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Modulation of Gpr39, a G-protein coupled receptor associated with alcohol use in non-human primates, curbs ethanol intake in mice

Neuropsychopharmacology (2019) | Download Citation

Abstract

Alcohol use disorder (AUD) is a chronic condition with devastating health and socioeconomic effects. Still, pharmacotherapies to treat AUD are scarce. In a prior study aimed at identifying novel AUD therapeutic targets, we investigated the DNA methylome of the nucleus accumbens core (NAcc) of rhesus macaques after chronic alcohol use. The G-protein coupled receptor 39 (GPR39) gene was hypermethylated and its expression downregulated in heavy alcohol drinking macaques. GPR39 encodes a Zn2+-binding metabotropic receptor known to modulate excitatory and inhibitory neurotransmission, the balance of which is altered in AUD. These prior findings suggest that a GPR39 agonist would reduce alcohol intake. Using a drinking-in-the-dark two bottle choice (DID-2BC) model, we showed that an acute 7.5 mg/kg dose of the GPR39 agonist, TC-G 1008, reduced ethanol intake in mice without affecting total fluid intake, locomotor activity or saccharin preference. Furthermore, repeated doses of the agonist prevented ethanol escalation in an intermittent access 2BC paradigm (IA-2BC). This effect was reversible, as ethanol escalation followed agonist “wash out”. As observed during the DID-2BC study, a subsequent acute agonist challenge during the IA-2BC procedure reduced ethanol intake by ~47%. Finally, Gpr39 activation was associated with changes in Gpr39 and Bdnf expression, and in glutamate release in the NAcc. Together, our findings suggest that GPR39 is a promising target for the development of prevention and treatment therapies for AUD.

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References

  1. 1.

    SAMHSA. Substance Abuse and Mental Health Services Administration (SAMHSA). National Survey on Drug Use and Health (NSDUH). Table 5.8B—Substance dependence or abuse in the past year among persons aged 18 or older, by demographic characteristics: Percentages, 2013 and 2014.

  2. 2.

    Sinha R. The role of stress in addiction relapse. Curr Psychiatry Rep. 2007;9:388–95.

  3. 3.

    Cervera-Juanes R, Wilhelm LJ, Park B, Grant KA, Ferguson B. Alcohol-dose-dependent DNA methylation and expression in the nucleus accumbens identifies coordinated regulation of synaptic genes. Transl Psychiatry. 2017;7:e994.

  4. 4.

    Holst B, Holliday ND, Bach A, Elling CE, Cox HM, Schwartz TW. Common structural basis for constitutive activity of the ghrelin receptor family. J Biol Chem. 2004;279:53806–17.

  5. 5.

    Holst B, Egerod KL, Schild E, Vickers SP, Cheetham S, Gerlach LO, et al. GPR39 signaling is stimulated by zinc ions but not by obestatin. Endocrinology. 2007;148:13–20.

  6. 6.

    Yasuda S, Miyazaki T, Munechika K, Yamashita M, Ikeda Y, Kamizono A. Isolation of Zn2+ as an endogenous agonist of GPR39 from fetal bovine serum. J Recept Signal Transduct Res. 2007;27:235–46.

  7. 7.

    Depoortere I. GI functions of GPR39: novel biology. Curr Opin Pharmacol. 2012;12:647–52.

  8. 8.

    Dittmer S, Sahin M, Pantlen A, Saxena A, Toutzaris D, Pina AL, et al. The constitutively active orphan G-protein-coupled receptor GPR39 protects from cell death by increasing secretion of pigment epithelium-derived growth factor. J Biol Chem. 2008;283:7074–81.

  9. 9.

    Holst B, Egerod KL, Jin C, Petersen PS, Ostergaard MV, Hald J, et al. G protein-coupled receptor 39 deficiency is associated with pancreatic islet dysfunction. Endocrinology. 2009;150:2577–85.

  10. 10.

    Moechars D, Depoortere I, Moreaux B, de Smet B, Goris I, Hoskens L, et al. Altered gastrointestinal and metabolic function in the GPR39-obestatin receptor-knockout mouse. Gastroenterology. 2006;131:1131–41.

  11. 11.

    Cohen L, Sekler I, Hershfinkel M. The zinc sensing receptor, ZnR/GPR39, controls proliferation and differentiation of colonocytes and thereby tight junction formation in the colon. Cell Death Dis. 2014;5:e1307.

  12. 12.

    Mlyniec K, Budziszewska B, Reczynski W, Sowa-Kucma M, Nowak G. The role of the GPR39 receptor in zinc deficient-animal model of depression. Behav Brain Res. 2013;238:30–35.

  13. 13.

    Mlyniec K, Doboszewska U, Szewczyk B, Sowa-Kucma M, Misztak P, Piekoszewski W, et al. The involvement of the GPR39-Zn(2+)-sensing receptor in the pathophysiology of depression. Studies in rodent models and suicide victims. Neuropharmacology. 2014;79:290–7.

  14. 14.

    Chorin E, Vinograd O, Fleidervish I, Gilad D, Herrmann S, Sekler I, et al. Upregulation of KCC2 activity by zinc-mediated neurotransmission via the mZnR/GPR39 receptor. J Neurosci. 2011;31:12916–26.

  15. 15.

    Perez-Rosello T, Anderson CT, Ling C, Lippard SJ, Tzounopoulos T. Tonic zinc inhibits spontaneous firing in dorsal cochlear nucleus principal neurons by enhancing glycinergic neurotransmission. Neurobiol Dis. 2015;81:14–19.

  16. 16.

    Han Y, Wu SM. Modulation of glycine receptors in retinal ganglion cells by zinc. Proc Natl Acad Sci USA. 1999;96:3234–8.

  17. 17.

    Herin GA, Aizenman E. Amino terminal domain regulation of NMDA receptor function. Eur J Pharmacol. 2004;500:101–11.

  18. 18.

    Hosie AM, Dunne EL, Harvey RJ, Smart TG. Zinc-mediated inhibition of GABA(A) receptors: discrete binding sites underlie subtype specificity. Nat Neurosci. 2003;6:362–9.

  19. 19.

    Lynch JW, Jacques P, Pierce KD, Schofield PR. Zinc potentiation of the glycine receptor chloride channel is mediated by allosteric pathways. J Neurochem. 1998;71:2159–68.

  20. 20.

    Paoletti P, Ascher P, Neyton J. High-affinity zinc inhibition of NMDA NR1-NR2A receptors. J Neurosci. 1997;17:5711–25.

  21. 21.

    Swardfager W, Herrmann N, McIntyre RS, Mazereeuw G, Goldberger K, Cha DS, et al. Potential roles of zinc in the pathophysiology and treatment of major depressive disorder. Neurosci Biobehav Rev. 2013;37:911–29.

  22. 22.

    Mlyniec K, Nowak G. GPR39 up-regulation after selective antidepressants. Neurochem Int. 2013;62:936–9.

  23. 23.

    Mlyniec K, Budziszewska B, Holst B, Ostachowicz B, Nowak G. GPR39 (zinc receptor) knockout mice exhibit depression-like behavior and CREB/BDNF down-regulation in the hippocampus. Int J Neuropsychopharmacol. 2014;23:1–8.

  24. 24.

    Mlyniec K, Starowicz G, Gawel M, Frackiewicz E, Nowak G. Potential antidepressant-like properties of the TC G-1008, a GPR39 (zinc receptor) agonist. J Affect Disord. 2016;201:179–84.

  25. 25.

    Pandey SC. A critical role of brain-derived neurotrophic factor in alcohol consumption. Biol Psychiatry. 2016;79:427–9.

  26. 26.

    Peukert S, Hughes R, Nunez J, He G, Yan Z, Jain R, et al. Discovery of 2-Pyridylpyrimidines as the first orally bioavailable GPR39 agonists. ACS Med Chem Lett. 2014;5:1114–8.

  27. 27.

    Sato N, Yamabuki T, Takano A, Koinuma J, Aragaki M, Masuda K, et al. Wnt inhibitor Dickkopf-1 as a target for passive cancer immunotherapy. Cancer Res. 2010;70:5326–36.

  28. 28.

    Ford MM, Nickel JD, Kaufman MN, Finn DA. Null mutation of 5alpha-reductase type I gene alters ethanol consumption patterns in a sex-dependent manner. Behav Genet. 2015;45:341–53.

  29. 29.

    Thiele TE, Navarro M. “Drinking in the dark” (DID) procedures: a model of binge-like ethanol drinking in non-dependent mice. Alcohol. 2014;48:235–41.

  30. 30.

    Finn DA, Beckley EH, Kaufman KR, Ford MM. Manipulation of GABAergic steroids: sex differences in the effects on alcohol drinking- and withdrawal-related behaviors. Horm Behav. 2010;57:12–22.

  31. 31.

    Hwa LS, Chu A, Levinson SA, Kayyali TM, DeBold JF, Miczek KA. Persistent escalation of alcohol drinking in C57BL/6J mice with intermittent access to 20% ethanol. Alcohol Clin Exp Res. 2011;35:1938–47.

  32. 32.

    Simms JA, Steensland P, Medina B, Abernathy KE, Chandler LJ, Wise R, et al. Intermittent access to 20% ethanol induces high ethanol consumption in Long-Evans and Wistar rats. Alcohol Clin Exp Res. 2008;32:1816–23.

  33. 33.

    Ford MM, Steele AM, McCracken AD, Finn DA, Grant KA. The relationship between adjunctive drinking, blood ethanol concentration and plasma corticosterone across fixed-time intervals of food delivery in two inbred mouse strains. Psychoneuroendocrinology. 2013;38:2598–610.

  34. 34.

    Colombo G, Serra S, Brunetti G, Atzori G, Pani M, Vacca G, et al. The GABA(B) receptor agonists baclofen and CGP 44532 prevent acquisition of alcohol drinking behaviour in alcohol-preferring rats. Alcohol Alcohol. 2002;37:499–503.

  35. 35.

    Ford MM, Yoneyama N, Strong MN, Fretwell A, Tanchuck M, Finn DA. Inhibition of 5alpha-reduced steroid biosynthesis impedes acquisition of ethanol drinking in male C57BL/6J mice. Alcohol Clin Exp Res. 2008;32:1408–16.

  36. 36.

    Serra S, Carai MA, Brunetti G, Gomez R, Melis S, Vacca G, et al. The cannabinoid receptor antagonist SR 141716 prevents acquisition of drinking behavior in alcohol-preferring rats. Eur J Pharmacol. 2001;430:369–71.

  37. 37.

    Stolerman I. Drugs of abuse: behavioural principles, methods and terms. Trends Pharmacol Sci. 1992;13:170–6.

  38. 38.

    Dawson DA, Pulay AJ, Grant BF. A comparison of two single-item screeners for hazardous drinking and alcohol use disorder. Alcohol Clin Exp Res. 2010;34:364–74.

  39. 39.

    Allen DC, Gonzales SW, Grant KA. Effect of repeated abstinence on chronic ethanol self-administration in the rhesus monkey. Psychopharmacology. 2018;235:109–20.

  40. 40.

    Hershfinkel M. The Zinc Sensing Receptor, ZnR/GPR39, in Health and Disease. Int J Mol Sci. 2018;40:439–57.

  41. 41.

    Mlyniec K, Singewald N, Holst B, Nowak G. GPR39 Zn(2+)-sensing receptor: a new target in antidepressant development? J Affect Disord. 2015;174:89–100.

  42. 42.

    Sasi M, Vignoli B, Canossa M, Blum R. Neurobiology of local and intercellular BDNF signaling. Pflug Arch. 2017;469:593–610.

  43. 43.

    Logrip ML, Janak PH, Ron D. Escalating ethanol intake is associated with altered corticostriatal BDNF expression. J Neurochem. 2009;109:1459–68.

  44. 44.

    Perez-Rosello T, Anderson CT, Schopfer FJ, Zhao Y, Gilad D, Salvatore SR, et al. Synaptic Zn2+ inhibits neurotransmitter release by promoting endocannabinoid synthesis. J Neurosci. 2013;33:9259–72.

  45. 45.

    Levine ES, Crozier RA, Black IB, Plummer MR. Brain-derived neurotrophic factor modulates hippocampal synaptic transmission by increasing N-methyl-D-aspartic acid receptor activity. Proc Natl Acad Sci USA. 1998;95:10235–9.

  46. 46.

    Levine ES, Kolb JE. Brain-derived neurotrophic factor increases activity of NR2B-containing N-methyl-D-aspartate receptors in excised patches from hippocampal neurons. J Neurosci Res. 2000;62:357–62.

  47. 47.

    Lin SY, Wu K, Levine ES, Mount HT, Suen PC, Black IB. BDNF acutely increases tyrosine phosphorylation of the NMDA receptor subunit 2B in cortical and hippocampal postsynaptic densities. Brain Res Mol Brain Res. 1998;55:20–27.

  48. 48.

    Suen PC, Wu K, Levine ES, Mount HT, Xu JL, Lin SY, et al. Brain-derived neurotrophic factor rapidly enhances phosphorylation of the postsynaptic N-methyl-D-aspartate receptor subunit 1. Proc Natl Acad Sci USA. 1997;94:8191–5.

  49. 49.

    Minichiello L. TrkB signalling pathways in LTP and learning. Nat Rev Neurosci. 2009;10:850–60.

  50. 50.

    Lessmann V, Heumann R. Modulation of unitary glutamatergic synapses by neurotrophin-4/5 or brain-derived neurotrophic factor in hippocampal microcultures: presynaptic enhancement depends on pre-established paired-pulse facilitation. Neuroscience. 1998;86:399–413.

  51. 51.

    Takei N, Numakawa T, Kozaki S, Sakai N, Endo Y, Takahashi M, et al. Brain-derived neurotrophic factor induces rapid and transient release of glutamate through the non-exocytotic pathway from cortical neurons. J Biol Chem. 1998;273:27620–4.

  52. 52.

    Moykkynen T, Korpi ER. Acute effects of ethanol on glutamate receptors. Basic Clin Pharmacol Toxicol. 2012;111:4–13.

  53. 53.

    Lovinger DM, Roberto M. Synaptic effects induced by alcohol. Curr Top Behav Neurosci. 2013;13:31–86.

  54. 54.

    Soderpalm B, Lido HH, Ericson M. The glycine receptor-A functionally important primary brain target of ethanol. Alcohol Clin Exp Res. 2017;41:1816–30.

  55. 55.

    Cuzon Carlson VC, Grant KA, Lovinger DM. Synaptic adaptations to chronic ethanol intake in male rhesus monkey dorsal striatum depend on age of drinking onset. Neuropharmacology. 2018;131:128–42.

  56. 56.

    Mameli M, Zamudio PA, Carta M, Valenzuela CF. Developmentally regulated actions of alcohol on hippocampal glutamatergic transmission. J Neurosci. 2005;25:8027–36.

  57. 57.

    Zhu W, Bie B, Pan ZZ. Involvement of non-NMDA glutamate receptors in central amygdala in synaptic actions of ethanol and ethanol-induced reward behavior. J Neurosci. 2007;27:289–98.

  58. 58.

    Goodwani S, Saternos H, Alasmari F, Sari Y. Metabotropic and ionotropic glutamate receptors as potential targets for the treatment of alcohol use disorder. Neurosci Biobehav Rev. 2017;77:14–31.

  59. 59.

    Hopf FW. Do specific NMDA receptor subunits act as gateways for addictive behaviors? Genes Brain Behav. 2017;16:118–38.

  60. 60.

    Moghaddam B, Bolinao ML. Biphasic effect of ethanol on extracellular accumulation of glutamate in the hippocampus and the nucleus accumbens. Neurosci Lett. 1994;178:99–102.

  61. 61.

    Griffin WC 3rd, Haun HL, Hazelbaker CL, Ramachandra VS, Becker HC. Increased extracellular glutamate in the nucleus accumbens promotes excessive ethanol drinking in ethanol dependent mice. Neuropsychopharmacology. 2014;39:707–17.

  62. 62.

    Spanagel R. Alcoholism: a systems approach from molecular physiology to addictive behavior. Physiol Rev. 2009;89:649–705.

  63. 63.

    Renteria R, Buske TR, Morrisett RA. Long-term subregion-specific encoding of enhanced ethanol intake by D1DR medium spiny neurons of the nucleus accumbens. Addict Biol. 2018;23:689–98.

  64. 64.

    Besser L, Chorin E, Sekler I, Silverman WF, Atkin S, Russell JT, et al. Synaptically released zinc triggers metabotropic signaling via a zinc-sensing receptor in the hippocampus. J Neurosci. 2009;29:2890–901.

  65. 65.

    McKee KK, Tan CP, Palyha OC, Liu J, Feighner SD, Hreniuk DL, et al. Cloning and characterization of two human G protein-coupled receptor genes (GPR38 and GPR39) related to the growth hormone secretagogue and neurotensin receptors. Genomics. 1997;46:426–34.

  66. 66.

    Popovics P, Stewart AJ. GPR39: a Zn(2+)-activated G protein-coupled receptor that regulates pancreatic, gastrointestinal and neuronal functions. Cell Mol life Sci. 2011;68:85–95.

  67. 67.

    Frimurer TM, Mende F, Graae AS, Engelstoft MS, Egerod KL, Nygaard R, et al. Model-based discovery of synthetic agonists for the Zn(2+)-sensing G-protein-coupled receptor 39 (GPR39) reveals novel biological functions. J Med Chem. 2017;60:886–98.

  68. 68.

    Hashimoto H, Ueta Y. Central effects of ghrelin, a unique peptide, on appetite and fluid/water drinking behavior. Curr Protein Pept Sci. 2011;12:280–7.

  69. 69.

    Hashimoto H, Otsubo H, Fujihara H, Suzuki H, Ohbuchi T, Yokoyama T, et al. Centrally administered ghrelin potently inhibits water intake induced by angiotensin II and hypovolemia in rats. J Physiol Sci. 2010;60:19–25.

  70. 70.

    Farokhnia M, Grodin EN, Lee MR, Oot EN, Blackburn AN, Stangl BL, et al. Exogenous ghrelin administration increases alcohol self-administration and modulates brain functional activity in heavy-drinking alcohol-dependent individuals. Mol Psychiatry. 2017;23:2029–38.

  71. 71.

    Leggio L, Zywiak WH, Fricchione SR, Edwards SM, de la Monte SM, Swift RM, et al. Intravenous ghrelin administration increases alcohol craving in alcohol-dependent heavy drinkers: a preliminary investigation. Biol Psychiatry. 2014;76:734–41.

  72. 72.

    Zallar LJ, Farokhnia M, Tunstall BJ, Vendruscolo LF, Leggio L. The role of the ghrelin system in drug addiction. Int Rev Neurobiol. 2017;136:89–119.

  73. 73.

    Sunuwar L, Medini M, Cohen L, Sekler I, Hershfinkel M. The zinc sensing receptor, ZnR/GPR39, triggers metabotropic calcium signalling in colonocytes and regulates occludin recovery in experimental colitis. Philos Trans R Soc Lond B Biol Sci. 2016;371:1700.

  74. 74.

    Leclercq S, de Timary P, Delzenne NM, Starkel P. The link between inflammation, bugs, the intestine and the brain in alcohol dependence. Transl Psychiatry. 2017;7:e1048.

  75. 75.

    Kasarskis EJ, Manton WI, Devenport LD, Kirkpatrick JB, Howell GA, Klitenick MA, et al. Effects of alcohol ingestion on zinc content of human and rat central nervous systems. Exp Neurol. 1985;90:81–95.

  76. 76.

    Hu KH, Friede RL. Topographic determination of zinc in human brain by atomic absorption spectrophotometry. J Neurochem. 1968;15:677–85.

  77. 77.

    Boden JM, Fergusson DM. Alcohol and depression. Addiction. 2011;106:906–14.

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Acknowledgements

The authors thank Houda Mesnaoui for technical assistance.

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Affiliations

  1. Division of Neuroscience, Oregon National Primate Research Center, Oregon Health and Sciences University, Beaverton, Oregon, USA

    • Verginia C. Cuzon Carlson
    • , Matthew M. Ford
    • , Timothy L. Carlson
    • , Kathleen A. Grant
    • , Betsy Ferguson
    •  & Rita P. Cervera-Juanes
  2. Department of Behavioral Neuroscience, Oregon Health and Science University, Portland, Oregon, USA

    • Verginia C. Cuzon Carlson
    • , Matthew M. Ford
    •  & Kathleen A. Grant
  3. Division of Genetics, Oregon National Primate Research, Oregon Health and Sciences University, Beaverton, Oregon, USA

    • Alejandro Lomniczi
    • , Betsy Ferguson
    •  & Rita P. Cervera-Juanes
  4. Department of Molecular and Medical Genetics, Oregon Health and Sciences University, Portland, Oregon, USA

    • Betsy Ferguson

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Correspondence to Rita P. Cervera-Juanes.

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https://doi.org/10.1038/s41386-018-0308-1