Skip to main content
PMC home page
The EMBO Journal logoLink to The EMBO Journal
. 1988 Aug;7(8):2443–2449. doi: 10.1002/j.1460-2075.1988.tb03090.x

Activators of protein kinase C depolarize insulin-secreting cells by closing K+ channels.

C B Wollheim 1, M J Dunne 1, B Peter-Riesch 1, R Bruzzone 1, T Pozzan 1, O H Petersen 1
PMCID: PMC457113  PMID: 3056715

Abstract

Carbohydrate stimuli of insulin secretion depolarize the pancreatic B cell and the B-cell line RINm5F by inhibiting ATP-sensitive K+ channels. We examined the possibility that this effect is mediated by activation of protein kinase C. In RINm5F cells, the triose D-glyceraldehyde evoked a rapid increase of the mass of 1,2-diacylglycerol, the endogenous activator of protein kinase C. This effect is mainly due to de novo synthesis of the lipid from glycolytic intermediates, as glyceraldehyde carbon was incorporated into 1,2-diacylglycerol within 1 min of exposure to 14C-labelled glyceraldehyde. The effects of two exogenous activators of kinase C, 4-beta-12-phorbol-myristate 13-acetate (PMA) and 1,2-didecanoylglycerol (DC10) on single K+ channel currents were examined in RINm5F cell-attached membrane patches. Both PMA and DC10 depolarized the cells and decreased the open-state probability of the ATP-sensitive K+ channels. These actions were not due to changes in cellular ATP content, since PMA, like glyceraldehyde, failed to alter cellular ATP. As is the case for glyceraldehyde, PMA and DC10 raised cytosolic free Ca2+ [( Ca2+]i) and stimulated insulin secretion. Both of these effects are inhibited in the absence of external Ca2+. This, and the attenuation of the [Ca2+]i rise by verapamil, suggest that all three stimuli raise [Ca2+]i by promoting Ca2+ influx through voltage-gated channels in turn leading to insulin secretion. As the exogenous activators of protein kinase C mimic the effects of glyceraldehyde, it is proposed that the carbohydrate-mediated production of 1,2-diacylglycerol constitutes the link between metabolism and membrane depolarization.(ABSTRACT TRUNCATED AT 250 WORDS)

Full text

PDF
2443

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Arkhammar P., Nilsson T., Rorsman P., Berggren P. O. Inhibition of ATP-regulated K+ channels precedes depolarization-induced increase in cytoplasmic free Ca2+ concentration in pancreatic beta-cells. J Biol Chem. 1987 Apr 25;262(12):5448–5454. [PubMed] [Google Scholar]
  2. Ashcroft F. M., Ashcroft S. J., Harrison D. E. Effects of 2-ketoisocaproate on insulin release and single potassium channel activity in dispersed rat pancreatic beta-cells. J Physiol. 1987 Apr;385:517–529. doi: 10.1113/jphysiol.1987.sp016505. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Ashcroft F. M., Harrison D. E., Ashcroft S. J. Glucose induces closure of single potassium channels in isolated rat pancreatic beta-cells. 1984 Nov 29-Dec 5Nature. 312(5993):446–448. doi: 10.1038/312446a0. [DOI] [PubMed] [Google Scholar]
  4. Ashcroft S. J., Weerasinghe L. C., Randle P. J. Interrelationship of islet metabolism, adenosine triphosphate content and insulin release. Biochem J. 1973 Feb;132(2):223–231. doi: 10.1042/bj1320223. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Banschbach M. W., Geison R. L., Hokin-Neaverson M. Effects of cholinergic stimulation on levels and fatty acid composition of diacylglycerols in mouse pancreas. Biochim Biophys Acta. 1981 Jan 26;663(1):34–45. doi: 10.1016/0005-2760(81)90192-2. [DOI] [PubMed] [Google Scholar]
  6. Biden T. J., Peter-Riesch B., Schlegel W., Wollheim C. B. Ca2+-mediated generation of inositol 1,4,5-triphosphate and inositol 1,3,4,5-tetrakisphosphate in pancreatic islets. Studies with K+, glucose, and carbamylcholine. J Biol Chem. 1987 Mar 15;262(8):3567–3571. [PubMed] [Google Scholar]
  7. Bozem M., Nenquin M., Henquin J. C. The ionic, electrical, and secretory effects of protein kinase C activation in mouse pancreatic B-cells: studies with a phorbol ester. Endocrinology. 1987 Sep;121(3):1025–1033. doi: 10.1210/endo-121-3-1025. [DOI] [PubMed] [Google Scholar]
  8. Cockcroft S., Allan D. The fatty acid composition of phosphatidylinositol, phosphatidate and 1,2-diacylglycerol in stimulated human neutrophils. Biochem J. 1984 Sep 1;222(2):557–559. doi: 10.1042/bj2220557. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Cook D. L., Hales C. N. Intracellular ATP directly blocks K+ channels in pancreatic B-cells. Nature. 1984 Sep 20;311(5983):271–273. doi: 10.1038/311271a0. [DOI] [PubMed] [Google Scholar]
  10. Di Virgilio F., Pozzan T., Wollheim C. B., Vicentini L. M., Meldolesi J. Tumor promoter phorbol myristate acetate inhibits Ca2+ influx through voltage-gated Ca2+ channels in two secretory cell lines, PC12 and RINm5F. J Biol Chem. 1986 Jan 5;261(1):32–35. [PubMed] [Google Scholar]
  11. Dunlop M. E., Larkins R. G. Pancreatic islets synthesize phospholipids de novo from glucose via acyl-dihydroxyacetone phosphate. Biochem Biophys Res Commun. 1985 Oct 30;132(2):467–473. doi: 10.1016/0006-291x(85)91157-x. [DOI] [PubMed] [Google Scholar]
  12. Dunne M. J., Findlay I., Petersen O. H. Effects of pyridine nucleotides on the gating of ATP-sensitive potassium channels in insulin-secreting cells. J Membr Biol. 1988 Jun;102(3):205–216. doi: 10.1007/BF01925714. [DOI] [PubMed] [Google Scholar]
  13. Dunne M. J., Findlay I., Petersen O. H., Wollheim C. B. ATP-sensitive K+ channels in an insulin-secreting cell line are inhibited by D-glyceraldehyde and activated by membrane permeabilization. J Membr Biol. 1986;93(3):271–279. doi: 10.1007/BF01871181. [DOI] [PubMed] [Google Scholar]
  14. Dunne M. J., Petersen O. H. Intracellular ADP activates K+ channels that are inhibited by ATP in an insulin-secreting cell line. FEBS Lett. 1986 Nov 10;208(1):59–62. doi: 10.1016/0014-5793(86)81532-0. [DOI] [PubMed] [Google Scholar]
  15. Farese R. V., DiMarco P. E., Barnes D. E., Sabir M. A., Larson R. E., Davis J. S., Morrison A. D. Rapid glucose-dependent increases in phosphatidic acid and phosphoinositides in rat pancreatic islets. Endocrinology. 1986 Apr;118(4):1498–1503. doi: 10.1210/endo-118-4-1498. [DOI] [PubMed] [Google Scholar]
  16. Findlay I., Dunne M. J., Petersen O. H. ATP-sensitive inward rectifier and voltage- and calcium-activated K+ channels in cultured pancreatic islet cells. J Membr Biol. 1985;88(2):165–172. doi: 10.1007/BF01868430. [DOI] [PubMed] [Google Scholar]
  17. Findlay I., Dunne M. J., Petersen O. H. High-conductance K+ channel in pancreatic islet cells can be activated and inactivated by internal calcium. J Membr Biol. 1985;83(1-2):169–175. doi: 10.1007/BF01868748. [DOI] [PubMed] [Google Scholar]
  18. Findlay I., Dunne M. J., Ullrich S., Wollheim C. B., Petersen O. H. Quinine inhibits Ca2+-independent K+ channels whereas tetraethylammonium inhibits Ca2+-activated K+ channels in insulin-secreting cells. FEBS Lett. 1985 Jun 3;185(1):4–8. doi: 10.1016/0014-5793(85)80729-8. [DOI] [PubMed] [Google Scholar]
  19. Findlay I., Dunne M. J. Voltage-activated Ca2+ currents in insulin-secreting cells. FEBS Lett. 1985 Sep 23;189(2):281–285. doi: 10.1016/0014-5793(85)81040-1. [DOI] [PubMed] [Google Scholar]
  20. Grynkiewicz G., Poenie M., Tsien R. Y. A new generation of Ca2+ indicators with greatly improved fluorescence properties. J Biol Chem. 1985 Mar 25;260(6):3440–3450. [PubMed] [Google Scholar]
  21. Hamill O. P., Marty A., Neher E., Sakmann B., Sigworth F. J. Improved patch-clamp techniques for high-resolution current recording from cells and cell-free membrane patches. Pflugers Arch. 1981 Aug;391(2):85–100. doi: 10.1007/BF00656997. [DOI] [PubMed] [Google Scholar]
  22. Henquin J. C., Meissner H. P. Significance of ionic fluxes and changes in membrane potential for stimulus-secretion coupling in pancreatic B-cells. Experientia. 1984 Oct 15;40(10):1043–1052. doi: 10.1007/BF01971450. [DOI] [PubMed] [Google Scholar]
  23. Higashida H., Brown D. A. Two polyphosphatidylinositide metabolites control two K+ currents in a neuronal cell. 1986 Sep 25-Oct 1Nature. 323(6086):333–335. doi: 10.1038/323333a0. [DOI] [PubMed] [Google Scholar]
  24. Kakei M., Kelly R. P., Ashcroft S. J., Ashcroft F. M. The ATP-sensitivity of K+ channels in rat pancreatic B-cells is modulated by ADP. FEBS Lett. 1986 Nov 10;208(1):63–66. doi: 10.1016/0014-5793(86)81533-2. [DOI] [PubMed] [Google Scholar]
  25. Lapetina E. G., Reep B., Ganong B. R., Bell R. M. Exogenous sn-1,2-diacylglycerols containing saturated fatty acids function as bioregulators of protein kinase C in human platelets. J Biol Chem. 1985 Feb 10;260(3):1358–1361. [PubMed] [Google Scholar]
  26. Lernmark A. Molecular biology of type 1 (insulin-dependent) diabetes mellitus. Diabetologia. 1985 Apr;28(4):195–203. doi: 10.1007/BF00282232. [DOI] [PubMed] [Google Scholar]
  27. Malaisse W. J., Dunlop M. E., Mathias P. C., Malaisse-Lagae F., Sener A. Stimulation of protein kinase C and insulin release by 1-oleoyl-2-acetyl-glycerol. Eur J Biochem. 1985 May 15;149(1):23–27. doi: 10.1111/j.1432-1033.1985.tb08887.x. [DOI] [PubMed] [Google Scholar]
  28. Malaisse W. J., Hutton J. C., Kawazu S., Herchuelz A., Valverde I., Sener A. The stimulus-secretion coupling of glucose-induced insulin release. XXXV. The links between metabolic and cationic events. Diabetologia. 1979 May;16(5):331–341. doi: 10.1007/BF01223623. [DOI] [PubMed] [Google Scholar]
  29. Malaisse W. J., Lebrun P., Herchuelz A., Sener A., Malaisse-Lagae F. Synergistic effect of a tumor-promoting phorbol ester and a hypoglycemic sulfonylurea upon insulin release. Endocrinology. 1983 Nov;113(5):1870–1877. doi: 10.1210/endo-113-5-1870. [DOI] [PubMed] [Google Scholar]
  30. Malaisse W. J., Sener A. Glucose-induced changes in cytosolic ATP content in pancreatic islets. Biochim Biophys Acta. 1987 Feb 18;927(2):190–195. doi: 10.1016/0167-4889(87)90134-0. [DOI] [PubMed] [Google Scholar]
  31. Misler S., Falke L. C., Gillis K., McDaniel M. L. A metabolite-regulated potassium channel in rat pancreatic B cells. Proc Natl Acad Sci U S A. 1986 Sep;83(18):7119–7123. doi: 10.1073/pnas.83.18.7119. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Nishizuka Y. Studies and perspectives of protein kinase C. Science. 1986 Jul 18;233(4761):305–312. doi: 10.1126/science.3014651. [DOI] [PubMed] [Google Scholar]
  33. Pace C. S., Goldsmith K. T. Action of a phorbol ester on B-cells: potentiation of stimulant-induced electrical activity. Am J Physiol. 1985 May;248(5 Pt 1):C527–C534. doi: 10.1152/ajpcell.1985.248.5.C527. [DOI] [PubMed] [Google Scholar]
  34. Peter-Riesch B., Fathi M., Schlegel W., Wollheim C. B. Glucose and carbachol generate 1,2-diacylglycerols by different mechanisms in pancreatic islets. J Clin Invest. 1988 Apr;81(4):1154–1161. doi: 10.1172/JCI113430. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Petersen O. H., Findlay I. Electrophysiology of the pancreas. Physiol Rev. 1987 Jul;67(3):1054–1116. doi: 10.1152/physrev.1987.67.3.1054. [DOI] [PubMed] [Google Scholar]
  36. Prentki M., Wollheim C. B. Cytosolic free Ca2+ in insulin secreting cells and its regulation by isolated organelles. Experientia. 1984 Oct 15;40(10):1052–1060. doi: 10.1007/BF01971451. [DOI] [PubMed] [Google Scholar]
  37. Rorsman P., Trube G. Glucose dependent K+-channels in pancreatic beta-cells are regulated by intracellular ATP. Pflugers Arch. 1985 Dec;405(4):305–309. doi: 10.1007/BF00595682. [DOI] [PubMed] [Google Scholar]
  38. Trautmann M. E., Wollheim C. B. Characterization of glucose transport in an insulin-secreting cell line. Biochem J. 1987 Mar 15;242(3):625–630. doi: 10.1042/bj2420625. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Vallar L., Biden T. J., Wollheim C. B. Guanine nucleotides induce Ca2+-independent insulin secretion from permeabilized RINm5F cells. J Biol Chem. 1987 Apr 15;262(11):5049–5056. [PubMed] [Google Scholar]
  40. Vara E., Tamarit-Rodriguez J. Glucose stimulation of insulin secretion in islets of fed and starved rats and its dependence on lipid metabolism. Metabolism. 1986 Mar;35(3):266–271. doi: 10.1016/0026-0495(86)90212-x. [DOI] [PubMed] [Google Scholar]
  41. Velasco J. M., Petersen J. U., Petersen O. H. Single-channel Ba2+ currents in insulin-secreting cells are activated by glyceraldehyde stimulation. FEBS Lett. 1988 Apr 25;231(2):366–370. doi: 10.1016/0014-5793(88)80851-2. [DOI] [PubMed] [Google Scholar]
  42. Velasco J. M., Petersen O. H. Voltage-activation of high-conductance K+ channel in the insulin-secreting cell line RINm5F is dependent on local extracellular Ca2+ concentration. Biochim Biophys Acta. 1987 Jan 26;896(2):305–310. doi: 10.1016/0005-2736(87)90191-x. [DOI] [PubMed] [Google Scholar]
  43. Virji M. A., Steffes M. W., Estensen R. D. Phorbol myristate acetate: effect of a tumor promoter on insulin release from isolated rat islets of Langerhans. Endocrinology. 1978 Mar;102(3):706–711. doi: 10.1210/endo-102-3-706. [DOI] [PubMed] [Google Scholar]
  44. Weiss J. N., Lamp S. T. Glycolysis preferentially inhibits ATP-sensitive K+ channels in isolated guinea pig cardiac myocytes. Science. 1987 Oct 2;238(4823):67–69. doi: 10.1126/science.2443972. [DOI] [PubMed] [Google Scholar]
  45. Wollheim C. B., Biden T. J. Second messenger function of inositol 1,4,5-trisphosphate. Early changes in inositol phosphates, cytosolic Ca2+, and insulin release in carbamylcholine-stimulated RINm5F cells. J Biol Chem. 1986 Jun 25;261(18):8314–8319. [PubMed] [Google Scholar]
  46. Wollheim C. B., Biden T. J. Signal transduction in insulin secretion: comparison between fuel stimuli and receptor agonists. Ann N Y Acad Sci. 1986;488:317–333. doi: 10.1111/j.1749-6632.1986.tb46568.x. [DOI] [PubMed] [Google Scholar]
  47. Wollheim C. B., Pozzan T. Correlation between cytosolic free Ca2+ and insulin release in an insulin-secreting cell line. J Biol Chem. 1984 Feb 25;259(4):2262–2267. [PubMed] [Google Scholar]
  48. Wollheim C. B., Ullrich S., Pozzan T. Glyceraldehyde, but not cyclic AMP-stimulated insulin release is preceded by a rise in cytosolic free Ca2+. FEBS Lett. 1984 Nov 5;177(1):17–22. doi: 10.1016/0014-5793(84)80972-2. [DOI] [PubMed] [Google Scholar]
  49. de Weille J., Schmid-Antomarchi H., Fosset M., Lazdunski M. ATP-sensitive K+ channels that are blocked by hypoglycemia-inducing sulfonylureas in insulin-secreting cells are activated by galanin, a hyperglycemia-inducing hormone. Proc Natl Acad Sci U S A. 1988 Feb;85(4):1312–1316. doi: 10.1073/pnas.85.4.1312. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from The EMBO Journal are provided here courtesy of Nature Publishing Group

RESOURCES