Acta Diabetologica

, Volume 53, Issue 2, pp 279–293 | Cite as

GPR39 receptors and actions of trace metals on pancreatic beta cell function and glucose homoeostasis

  • Brian. M. Moran
  • Yasser H. A. Abdel-Wahab
  • Srividya Vasu
  • Peter R. Flatt
  • Aine M. McKillopEmail author
Original Article



G-protein-coupled receptor 39 (GPR39) has been implicated in glucose homoeostasis, appetite control and gastrointestinal tract function.


This study used clonal BRIN-BD11 cells and mouse pancreatic islets to assess the insulin-releasing actions of trace metals believed to act via GPR39, and the second messenger pathways involved in mediating their effects. Micromolar concentrations of Zn2+, Cu2+, Ni2+ and Co2+ were examined under normoglycaemic and hyperglycaemic conditions. Mechanistic studies investigated changes of intracellular Ca2+, cAMP generation and assessment of cytotoxicity by LDH release. Cellular localisation of GPR39 was determined by double immunohistochemical staining.


All trace metals (7.8–500 µmol/l) stimulated insulin release with Cu2+ being the most potent in isolated islets, with an EC50 value of 87 μmol/l. Zn2+ was the most selective with an EC50 value of 125 μmol/l. Enhancement of insulin secretion was also observed with Ni2+ (179 μmol/l) and Co2+ (190 μmol/l). These insulin-releasing effects were confirmed using clonal BRIN-BD11 cells which exhibited enhanced intracellular Ca2+ (p < 0.05–p < 0.001) and cAMP generation (p < 0.05–p < 0.001) in response to trace metals. Oral administration of Zn2+, Ni2+ and Cu2+ (50 µmol/kg together with 18 mmol/kg glucose) decreased the glycaemic excursion (p < 0.05–p < 0.01) and augmented insulin secretion (p < 0.05–p < 0.01) in NIH Swiss mice.


This study has demonstrated the presence of GPR39 and the insulinotropic actions of trace metals on BRIN-BD11 cells and pancreatic beta cells, together with their antihyperglycaemic actions in vivo. These data suggest that development of agonists capable of specifically activating GPR39 may be a useful new therapeutic approach for diabetes management.


G-protein-coupled receptor 39 Trace metals Pancreatic beta cells Glucose tolerance Insulin secretion 



These studies were supported by the Department of Education and Learning, Northern Ireland.

Conflict of interest

The authors declare that they have no conflict of interest.

Ethical standard

This study was approved by the University of Ulster Animal Ethics Review Committee. All animal experiments were carried out in accordance with the UK Animal (Scientific Procedures) Act 1986.

Human and animal rights

All procedures followed were in accordance with the UK Animal (Scientific Procedures) Act 1986 and the ARRIVE guidelines for reporting experiments involving animals. No clinical studies were carried out in this study.

Informed consent

No informed consent was required as no patients or clinical studies were involved in this study.


  1. 1.
    Tremblay F, Perreault M, Klaman LD, Tobin JE, Smith E, Gimeno RE (2007) Normal food intake and body weight in mice lacking the G protein-coupled receptor GPR39. Endocrinology 148:501–506CrossRefPubMedGoogle Scholar
  2. 2.
    Holst B, Holliday ND, Bach A, Elling CE, Cox HM, Schwartz TM (2004) Common structural basis for constitutive activity of the ghrelin receptor family. J Biol Chem 279:53806–53817CrossRefPubMedGoogle Scholar
  3. 3.
    Egerod KL, Holst B, Petersen PS, Hansen JB, Mulder J, Hokfelt T, Schwartz TW (2007) GPR39 splice variants versus antisense gene LYPD1: expression and regulation in gastrointestinal tract, endocrine pancreas, liver and white adipose tissue. Mol Endocrinol 21:1685–1698CrossRefPubMedGoogle Scholar
  4. 4.
    McKee KK, Tan CP, Palyha OC, Liu J, Feighner SD, Hreniuk DL, Smith RG, Howard AD, Van der Ploeg LH (1997) Cloning and characterization of two human G protein-coupled receptor genes (GPR38 and GPR39) related to growth hormone secretagogue and neurotensin receptors. Genomics 46:426–434CrossRefPubMedGoogle Scholar
  5. 5.
    Catalan V, Gomez-Ambrosi J, Rotellar F, Silva C, Gil MJ, Rodriquez A, Cienfuegos JA, Salvador J, Fruhbeck G (2007) The obestatin receptor (GPR39) is expressed in human adipose tissue and is down-regulated in obesity-associated type 2 diabetes mellitus. Clin Endocrinol 66:598–601Google Scholar
  6. 6.
    Egerod KL, Jin C, Petersen PS, Wierup N, Sundler F, Holst B, Schwartz TW (2011) β- cell specific overexpression of GPR39 protects against streptozotocin-induced hyperglycaemia. Int J Endocrinol 2011:1–8CrossRefGoogle Scholar
  7. 7.
    Holst B, Egerod KL, Jin C, Petersen PS, Ostergaard MV, Hald J, Sprinkel AM, Storling J, Mandrup-Poulsen T, Holst JJ et al (2009) G protein-coupled receptor 39 deficiency is associated with pancreatic islet dysfunction. Endocrinology 150:2577–2585CrossRefPubMedPubMedCentralGoogle Scholar
  8. 8.
    Xie F, Lui H, Zhu Y, Qin YR, Dai Y, Zeng T, Chen L, Nie C, Tang H, Li Y et al (2011) Overexpression of GPR39 contributes to malignant development of human esophageal squamous cell carcinoma. BMC Cancer 11:1–12CrossRefGoogle Scholar
  9. 9.
    Zhang JV, Ren P, Avsian-Kretchmer O, Luo CW, Rauch R, Klein C, Hsueh AJ (2005) Obestatin, a peptide encoded by the ghrelin gene, opposes ghrelin’s effects on food intake. Science 310:996–999CrossRefPubMedGoogle Scholar
  10. 10.
    Holst B, Egerod KL, Schild E, Vickers SP, Cheetham S, Gerlach LO, Storjohann L, Stidsen CE, Jones R, Beck-Sickinger AG et al (2007) GPR39 signaling is stimulated by zinc ions but not by obestatin. Endocrinology 148:13–20CrossRefPubMedGoogle Scholar
  11. 11.
    Gourcerol G, St-Pierre DH, Tache Y (2007) Lack of obestatin effects on food intake: should obestatin be renamed ghrelin-associated peptide (GAP)? Regul Pept 141:1–7CrossRefPubMedGoogle Scholar
  12. 12.
    Kobelt P, Wisser AS, Stengel A, Goebel M, Bannert N, Gourcerol G, Inhoff T, Noetzel S, Wiedenmann B, Klapp BF et al (2008) Peripheral obestatin has no effects on feeding behaviour and brain Fos expression in rodents. Peptides 29:1018–1027CrossRefPubMedPubMedCentralGoogle Scholar
  13. 13.
    Andersson T, Berggren PO, Flatt PR (1980) Subcellular distribution of zinc in islet β-cell fractions. Horm Metab Res 12:275–276CrossRefPubMedGoogle Scholar
  14. 14.
    Petersen PS, Jin C, Madsen AN, Rasmussen M, Kuhre R, Egerod KL, Nielsen LB, Schwartz TW, Holst B (2011) Deficiency of the GPR39 receptor is associated with obesity and altered adipocyte metabolism. FASEB J 25:3803–3814CrossRefPubMedGoogle Scholar
  15. 15.
    Kelleher SL, McCormick NH, Velasquez V, Lopez V (2011) Zinc in specialized secretory tissues: roles in the pancreas, prostate, and mammary gland. Adv Nutr 2:101–111CrossRefPubMedPubMedCentralGoogle Scholar
  16. 16.
    Popovics P, Stewart AJ (2011) GPR39: a Zn2+ -activated G-protein coupled receptor that regulates pancreatic, gastrointestinal and neuronal functions. Cell Mol Life Sci 68:85–95CrossRefPubMedGoogle Scholar
  17. 17.
    Holst B, Elling CE, Schwartz TW (2002) Metal ion-mediated agonism and agonist enhancement in melanocortin MC1 and MC4 receptors. J Biol Chem 277:47662–47670CrossRefPubMedGoogle Scholar
  18. 18.
    Ohly P, Dohle C, Abel J, Seissler J, Gleichmann H (2000) Zinc sulphate induces metallothionein in pancreatic islets of mice and protects against diabetes induced by multiple low doses of streptozotocin. Diabetologia 43:1020–1030CrossRefPubMedGoogle Scholar
  19. 19.
    Huber AM, Gershoff SN (1973) Effect of zinc deficiency in rats on insulin release from the pancreas. J Nutr 103:1739–1744PubMedGoogle Scholar
  20. 20.
    Chen MD, Liou SJ, Lin PY, Yang VC, Alexander PS, Lin WH (1998) Effects of zinc supplementation on the plasma glucose level and insulin activity in genetically obese (ob/ob) mice. Bio Trace Elem Res 61:303–311CrossRefGoogle Scholar
  21. 21.
    Depoortere I (2012) GI functions of GPR39: novel biology. Curr Opin Pharmacol 12:647–652CrossRefPubMedGoogle Scholar
  22. 22.
    Moechars D, Depoortere I, Moreaux B, de Smet B, Goris I, Hoskens L, Daneels G, Kass S, Ver Donck L, Peeters T et al (2006) Altered gastrointestinal and metabolic function in the GPR39-obestatin receptor knockout mouse. Gastroenterology 131:1131–1141CrossRefPubMedGoogle Scholar
  23. 23.
    Gault VA, Porter WD, Flatt PR, Holscher C (2010) Actions of exendin-4 therapy on cognitive function and hippocampal synaptic plasticity in mice fed a high fat diet. In J Obes 34:1341–1344CrossRefGoogle Scholar
  24. 24.
    Porter DW, Kerr BD, Flatt PR, Holscher C, Gault VA (2010) Four weeks administration of Liraglutide improves memory and learning as well as glycaemic control in mice with dietary-induced obesity and insulin resistance. Diabetes Obes Metab 12:891–899CrossRefPubMedGoogle Scholar
  25. 25.
    Besser L, Chorin E, Sekler I, Silverman WF, Atkin S, Russell J, Hershfinkel M (2009) Synaptically released zinc triggers metabotropic signaling via a zinc-sensing receptor in the hippocampus. J Neurosci 29:2890–2901CrossRefPubMedPubMedCentralGoogle Scholar
  26. 26.
    Dittmer S, Sahin M, Pantlen A, Saxena A, Toutzaris D, Pina AL, Geerts A, Golz S, Methner A (2008) 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 283:7074–7081CrossRefPubMedGoogle Scholar
  27. 27.
    Mlyneic K, Budziszewska B, Reczynski W, Sowa-Kucma M, Nowak G (2013) The role of the GPR39 receptor in zinc deficient-animal model of depression. Behav Brain Res 238:30–35CrossRefGoogle Scholar
  28. 28.
    Cole TB, Robbins CA, Wenzel HJ, Schwartzkroin PA, Palmiter RD (2000) Seizures and neuronal damage in mice lacking vesicular zinc. Epilepsy Res 39:153–169CrossRefPubMedGoogle Scholar
  29. 29.
    Sun SW, Won SJ, Hamby AM, Yoo BH, Fan Y, Sheline CT, Tamano H, Takeda A, Liu J (2009) Decreased brain zinc availability reduced hippocampal neurogenesis in mice and rats. J Cereb Blood Flow Metab 29:1579–1588CrossRefGoogle Scholar
  30. 30.
    Sharir H, Zinger A, Nevo N, Sekler I, Hershfinkel M (2010) Zinc released from injured cells is acting via the Zn2+-sensing receptor, ZnR, to trigger signaling leading to epithelial repair. J Biol Chem 285:26097–26106CrossRefPubMedPubMedCentralGoogle Scholar
  31. 31.
    McClenaghan NH, Barnett CR, Ah-Sing E, Abdel-Wahab YH, O’Harte FP, Yoon TW, Swanston-Flatt SK, Flatt PR (1996) Characterisation of a novel glucose-responsive insulin-secreting cell line, BRIN BD11, produced by electrofusion. Diabetes 45:1132–1140CrossRefPubMedGoogle Scholar
  32. 32.
    Flatt PR, Bailey CJ (1981) Abnormal plasma glucose and insulin responses in heterozygous lean (ob/+) mice. Diabetologia 20:573–577CrossRefPubMedGoogle Scholar
  33. 33.
    Moskalewski S (1969) Studies on the culture and transplantation of isolated islets of Langerhans of the guinea pig. Proc K Ned Akad Wet C 72:157–171PubMedGoogle Scholar
  34. 34.
    Bailey CJ, Flatt PR (1982) Influence of genetic background and age on the expression of the obese hyperglycaemic syndrome in Aston ob/ob mice. Int J Obes 6:11–21PubMedGoogle Scholar
  35. 35.
    Hannan JMA, Marenah L, Ali L, Rokeya B, Flatt PR, Abdel-Wahab YHA (2006) Ocimum sanctum leaf extracts stimulate insulin secretion from perfused pancreas, isolated islets and clonal pancreatic β-cells. J Endocrinol 189:127–136CrossRefPubMedGoogle Scholar
  36. 36.
    Miguel JC, Patterson S, Abdel-Wahab YHA, Mathias PC, Flatt PR (2004) Time-correlation between membrane depolarization and intracellular calcium in insulin secretion BRIN-BD11 cells: studies using FLIPR. Cell Calcium 36:43–50CrossRefPubMedGoogle Scholar
  37. 37.
    Moran BM, Abdel-Wahab YH, Flatt PR, McKillop AM (2014) Evaluation of the insulin releasing and glucose lowering effects of GPR120 activation in pancreatic beta cells. Diabetes Obes Metab 16:1128–1139CrossRefPubMedGoogle Scholar
  38. 38.
    Kilkenny C, Browne W, Cuthill IC, Emerson M, Altman DG (2010) Animal research: reporting in vivo experiments: the ARRIVE guidelines. Br J Pharmacol 160:1577–1579CrossRefPubMedPubMedCentralGoogle Scholar
  39. 39.
    Vangaveti V, Shashidhar V, Jarrod G, Baune BT, Kennedy RL (2010) Free fatty acid receptors: emerging targets for treatment of diabetes and its complications. Ther Adv Endocrinol Metab 1:165–175CrossRefPubMedPubMedCentralGoogle Scholar
  40. 40.
    Bailey CJ (2005) Drugs on the horizon for diabesity. Curr Diabet Rep 5:353–359CrossRefGoogle Scholar
  41. 41.
    Burant CF (2013) Activation of GPR40 as a therapeutic target for the treatment of type 2 diabetes. Diabetes Care 36:S175–S179CrossRefPubMedPubMedCentralGoogle Scholar
  42. 42.
    Cohen L, Azriel-Tamir H, Arotsker N, Sekler I, Hershfinkel M (2012) Zinc sensing receptor signalling, mediated by GPR39, reduces butyrate-induced cell death in HT29 colonocytes via upregulation of clusterin. PloS One 7:e35482CrossRefPubMedPubMedCentralGoogle Scholar
  43. 43.
    Tremblay F, Richard A, Will S, Syed J, Stedman N, Perreault M, Gimeno RE (2009) Distribution of G-protein-coupled receptor 39 impairs insulin secretion in vivo. Endocrinology 150:2586–2595CrossRefPubMedGoogle Scholar
  44. 44.
    Rutter GA (2010) Think zinc: new roles for zinc in the control of insulin secretion. Islets 2:49–50CrossRefPubMedGoogle Scholar
  45. 45.
    Lodemann U, Einspanier R, Scharfen F, Martens H, Bondzio A (2013) Effects of zinc on epithelial barrier properties and viability in a human and a porcine intestinal cell culture model. Toxicol In Vitro 27:834–843CrossRefPubMedGoogle Scholar
  46. 46.
    Flatt PR, Rorsman P, Swanston-Flatt SK (1987) Effect of cationic modifications on superficial binding and intracellular 45Ca uptake by decapsulated ob/ob mouse pancreatic islets. Biomed Res 8:153–159Google Scholar
  47. 47.
    Bloc A, Cens T, Cruz H, Dunant Y (2000) Zinc-induced changes in ionic currents of clonal rat pancreatic β-cells: activation of ATP-sensitive K+ channels. J Physiol 529:723–734CrossRefPubMedPubMedCentralGoogle Scholar
  48. 48.
    Acuna-Castillo C, Morales B, Huidobro-Toro JP (2000) Zinc and copper modulate differentially the P2X4 receptor. J Neurosci 74:1529–1537Google Scholar
  49. 49.
    Fraga CG (2005) Relevance, essentiality and toxicity of trace elements in human health. Mol Asp Med 26:235–244CrossRefGoogle Scholar
  50. 50.
    Tanaka T, Yano T, Adachi T, Koshimizu TA, Hirasawa A, Tsujimoto G (2008) Cloning and characterization of the rat free fatty acid receptor GPR120: in vivo effect of the natural ligand on GLP-1 secretion and proliferation of pancreatic beta cells. N-S Arch Pharmacol 377:515–522CrossRefGoogle Scholar
  51. 51.
    Lauffer LM, Iakoubov R, Brubaker PL (2009) GPR119 is essential for oleoylethanolamide–induced glucagon-like peptide-1 secretion from the intestinal enteroendocrine L-cells. Diabetes 58:1058–1066CrossRefPubMedPubMedCentralGoogle Scholar
  52. 52.
    Adachi Y, Yoshida J, Kodera Y, Kiss T, Jakusch T, Enyedy EA, Yoshikawa Y, Sakurai H (2006) Oral administration of a zinc complex improves type 2 diabetes and metabolic syndromes. Biochem Biophys Res Commun 351:165–170CrossRefPubMedGoogle Scholar

Copyright information

© Springer-Verlag Italia 2015

Authors and Affiliations

  • Brian. M. Moran
    • 1
  • Yasser H. A. Abdel-Wahab
    • 1
  • Srividya Vasu
    • 1
  • Peter R. Flatt
    • 1
  • Aine M. McKillop
    • 1
    Email author
  1. 1.Biomedical Sciences Research Institute, SAAD Centre for Pharmacy and DiabetesUniversity of UlsterColeraineNorthern Ireland, UK

Personalised recommendations