brain Areas.- Real-Time Imaging of Mu Opioid Receptors by Total Internal Reflection Fluorescence Microscopy.- In Vivo Techniques to Investigate the Internalization Profile of Opioid Receptors.- Monitoring Opioid Receptor Interaction in Living Cells by Bioluminescence Resonance Energy Transfer (BRET).- Bioluminescence Resonance Energy Transfer (BRET) to Detect the Interactions Between Kappa Opioid Receptor and Non-Visual Arrestins.- Identification and Verification of Proteins Interacting with the Kappa Opioid Receptor (KOPR).- Detection of Mu Opioid Receptor (MOPR) and Its Glycosylation in Rat and Mouse Brains by Western Blot with Anti-µC, an Affinity-Purified Polyclonal Anti-MOPR Antibody.- Immunohistochemical Analysis of Opioid Receptors in Peripheral Tissues.- [35S]GTPgS-Autoradiography for Studies of Opioid Receptor Functionality.- Fluorescence-Based, High-Throughput Assays for µ-Opioid Receptor Activation Using Membrane Potential-Sensitive Dye.- Analysis of Potassium and Calcium Imaging to Assay the Function of Opioid Receptors.- Electrophysiological Patch Clamp Assay to Monitor the Action of Opioid Receptors.- Skin-Nerve Preparation to Assay the Function of Opioid Receptors in Peripheral Endings of Sensory Neurons.- Mechanical Nociception Measurement in Mice and Rats with Automated Von Frey Equipment.- Detecting Zinc Release Induced by Mu-Opioid Receptor Agonists in Brain Slices.- Opioid Receptors: Methods for Detection and Their Modes of Actions in the Eye.- Evaluation of Murine Macrophage Cytokine Production After In Vivo Morphine Treatment.- Measurement of Macrophage Toll Like Receptor 4 Expression After Morphine Treatment.- The Role of Opioid Receptors in Migration and Wound Recovery In Vitro in Cultured Human Keratinocytes and Fibroblasts.- Role of Opioid Receptors in the Reinstatement of Opioid-Seeking Behavior: An Overview.- Analysis of Opioid-Seeking Reinstatement in the Rat.- Induction of a High Alcohol Consumption in Rats and Mice: Role of Opioid Receptors.- Evaluation of Social and Non-Social Behaviors Mediated by Opioids in Mouse Pups.
Klappentext
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, tranillegalscription, and post-tranillegalscriptional 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.nWide-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.
Includes cutting-edge methods and protocols in the study of opioid receptors
rn
Provides step-by-step detail essential for reproducible results
rn
Contains key notes and implementation advice from the experts
rn
