In vivo techniques to investigate the internalization profi le of opioid receptors

Amynah A. Pradhan, Vivianne L. Tawfik, Alycia F. Tipton, GréGory Scherrer

Research output: Contribution to journalArticlepeer-review

8 Scopus citations


G-protein-coupled receptors (GPCRs) regulate a remarkable diversity of biological functions, and are thus often targeted for drug therapies. Receptor internalization is commonly observed following agonist binding and activation. Receptor trafficking events have been well characterized in cell systems, but the in vivo signifi cance of GPCR internalization is still poorly understood. To address this issue, we have developed an innovative knock-in mouse model, where an opioid receptor is directly visible in vivo. These knockin mice express functional fl uorescent delta opioid receptors (DOR-eGFP) in place of the endogenous receptor, and these receptors are expressed at physiological levels within their native environment. DOR-eGFP mice have proven to be an extraordinary tool in studying receptor neuroanatomy, real-time receptor trafficking in live neurons, and in vivo receptor internalization. We have used this animal model to determine the relationship between receptor trafficking in neurons and receptor function at a behavioral level. Here, we describe in detail the construction and characterization of this knockin mouse. We also outline how to use these mice to examine the behavioral consequences of agonist-specific trafficking at the delta opioid receptor. These techniques are potentially applicable to any GPCR, and highlight the powerful nature of this imaging tool.

Original languageEnglish
Pages (from-to)87-104
Number of pages18
JournalMethods in Molecular Biology
StatePublished - 2015


  • Behavior
  • Delta opioid receptor
  • G-protein-coupled receptor
  • Immunohistochemistry
  • Ligand-directed signaling
  • Mouse
  • Pain
  • Receptor trafficking


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