title: Opioid receptor internalization contributes to dermorphin-mediated antinociception.
abstract: Microinjection of opioids into the ventrolateral periaqueductal gray (vlPAG) produces antinociception in part by binding to mu-opioid receptors (MOPrs). Although both high and low efficacy agonists produce antinociception, low efficacy agonists such as morphine produce limited MOPr internalization suggesting that MOPr internalization and signaling leading to antinociception are independent. This hypothesis was tested in awake, behaving rats using DERM-A594, a fluorescently labeled dermorphin analog, and internalization blockers. Microinjection of DERM-A594 into the vlPAG produced both antinociception and internalization of DERM-A594. Administration of the irreversible opioid receptor antagonist beta-chlornaltrexamine (beta-CNA) prior to DERM-A594 microinjection reduced both the antinociceptive effect and the number of DERM-A594 labeled cells demonstrating that both effects are opioid receptor-mediated. Pretreatment with the internalization blockers dynamin dominant-negative inhibitory peptide (dynamin-DN) and concanavalinA (ConA) attenuated both DERM-A594 internalization and antinociception. Microinjection of dynamin-DN and ConA also decreased the antinociceptive potency of the unlabeled opioid agonist dermorphin when microinjected into the vlPAG as demonstrated by rightward shifts in the dose-response curves. In contrast, administration of dynamin-DN had no effect on the antinociceptive effect of microinjecting the GABA(A) receptor antagonist bicuculline into the vlPAG. The finding that dermorphin-induced antinociception is attenuated by blocking receptor internalization indicates that key parts of opioid receptor-mediated signaling depend on internalization.
authors.affiliation: Department of Psychology, Washington State University Vancouver, 14204 NE Salmon Creek Avenue, Vancouver, WA 98686, USA.
authors.forename: T A
authors.initials: TA
authors.lastname: Macey
authors.forename: S L
authors.initials: SL
authors.lastname: Ingram
authors.forename: E N
authors.initials: EN
authors.lastname: Bobeck
authors.forename: D M
authors.initials: DM
authors.lastname: Hegarty
authors.forename: S A
authors.initials: SA
authors.lastname: Aicher
authors.forename: S
authors.initials: S
authors.lastname: Arttamangkul
authors.forename: M M
authors.initials: MM
authors.lastname: Morgan
mesh_terms: D000701:Analgesics, Opioid / Q000737:chemistry / Q000494:pharmacology* / Q000627:therapeutic use; D000818:Animals; D001640:Bicuculline / Q000494:pharmacology; D003208:Concanavalin A / Q000494:pharmacology; D034281:Dynamins / Q000037:antagonists & inhibitors; D005456:Fluorescent Dyes / Q000737:chemistry; D058787:GABA-A Receptor Antagonists; D008297:Male; D008845:Microinjections; D009271:Naltrexone / Q000031:analogs & derivatives / Q000494:pharmacology; D009474:Neurons / Q000378:metabolism; D018847:Opioid Peptides / Q000737:chemistry / Q000494:pharmacology* / Q000627:therapeutic use; D010146:Pain / Q000188:drug therapy* / Q000378:metabolism / Q000503:physiopathology; D010147:Pain Measurement; D010455:Peptides / Q000494:pharmacology; D010487:Periaqueductal Gray / Q000187:drug effects / Q000378:metabolism; D051381:Rats; D017207:Rats, Sprague-Dawley; D017450:Receptors, Opioid, mu / Q000037:antagonists & inhibitors / Q000378:metabolism*
chemical_list: D000701:Analgesics, Opioid; D005456:Fluorescent Dyes; D058787:GABA-A Receptor Antagonists; D018847:Opioid Peptides; D010455:Peptides; D017450:Receptors, Opioid, mu; D003208:Concanavalin A; C030058:dermorphin; D009271:Naltrexone; C017120:chlornaltrexamine; D034281:Dynamins; D001640:Bicuculline
references.citation: J Biol Chem. 2002 Sep 13;277(37):33783-90
references.pmid: 12097326
references.citation: J Neurosci. 1999 Jan 1;19(1):56-63
references.pmid: 9870938
references.citation: J Neurosci. 2002 Jul 1;22(13):5769-76
references.pmid: 12097530
references.citation: J Neurosci. 2006 Apr 12;26(15):4118-25
references.pmid: 16611829
references.citation: Mol Pharmacol. 1995 Apr;47(4):666-76
references.pmid: 7723728
references.citation: Brain Res. 1991 Sep 6;558(2):224-30
references.pmid: 1664270
references.citation: Brain Res. 1976 Feb 27;103(3):501-13
references.pmid: 1252940
references.citation: Mol Pharmacol. 2005 Oct;68(4):1127-32
references.pmid: 16020743
references.citation: Mol Pharmacol. 2005 Jan;67(1):280-7
references.pmid: 15475572
references.citation: J Neurosci Methods. 2008 May 30;170(2):285-93
references.pmid: 18377995
references.citation: J Pain. 2000 Sep;1(3 Suppl):57-65
references.pmid: 14622844
references.citation: J Biol Chem. 1996 Aug 9;271(32):19021-4
references.pmid: 8702570
references.citation: Mol Cell Neurosci. 2007 Jul;35(3):456-69
references.pmid: 17513124
references.citation: Brain Res. 1989 Oct 30;501(1):116-28
references.pmid: 2572306
references.citation: Proc Natl Acad Sci U S A. 2009 Oct 20;106(42):17615-22
references.pmid: 19822761
references.citation: Br J Pharmacol. 2008 Mar;153 Suppl 1:S379-88
references.pmid: 18059321
references.citation: J Neurophysiol. 1992 Oct;68(4):1027-45
references.pmid: 1432065
references.citation: J Biol Chem. 2003 May 23;278(21):18776-84
references.pmid: 12642578
references.citation: Neuropharmacology. 2000 Jul 24;39(10):1707-19
references.pmid: 10884553
references.citation: Biochem J. 1989 Oct 15;263(2):533-8
references.pmid: 2556997
references.citation: Eur J Biochem. 1990 May 20;189(3):625-35
references.pmid: 2161761
references.citation: Curr Biol. 2008 Jan 22;18(2):129-35
references.pmid: 18207746
references.citation: Mol Pharmacol. 2008 Oct;74(4):972-9
references.pmid: 18612077
references.citation: Pain. 2009 Dec 15;147(1-3):210-6
references.pmid: 19796879
references.citation: Science. 1999 Dec 24;286(5449):2495-8
references.pmid: 10617462
references.citation: Mol Pharmacol. 2008 Apr;73(4):1301-8
references.pmid: 18198283
references.citation: Behav Brain Res. 2005 Oct 14;164(1):61-6
references.pmid: 16029902
references.citation: Traffic. 2007 May;8(5):462-70
references.pmid: 17376169
references.citation: Neuroscience. 2005;135(1):227-34
references.pmid: 16084660
references.citation: Neuron. 2001 Dec 6;32(5):829-39
references.pmid: 11738029
references.citation: J Neurosci. 2005 Mar 23;25(12):3192-8
references.pmid: 15788776
references.citation: Pain. 1995 Sep;62(3):287-293
references.pmid: 8657428
references.citation: J Pharmacol Exp Ther. 2009 Nov;331(2):412-8
references.pmid: 19684256
references.citation: Brain Res. 1994 Nov 28;665(1):85-93
references.pmid: 7882023
references.citation: Cell. 2002 Jan 25;108(2):271-82
references.pmid: 11832216
references.citation: Traffic. 2007 Jul;8(7):848-66
references.pmid: 17587406
references.citation: Curr Opin Neurobiol. 2003 Jun;13(3):348-53
references.pmid: 12850220
references.citation: Mol Pharmacol. 2000 Dec;58(6):1570-80
references.pmid: 11093798
references.citation: J Biol Chem. 2006 Nov 10;281(45):34515-24
references.pmid: 16982618
references.citation: J Pharmacol Exp Ther. 2001 Sep;298(3):865-72
references.pmid: 11504778
references.citation: Brain Res. 2005 Jun 14;1047(1):65-71
references.pmid: 15878767
references.citation: PLoS One. 2009;4(5):e5425
references.pmid: 19412545
references.citation: Neuropharmacology. 2007 Oct;53(5):664-76
references.pmid: 17845806
references.citation: Brain Res. 2008 Mar 4;1197:85-93
references.pmid: 18207137
references.citation: J Neurosci. 2004 Sep 1;24(35):7699-706
references.pmid: 15342737
references.citation: Pharmacol Ther. 2008 Feb;117(2):199-206
references.pmid: 18076994
references.citation: Mol Pharmacol. 2008 Jan;73(1):178-90
references.pmid: 17947509
references.citation: J Biol Chem. 2003 Nov 14;278(46):45978-86
references.pmid: 12939277
references.citation: J Neurosci. 2003 Nov 19;23(33):10515-20
references.pmid: 14627635
references.citation: Naunyn Schmiedebergs Arch Pharmacol. 1982 Mar;318(4):368-9
references.pmid: 7078669
references.citation: Nat Rev Mol Cell Biol. 2009 Sep;10(9):609-22
references.pmid: 19696798
references.citation: Brain Res. 1988 Apr 26;447(1):25-34
references.pmid: 2838129
references.citation: Neural Plast. 2009;2009:516328
references.pmid: 19266049
references.citation: J Biol Chem. 1997 Dec 12;272(50):31648-56
references.pmid: 9395506
references.citation: Mol Endocrinol. 2004 Mar;18(3):640-52
references.pmid: 14684845
references.citation: Neuroscience. 2004;125(3):541-3
references.pmid: 15099667
references.citation: Br J Pharmacol. 2009 Sep;158(1):154-6
references.pmid: 19719778
references.citation: Pharmacol Biochem Behav. 2006 Sep;85(1):214-9
references.pmid: 16979226
references.citation: J Comp Neurol. 2003 Sep 15;464(2):197-207
references.pmid: 12898612
references.citation: Brain Res. 1998 Aug 31;804(1):159-66
references.pmid: 9729359
references.citation: Mol Pharmacol. 2004 Jul;66(1):106-12
references.pmid: 15213301