Temporal dependence of shifts in mu opioid receptor mobility at the cell surface after agonist binding observed by single-particle tracking
| Autor: | Diego Krapf, Marissa J. Metz, Reagan L. Pennock, Shane T. Hentges |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 2018 |
| Předmět: |
0301 basic medicine
Time Factors Intravital Microscopy Enkephalin Receptors Opioid mu lcsh:Medicine Mice chemistry.chemical_compound 0302 clinical medicine polycyclic compounds Internalization lcsh:Science Receptor media_common 0303 health sciences education.field_of_study Microscopy Confocal Multidisciplinary biology Single Molecule Imaging DAMGO Signal transduction μ-opioid receptor Cell signalling Signal Transduction Agonist medicine.drug_class media_common.quotation_subject Population Clathrin Article 03 medical and health sciences Single-molecule biophysics Cell Line Tumor Quantum Dots mental disorders medicine Animals education 030304 developmental biology Cell Membrane lcsh:R Colocalization Enkephalin Ala(2)-MePhe(4)-Gly(5) 030104 developmental biology Microscopy Fluorescence chemistry nervous system biology.protein Biophysics Feasibility Studies lcsh:Q human activities 030217 neurology & neurosurgery |
| Zdroj: | Scientific Reports, Vol 9, Iss 1, Pp 1-13 (2019) Scientific Reports |
| DOI: | 10.1101/402826 |
| Popis: | Agonist binding to the mu opioid receptor (MOR) results in conformational changes that allow recruitment of G-proteins, activation of downstream effectors and eventual desensitization and internalization, all of which could affect receptor mobility. The present study employed single particle tracking (SPT) of quantum dot labeled FLAG-tagged MORs to examine shifts in MOR mobility after agonist binding. FLAG-MORs on the plasma membrane were in both mobile and immobile states under basal conditions. Activation of FLAG-MORs with DAMGO caused an acute increase in the fraction of mobile MORs, and free portions of mobile tracks were partially dependent on interactions with G-proteins. In contrast, 10-minute exposure to DAMGO or morphine increased the fraction of immobile FLAG-MORs. While the decrease in mobility with prolonged DAMGO exposure corresponded to an increase in colocalization with clathrin, the increase in colocalization was present in both mobile and immobile FLAG-MORs. Thus, no single mobility state of the receptor accounted for colocalization with clathrin. These findings demonstrate that SPT can be used to track agonist-dependent changes in MOR mobility over time, but that the mobility states observed likely arise from a diverse set of interactions and will be most informative when examined in concert with particular downstream effectors. |
| Databáze: | OpenAIRE |
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