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Diabetologia

Erschienen in:

01.02.2004 | Article

Insulin secretion profiles are modified by overexpression of glutamate dehydrogenase in pancreatic islets

verfasst von: S. Carobbio, H. Ishihara, S. Fernandez-Pascual, C. Bartley, R. Martin-Del-Rio, P. Maechler

Erschienen in: Diabetologia | Ausgabe 2/2004

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Abstract

Aims/hypothesis

Glutamate dehydrogenase (GDH) is a mitochondrial enzyme playing a key role in the control of insulin secretion. However, it is not known whether GDH expression levels in beta cells are rate-limiting for the secretory response to glucose. GDH also controls glutamine and glutamate oxidative metabolism, which is only weak in islets if GDH is not allosterically activated by L-leucine or (+/−)-2-aminobicyclo-[2,2,1]heptane-2-carboxylic acid (BCH).

Methods

We constructed an adenovirus encoding for GDH to overexpress the enzyme in the beta-cell line INS-1E, as well as in isolated rat and mouse pancreatic islets. The secretory responses to glucose and glutamine were studied in static and perifusion experiments. Amino acid concentrations and metabolic parameters were measured in parallel.

Results

GDH overexpression in rat islets did not change insulin release at basal or intermediate glucose (2.8 and 8.3 mmol/l respectively), but potentiated the secretory response at high glucose concentrations (16.7 mmol/l) compared to controls (+35%). Control islets exposed to 5 mmol/l glutamine at basal glucose did not increase insulin release, unless BCH was added with a resulting 2.5-fold response. In islets overexpressing GDH glutamine alone stimulated insulin secretion (2.7-fold), which was potentiated 2.2-fold by adding BCH. The secretory responses evoked by glutamine under these conditions correlated with enhanced cellular metabolism.

Conclusions/interpretation

GDH could be rate-limiting in glucose-induced insulin secretion, as GDH overexpression enhanced secretory responses. Moreover, GDH overexpression made islets responsive to glutamine, indicating that under physiological conditions this enzyme acts as a gatekeeper to prevent amino acids from being inappropriate efficient secretagogues.

Michaelidis TM, Tzimagiorgis G, Moschonas NK, Papamatheakis J (1993) The human glutamate dehydrogenase gene family: gene organization and structural characterization. Genomics 16:150–160CrossRefPubMed

Hudson RC, Daniel RM (1993) L-glutamate dehydrogenases: distribution, properties and mechanism. Comp Biochem Physiol B 106:767–792CrossRefPubMed

Anderson CM, Swanson RA (2000) Astrocyte glutamate transport: review of properties, regulation, and physiological functions. Glia 32:1–14CrossRefPubMed

Nissim I (1999) Newer aspects of glutamine/glutamate metabolism: the role of acute pH changes. Am J Physiol 277:F493–F497PubMed

Sener A, Malaisse WJ (1980) L-leucine and a nonmetabolized analogue activate pancreatic islet glutamate dehydrogenase. Nature 288:187–189PubMed

Bryla J, Michalik M, Nelson J, Erecinska M (1994) Regulation of the glutamate dehydrogenase activity in rat islets of Langerhans and its consequence on insulin release. Metabolism 43:1187–1195PubMed

Yang SJ, Huh JW, Kim MJ et al. (2003) Regulatory effects of 5’-deoxypyridoxal on glutamate dehydrogenase activity and insulin secretion in pancreatic islets. Biochimie 85:581–586CrossRefPubMed

Stanley CA, Lieu YK, Hsu BY et al. (1998) Hyperinsulinism and hyperammonemia in infants with regulatory mutations of the glutamate dehydrogenase gene. N Engl J Med 338:1352–1357

Yorifuji T, Muroi J, Uematsu A, Hiramatsu H, Momoi T (1999) Hyperinsulinism-hyperammonemia syndrome caused by mutant glutamate dehydrogenase accompanied by novel enzyme kinetics. Hum Genet 104:476–479

10.

Fisher HF (1985) L-Glutamate dehydrogenase from bovine liver. Methods Enzymol 113:16–27PubMed

11.

Smith TJ, Peterson PE, Schmidt T, Fang J, Stanley CA (2001) Structures of bovine glutamate dehydrogenase complexes elucidate the mechanism of purine regulation. J Mol Biol 307:707–720CrossRefPubMed

12.

Stanley CA, Fang J, Kutyna K et al. (2000) Molecular basis and characterization of the hyperinsulinism/hyperammonemia syndrome: predominance of mutations in exons 11 and 12 of the glutamate dehydrogenase gene. HI/HA Contributing Investigators. Diabetes 49:667–673PubMed

13.

Maechler P, Wollheim CB (1999) Mitochondrial glutamate acts as a messenger in glucose-induced insulin exocytosis. Nature 402:685–689PubMed

14.

MacDonald MJ, Fahien LA (2000) Glutamate is not a messenger in insulin secretion. J Biol Chem 275:34025–34027CrossRefPubMed

15.

Rubi B, Ishihara H, Hegardt FG, Wollheim CB, Maechler P (2001) GAD65-mediated glutamate decarboxylation reduces glucose-stimulated insulin secretion in pancreatic beta cells. J Biol Chem 276:36391–36396CrossRefPubMed

16.

Bertrand G, Ishiyama N, Nenquin M, Ravier MA, Henquin JC (2002) The elevation of glutamate content and the amplification of insulin secretion in glucose-stimulated pancreatic islets are not causally related. J Biol Chem 277:32883–32891CrossRefPubMed

17.

Hoy M, Maechler P, Efanov AM, Wollheim CB, Berggren PO, Gromada J (2002) Increase in cellular glutamate levels stimulates exocytosis in pancreatic beta-cells. FEBS Lett 531:199–203CrossRefPubMed

18.

Liu YJ, Cheng H, Drought H, MacDonald MJ, Sharp GW, Straub SG (2003) Activation of the KATP channel-independent signaling pathway by the nonhydrolyzable analog of leucine, BCH. Am J Physiol Endocrinol Metab 285:E380–E389PubMed

19.

Henquin JC (2000) Triggering and amplifying pathways of regulation of insulin secretion by glucose. Diabetes 49:1751–1760PubMed

20.

Maechler P, Wollheim CB (2001) Mitochondrial function in normal and diabetic beta-cells. Nature 414:807–812PubMed

21.

Miki T, Nagashima K, Seino S (1999) The structure and function of the ATP-sensitive K+ channel in insulin-secreting pancreatic beta-cells. J Mol Endocrinol 22:113–123PubMed

22.

Rorsman P (1997) The pancreatic beta-cell as a fuel sensor: an electrophysiologist’s viewpoint. Diabetologia 40:487–495PubMed

23.

Lang J (1999) Molecular mechanisms and regulation of insulin exocytosis as a paradigm of endocrine secretion. Eur J Biochem 259:3–17CrossRefPubMed

24.

Malaisse WJ, Sener A, Carpinelli AR et al. (1980) The stimulus-secretion coupling of glucose-induced insulin release. XLVI. Physiological role of L-glutamine as a fuel for pancreatic islets. Mol Cell Endocrinol 20:171–189CrossRefPubMed

25.

Panten U, Zielmann S, Langer J, Zunkler BJ, Lenzen S (1984) Regulation of insulin secretion by energy metabolism in pancreatic B-cell mitochondria. Studies with a non-metabolizable leucine analogue. Biochem J 219:189–196PubMed

26.

Heimberg H, De Vos A, Vandercammen A, Schaftingen E van, Pipeleers D, Schuit F (1993) Heterogeneity in glucose sensitivity among pancreatic beta-cells is correlated to differences in glucose phosphorylation rather than glucose transport. EMBO J 12:2873–2879PubMed

27.

Sener A, Malaisse-Lagae F, Malaisse WJ (1981) Stimulation of pancreatic islet metabolism and insulin release by a nonmetabolizable amino acid. Proc Natl Acad Sci USA 78:5460–5464PubMed

28.

Gao ZY, Li G, Najafi H, Wolf BA, Matschinsky FM (1999) Glucose regulation of glutaminolysis and its role in insulin secretion. Diabetes 48:1535–1542

29.

Li C, Najafi H, Daikhin Y et al. (2003) Regulation of leucine-stimulated insulin secretion and glutamine metabolism in isolated rat islets. J Biol Chem 278:2853–2858CrossRefPubMed

30.

Maechler P (2002) Mitochondria as the conductor of metabolic signals for insulin exocytosis in pancreatic beta-cells. Cell Mol Life Sci 59:1803–1818PubMed

31.

Broca C, Brennan L, Petit P, Newsholme P, Maechler P (2003) Mitochondria-derived glutamate at the interplay between branched-chain amino acid and glucose-induced insulin secretion. FEBS Lett 545:167–172CrossRefPubMed

32.

Fahien LA, MacDonald MJ, Kmiotek EH, Mertz RJ, Fahien CM (1988) Regulation of insulin release by factors that also modify glutamate dehydrogenase. J Biol Chem 263:13610–13614PubMed

33.

Malaisse-Lagae F, Sener A, Garcia-Morales P, Valverde I, Malaisse WJ (1982) The stimulus-secretion coupling of amino acid-induced insulin release. Influence of a nonmetabolized analog of leucine on the metabolism of glutamine in pancreatic islets. J Biol Chem 257:3754–3758PubMed

34.

Merglen A, Theander S, Rubi B, Chaffard G, Wollheim CB, Maechler P (2003) Glucose sensitivity and metabolism-secretion coupling studied during two-year continuous culture in INS-1E insulinoma cells. Endocrinology (in press)

35.

Asfari M, Janjic D, Meda P, Li G, Halban PA, Wollheim CB (1992) Establishment of 2-mercaptoethanol-dependent differentiated insulin-secreting cell lines. Endocrinology 130:167–178PubMed

36.

Pralong WF, Spat A, Wollheim CB (1994) Dynamic pacing of cell metabolism by intracellular Ca2+ transients. J Biol Chem 269:27310–27314PubMed

37.

Niwa H, Yamamura K, Miyazaki J (1991) Efficient selection for high-expression transfectants with a novel eukaryotic vector. Gene 108:193–199PubMed

38.

Miyake S, Makimura M, Kanegae Y (1996) Efficient generation of recombinant adenoviruses using adenovirus DNA-terminal protein complex and a cosmid bearing the full-length virus genome. Proc Natl Acad Sci USA 93:1320–1324PubMed

39.

Maechler P, Gjinovci A, Wollheim CB (2002) Implication of glutamate in the kinetics of insulin secretion in rat and mouse perfused pancreas. Diabetes 51 [Suppl 1]:S99–S102

40.

Maechler P, Jornot L, Wollheim CB (1999) Hydrogen peroxide alters mitochondrial activation and insulin secretion in pancreatic beta cells. J Biol Chem 274:27905–27913CrossRefPubMed

41.

Bustamante J, Lobo MV, Alonso FJ et al. (2001) An osmotic-sensitive taurine pool is localized in rat pancreatic islet cells containing glucagon and somatostatin. Am J Physiol Endocrinol Metab 281:E1275–E1285PubMed

42.

Maechler P, Kennedy ED, Pozzan T, Wollheim CB (1997) Mitochondrial activation directly triggers the exocytosis of insulin in permeabilized pancreatic beta-cells. EMBO J 16:3833–3841PubMed

43.

Maechler P, Wang H, Wollheim CB (1998) Continuous monitoring of ATP levels in living insulin secreting cells expressing cytosolic firefly luciferase. FEBS Lett 422:328–332PubMed

44.

Mosmann T (1983) Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxicity assays. J Immunol Methods 65:55–63PubMed

45.

Janjic D, Wollheim CB (1992) Islet cell metabolism is reflected by the MTT (tetrazolium) colorimetric assay. Diabetologia 35:482–485

46.

Schuit F, De Vos A, Farfari S et al. (1997) Metabolic fate of glucose in purified islet cells. Glucose-regulated anaplerosis in beta cells. J Biol Chem 272:18572–18579PubMed

47.

Lu D, Mulder H, Zhao P et al. (2002) 13C NMR isotopomer analysis reveals a connection between pyruvate cycling and glucose-stimulated insulin secretion (GSIS). Proc Natl Acad Sci USA 99:2708–2713CrossRefPubMed

48.

Owen OE, Kalhan SC, Hanson RW (2002) The key role of anaplerosis and cataplerosis for citric acid cycle function. J Biol Chem 277:30409–30412CrossRefPubMed

49.

Bai L, Zhang X, Ghishan FK (2003) Characterization of vesicular glutamate transporter in pancreatic alpha- and beta-cells and its regulation by glucose. Am J Physiol Gastrointest Liver Physiol 284:G808–G814PubMed

50.

Eto K, Yamashita T, Hirose K et al. (2003) Glucose metabolism and glutamate analog acutely alkalinize pH of insulin secretory vesicles of pancreatic beta-cells. Am J Physiol Endocrinol Metab 285:E262–E271PubMed

51.

Lenzen S, Schmidt W, Rustenbeck I, Panten U (1986) 2-ketoglutarate generation in pancreatic B-cell mitochondria regulates insulin secretory action of amino acids and 2-keto acids. Biosci Rep 6:163–169PubMed

52.

Kelly A, Li C, Gao Z, Stanley CA, Matschinsky FM (2002) Glutaminolysis and insulin secretion: from bedside to bench and back. Diabetes 51 [Suppl 3]:S421–S426

53.

Maechler P, Antinozzi PA, Wollheim CB (2000) Modulation of glutamate generation in mitochondria affects hormone secretion in INS-1E beta cells. IUBMB Life 50:27–31PubMed

54.

Sener A, Conget I, Rasschaert J et al. (1994) Insulinotropic action of glutamic acid dimethyl ester. Am J Physiol 267:E573–E584PubMed

55.

Michalik M, Nelson J, Erecinska M (1992) Glutamate production in islets of Langerhans: properties of phosphate-activated glutaminase. Metabolism 41:1319–1326PubMed

56.

Malaisse WJ, Sener A, Malaisse-Lagae F et al. (1982) The stimulus-secretion coupling of amino acid-induced insulin release. Metabolic response of pancreatic islets of L-glutamine and L-leucine. J Biol Chem 257:8731–8737PubMed

57.

Lenzen S, Rustenbeck I, Panten U (1984) Transamination of 3-phenylpyruvate in pancreatic B-cell mitochondria. J Biol Chem 259:2043–2046PubMed

58.

Lenzen S, Schmidt W, Panten U (1985) Transamination of neutral amino acids and 2-keto acids in pancreatic B-cell mitochondria. J Biol Chem 260:12629–12634PubMed

59.

Brennan L, Shine A, Hewage C et al. (2002) A nuclear magnetic resonance-based demonstration of substantial oxidative L-alanine metabolism and L-alanine-enhanced glucose metabolism in a clonal pancreatic beta-cell line: metabolism of L-alanine is important to the regulation of insulin secretion. Diabetes 51:1714–1721

60.

Brennan L, Corless M, Hewage C et al. (2003) 13C NMR analysis reveals a link between L-glutamine metabolism, D-glucose metabolism and gamma-glutamyl cycle activity in a clonal pancreatic beta-cell line. Diabetologia 46:1512–1521

61.

Tanizawa Y, Nakai K, Sasaki T et al. (2002) Unregulated elevation of glutamate dehydrogenase activity induces glutamine-stimulated insulin secretion: identification and characterization of a GLUD1 gene mutation and insulin secretion studies with MIN6 cells overexpressing the mutant glutamate dehydrogenase. Diabetes 51:712–717PubMed

62.

Boelens PG, Nijveldt RJ, Houdijk AP, Meijer S, Leeuwen PA van (2001) Glutamine alimentation in catabolic state. J Nutr 131 [Suppl]:2569S–2577S

63.

Henriksson J (1991) Effect of exercise on amino acid concentrations in skeletal muscle and plasma. J Exp Biol 160:149–165PubMed

64.

Ishihara H, Wang H, Drewes LR, Wollheim CB (1999) Overexpression of monocarboxylate transporter and lactate dehydrogenase alters insulin secretory responses to pyruvate and lactate in beta cells. J Clin Invest 104:1621–1629PubMed

65.

Otonkoski T, Kaminen N, Ustinov J, Lapatto R, Meissner T, Mayatepek E, Kere J, Sipila I (2003) Physical exercise-induced hyperinsulinemic hypoglycemia is an autosomal-dominant trait characterized by abnormal pyruvate-induced insulin release. Diabetes 52:199–204

Titel: Insulin secretion profiles are modified by overexpression of glutamate dehydrogenase in pancreatic islets
verfasst von: S. Carobbio
H. Ishihara
S. Fernandez-Pascual
C. Bartley
R. Martin-Del-Rio
P. Maechler
Publikationsdatum: 01.02.2004
Verlag: Springer-Verlag
Erschienen in: Diabetologia / Ausgabe 2/2004
Print ISSN: 0012-186X
Elektronische ISSN: 1432-0428
DOI: https://doi.org/10.1007/s00125-003-1306-2

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