Differential activation and trafficking of micro-opioid receptors in brain slices.
The activation of G protein-coupled receptors results in a cascade of events that include acute signaling, desensitization, and internalization, and it is thought that not all agonists affect each process to the same extent. The early steps in opioid receptor signaling, including desensitization, have been characterized electrophysiologically using brain slice preparations, whereas most previous studies of opioid receptor trafficking have been conducted in heterologous cell models. This study used transgenic mice that express an epitope-tagged (FLAG) micro-opioid receptor (FLAGMOR) targeted to catecholamine neurons by regulatory elements from the tyrosine hydroxylase gene. Brain slices from these mice were used to study tagged MOR receptors in neurons of the locus ceruleus. Activation of the FLAGMOR with [Met5]enkephalin (ME) produced a hyperpolarization that desensitized acutely to the same extent as native MOR in slices from wild-type mice. A series of opioid agonists were then used to study desensitization and receptor trafficking in brain slices, which was monitored with a monoclonal antibody against the FLAG epitope (M1) conjugated to Alexa 594. Three patterns of receptor trafficking and desensitization were observed: 1) ME, etorphine, and methadone resulted in both receptor desensitization and internalization; 2) morphine and oxymorphone caused significant desensitization without evidence for internalization; and 3) oxycodone was ineffective in both processes. These results show that two distinct forms of signaling were differentially engaged depending on the agonist used to activate the receptor, and they support the hypothesis that ligand-specific regulation of opioid receptors occurs in neurons maintained in brain slices from adult animals.
Arttamangkul
Seksiri
S
Vollum Institute, L474, Department of Behavioral Neuroscience, Oregon Health Sciences University, 3181 W Sam Jackson Park Dr., Portland, OR 97239, USA.
Quillinan
Nidia
N
Low
Malcolm J
MJ
von Zastrow
Mark
M
Pintar
John
J
Williams
John T
JT
D000701:Analgesics, Opioid / Q000494:pharmacology; D000818:Animals; D001921:Brain / Q000378:metabolism*; D002614:Chelating Agents / Q000494:pharmacology; D004533:Egtazic Acid / Q000494:pharmacology; D004594:Electrophysiology; D020875:Enkephalin, Ala(2)-MePhe(4)-Gly(5)- / Q000494:pharmacology; D005048:Etorphine / Q000494:pharmacology; D019085:Fluorescent Antibody Technique, Direct; D008125:Locus Coeruleus / Q000166:cytology / Q000378:metabolism; D008691:Methadone / Q000494:pharmacology; D051379:Mice; D018345:Mice, Knockout; D008822:Mice, Transgenic; D009020:Morphine / Q000494:pharmacology; D009474:Neurons / Q000378:metabolism; D010098:Oxycodone / Q000494:pharmacology; D010111:Oxymorphone / Q000494:pharmacology; D021381:Protein Transport / Q000187:drug effects / Q000502:physiology; D011869:Radioligand Assay; D017450:Receptors, Opioid, mu / Q000819:agonists / Q000378:metabolism* / Q000502:physiology*; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D013997:Time Factors
D000701:Analgesics, Opioid; D002614:Chelating Agents; D017450:Receptors, Opioid, mu; D011993:Recombinant Fusion Proteins; D020875:Enkephalin, Ala(2)-MePhe(4)-Gly(5)-; D005048:Etorphine; D004533:Egtazic Acid; D009020:Morphine; D010111:Oxymorphone; D010098:Oxycodone; D008691:Methadone