Main description:

This second edition presents an up-to-date chapters describing the most relevant and novel techniques employed to study the opioid receptors. Chapters detail transcriptional and post-transcriptional analysis, cellular detection of opioid receptors, analysis of signaling events modulated by opioid receptors, model systems to studying opioid receptor-mediated functions, and behavioral effects mediated by opioid receptors. Written in the highly 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 laboratory protocols, and tips on troubleshooting and avoiding known pitfalls.

Authoritative and cutting-edge, Opioid Receptors: Methods and Protocols, Second Edition aims to ensure successful results in the further study of this vital field.

Contents:

Part I: transcriptional and post-transcriptional analysis and cellular detection of opioid receptors

1. Overview of Genetic Analysis of Human Opioid Receptors

Santi M. Spampinato

2. Renilla luciferase Reporter Assay to Study 3'UTR-driven post-Transcriptional Regulation of OPRM1

Gabriele Vincelli and Andrea Bedini

3. Fluorescence Colocalization Analysis of Cellular Distribution of MOR-1

Vittoria Borgonetti and Nicoletta Galeotti

4. Monitoring Opioid Receptors Interaction in Living Cells by Bioluminescence Resonance Energy Transfer (BRET)

Monica Baiula

5. Bioluminescence Resonance Energy Transfer (BRET) to Detect the Interactions Between Kappa Opioid Receptor and Non-visual Arrestins

Andrea Bedini

6. Functionalization and Bioconjugation of Nanoruby for Long-term, Ultrasensitive Imaging of -Opioid Receptors

Rashmi Pillai , Mark Connor , and Varun Sreenivasan

7. Immunohistochemical Analysis of Opioid Receptors in Peripheral Tissues

Yvonne Schmidt and Halina Machelska

8. Real-Time Quantitative Reverse Transcription PCR for Detection of Opioid Receptors in Immune Cells Melih OEzgur Celik, Dominika Labuz, and Halina Machelska

Part II: Analysis of signaling events modulated by opioid receptors

9. Quantitative Analysis of MOR-1 Internalization in Spinal Cord of Morphine-tolerant Mice

Vittoria Borgonetti and Nicoletta Galeotti

10. GTPgS-autoradiography for Studies of Opioid Receptor Functionality

Alfhild Groenbladh and Mathias Hallberg

11. Evaluating Opioid Mediated Adenylyl Cyclase Inhibition in Live Cells using a BRET-based Assay

Preeti Manandhar, Shivani Sachdev and Marina Santiago

12. Patch Clamp Analysis of Opioid-Induced Kir3 Currents in Mouse Peripheral Sensory Neurons Following Nerve Injury

Viola Seitz, Philip Stoetzner, Dominika Labuz, and Halina Machelska

13. Opioid Modulation of Neuronal Iron and Potential Contributions to NeuroHIV

Bradley Nash, Elena Irollo, Renato Brandimarti, and Olimpia Meucci

14. An In Vitro Assay to Study the Role of Opioids in Modulating Immune Cell Adhesion

Monica Baiula

15. Assessing Cell Viability Effects of Opioids in Primary Cortical Cells from Rat

Alfhild Groenbladh, Erik Nylander, Sofia Zelleroth, and Mathias Hallberg

16. The Quest for More Effective Analgesics with Reduced Abuse Liability and Fewer Adverse Effects: Promises, Pitfalls and Future Perspectives of Biased Agonists at Opioid Receptors

Andrea Bedini

Part III: Model systems to studying opioid receptor-mediated functions

17. Mechanical Nociception in Mice and Rats: Measurement with Automated von Frey Equipment

Gabriele Campana and Roberto Rimondini

18. Evaluation of Murine Macrophage Cytokine Production after In Vivo Morphine Treatment

Silvia Franchi, Mara Castelli, Sarah Moretti, Alberto Panerai, and Paola Sacerdote

19. Measurement of Macrophage Toll like Receptor 4 Expression After Morphine
Treatment

Mara Castelli, Alberto Panerai, Paola Sacerdote, and Silvia Franchi

Part IV Behavioral effects mediated by opioid receptors

20. Conditioned Place Preference (CPP) in Rats: From Conditioning to Reinstatement Test

Maria Scherma, Liana Fattore,Walter Fratta, and Paola Fadda

21. Analysis of Opioid-Seeking Behavior through the Intravenous Self-administration Reinstatement Model in Rats

Liana Fattore, Paola Fadda, Mary Tresa Zanda, and Walter Fratta

22. Induction of a High Alcohol Consumption in Rats and Mice: Role of Opioid Receptors in Rats and Mice

Roberto Rimondini and Gabriele Campana

23. Early Life Stress as a Probe to Study the Opioid System in Developing Rodents

Stefano Loizzo and Alberto Loizzo

24. Evaluation of m--opioid System Functionality in Mouse Pups: Ultrasonic Vocalizations as an Index Of Infant Attachment

Francesca R. D'Amato