Heteromerization of μ-opioid receptor and cholecystokinin B receptor through the third transmembrane domain of the μ-opioid receptor contributes to the anti-opioid effects of cholecystokinin octapeptide

다국어 초록 (Multilingual Abstract)

Activation of the cholecystokinin type B receptor (CCKBR) by cholecystokinin octapeptide (CCK-8) inhibits opioid analgesia. Chronic opiate treatment leads to an increase in the CCK-8 concentration and thus enhances the antagonism of CCK-8 against opioid analgesia; the underlying molecular mechanisms remain of great interest. In the present study, we validated the colocalization of the μ-opioid receptor (MOR) and CCKBR in pain signal transmissionrelated spinal cord dorsal horn and dorsal root ganglion neurons of rats. Co-immunoprecipitation (Co-IP) and fluorescence lifetime-imaging-microscopy-based fluorescence resonance energy transfer (FLIM-FRET) assays showed that MOR heteromerized with CCKBR directly in transfected HEK293 cells. Combined with MOR mutant construction, the third transmembrane domain of MOR (TM3MOR) was demonstrated to participate in heteromerization with CCKBR.
Receptor ligand binding, ERK phosphorylation and cAMP assays showed that MOR heteromerization with CCKBR weakened the activity of MOR. A cell-penetrating interfering peptide consisting of TM3MOR and TAT (a transactivator of HIV-1) sequences from the N terminal to the C terminal disrupted the MOR–CCKBR interaction and restored the activity of MOR in transfected HEK293 cells. Furthermore, intrathecal application of the TM3MOR-TAT peptide alleviated CCK-8-injection-induced antagonism to morphine analgesia in rats. These results suggest a new molecular mechanism for CCK-8 antagonism to opioid analgesia in terms of G-protein-coupled receptor (GPCR) interaction through direct heteromerization. Our study may provide a potential strategy for pain management with opioid analgesics.

참고문헌 (Reference)

1 C.T. Dourish, "The selective CCK-B receptor antagonist L-365,260 enhances morphine analgesia and prevents morphine tolerance in the rat" Elsevier BV 176 (176): 35-44, 1990

2 Yu, L., "The role of TRPV1 in different subtypes of dorsal root ganglion neurons in rat chronic inflammatory nociception induced by complete Freund's adjuvant" 4 : 61-, 2008

3 S. Ferre, "Synergistic interaction between adenosine A2A and glutamate mGlu5 receptors: Implications for striatal neuronal function" Proceedings of the National Academy of Sciences 99 (99): 11940-11945, 2002

4 S. Ferre, "Stimulation of high-affinity adenosine A2 receptors decreases the affinity of dopamine D2 receptors in rat striatal membranes." Proceedings of the National Academy of Sciences 88 (88): 7238-7241, 1991

5 Xing Xiao, "Shp-1 dephosphorylates TRPV1 in dorsal root ganglion neurons and alleviates CFA-induced inflammatory pain in rats" Ovid Technologies (Wolters Kluwer Health) 156 (156): 597-608, 2015

6 X Ding, "Reversal of tolerance to morphine but no potentiation of morphine-induced analgesia by antiserum against cholecystokinin octapeptide" Elsevier BV 25 (25): 1155-1160, 1986

7 Travis Omer, "Reduced temporal sampling effect on accuracy of time-domain fluorescence lifetime Förster resonance energy transfer" SPIE-Intl Soc Optical Eng 19 (19): 086023-, 2014

8 Gemma Navarro, "Quaternary structure of a G-protein-coupled receptor heterotetramer in complex with Gi and Gs" Springer Nature 14 (14): 2016

9 Idanpaan-Heikkila, J. J., "Prevention of tolerance to the antinociceptive effects of systemic morphine by a selective cholecystokinin-B receptor antagonist in a rat model of peripheral neuropathy" 282 : 1366-1372, 1997

10 G. W. Pasternak, "Mu Opioids and Their Receptors: Evolution of a Concept" American Society for Pharmacology & Experimental Therapeutics (ASPET) 65 (65): 1257-1317, 2013

11 Herman, B., "Microscopic analysis of fluorescence resonance energy transfer (FRET)" 261 : 351-370, 2004

12 Dang, V. C., "Mechanisms of rapid opioid receptor desensitization, resensitization and tolerance in brain neurons" 165 : 1704-1716, 1704

13 Jia Liu, "Lysine-Specific Demethylase 1 in Breast Cancer Cells Contributes to the Production of Endogenous Formaldehyde in the Metastatic Bone Cancer Pain Model of Rats" Public Library of Science (PLoS) 8 (8): e58957-, 2013

14 Sun, Y., "Localizing protein-protein interactions in living cells using fluorescence lifetime imaging microscopy" 1251 : 83-107, 2015

15 T. F. Shi, "L-364,718 Potentiates Electroacupuncture Analgesia Through Cck-A Receptor of Pain-Related Neurons in the Nucleus Parafascicularis" Springer Nature 36 (36): 129-138, 2011

16 Ji-Sheng Han, "Is cholecystokinin octapeptide (CCK-8) a candidate for endogenous antiopioid substrates?" Elsevier BV 5 (5): 399-402, 1985

17 W XIONG, "Involvement of endogenous cholecystokinin in tolerance to morphine antinociception in the nucleus accumbens of rats" Elsevier BV 173 (173): 116-121, 2006

18 Schoberer, J., "Investigating protein-protein interactions in the plant endomembrane system using multiphoton-induced FRET-FLIM" 1209 : 81-95, 2014

19 Xu Zhang, "Interaction and regulatory functions of μ- and δ-opioid receptors in nociceptive afferent neurons" Springer Nature 28 (28): 121-130, 2012

20 Y. Zhou, "Increased release of immunoreactive CCK-8 by electroacupuncture and enhancement of electroacupuncture analgesia by CCK-B antagonist in rat spinal cord" Elsevier BV 24 (24): 139-144, 1993

21 Yun Hu, "Incorporation of Local Structural Preference Potential Improves Fold Recognition" Public Library of Science (PLoS) 6 (6): e17215-, 2011

22 Bryen A. Jordan, "G-protein-coupled receptor heterodimerization modulates receptor function" Springer Nature 399 (399): 697-700, 1999

23 Matthew B. Dalrymple, "G protein-coupled receptor dimers: Functional consequences, disease states and drug targets" Elsevier BV 118 (118): 359-371, 2008

24 I. Gomes, "G Protein-Coupled Receptor Heteromerization: A Role in Allosteric Modulation of Ligand Binding" American Society for Pharmacology & Experimental Therapeutics (ASPET) 79 (79): 1044-1052, 2011

25 Shao-Qiu He, "Facilitation of μ-Opioid Receptor Activity by Preventing δ-Opioid Receptor-Mediated Codegradation" Elsevier BV 69 (69): 120-131, 2011

26 P. Faris, "Evidence for the neuropeptide cholecystokinin as an antagonist of opiate analgesia" American Association for the Advancement of Science (AAAS) 219 (219): 310-312, 1983

27 Colin T. Dourish, "Enhancement of morphine analgesia and prevention of morphine tolerance in the rat by the cholecystokinin antagonist L-364,718" Elsevier BV 147 (147): 469-472, 1988

28 Fabrizio Benedetti, "Disruption of opioid-induced placebo responses by activation of cholecystokinin type-2 receptors" Springer Nature 213 (213): 791-797, 2011

29 Fang Liu, "Direct protein–protein coupling enables cross-talk between dopamine D5 and γ-aminobutyric acid A receptors" Springer Nature 403 (403): 274-280, 2000

30 Yan Li, "Cholecystokinin-octapeptide antagonizes morphine analgesia in periaqueductal gray of the rat" Elsevier BV 480 (480): 105-110, 1989

31 Chun-Xiao Yang, "Cholecystokinin-8 antagonizes electroacupuncture analgesia through its B receptor in the caudate nucleus" Wiley 13 (13): 93-98, 2010

32 Josimari M. DeSantana, "Cholecystokinin receptors mediate tolerance to the analgesic effect of TENS in arthritic rats" Ovid Technologies (Wolters Kluwer Health) 148 (148): 84-93, 2010

33 Liu, N. J., "Cholecystokinin octapeptide reverses mu-opioid-receptormediated inhibition of calcium current in rat dorsal root ganglion neurons" 275 : 1293-1299, 1995

34 Xiao-Jing Wang, "Cholecystokinin octapeptide antagonized opioid analgesia mediated by μ- and κ- but not δ-receptors in the spinal cord of the rat" Elsevier BV 523 (523): 5-10, 1990

35 J. S. Han, "Cholecystokinin octapeptide (CCK-8): Antagonism to electroacupuncture analgesia and a possible role in electroacupuncture tolerance" Ovid Technologies (Wolters Kluwer Health) 27 (27): 101-115, 1986

36 Su-Fen Pu, "Cholecystokinin octapeptide (CCK-8) antagonizes morphine analgesia in nucleus accumbens of the rat via the CCK-B receptor" Elsevier BV 657 (657): 159-164, 1994

37 Cheng Huang, "CCKB receptor antagonist L365,260 potentiates the efficacy to and reverses chronic tolerance to electroacupuncture-induced analgesia in mice" Elsevier BV 71 (71): 447-451, 2007

38 Li, N., "BN-9, a chimeric peptide with mixed opioid and neuropeptide FF receptor agonistic properties, produces nontolerance-forming antinociception in mice" 173 : 1864-1880, 1864

39 S. Ferre, "An Update on Adenosine A2A-Dopamine D2 Receptor Interactions: Implications for the Function of G Protein-Coupled Receptors" Bentham Science Publishers Ltd. 14 (14): 1468-1474, 2008

40 Ji-Sheng Han, "Acupuncture analgesia: Areas of consensus and controversy" Ovid Technologies (Wolters Kluwer Health) 152 (152): S41-S48, 2011

41 Feng-Yu Liu, "Activation of satellite glial cells in lumbar dorsal root ganglia contributes to neuropathic pain after spinal nerve ligation" Elsevier BV 1427 : 65-77, 2012