Abstract
The effects of the muscarinic agonist oxotremorine-m (oxo-m) on insulin secretion, K+-permeability and electrical activity from isolated mouse pancreatic islets were studied. Oxo-m potentiated glucose-induced insulin secretion in a dose-dependent manner, saturating at ca. 10 μm. At 11.2 mm glucose, oxo-m (0.1 and 10 μm) had two distinct effects on β-cell electrical activity. Both concentrations increased the steadystate burst frequency, however, at 10 μm an initial and transient polarization was measured, and the subsequent activity was accompanied by a slight depolarization. The polarizing effect of oxo-m was almost completely suppressed by charybdotoxin (ChTX), a blocker of the large conductance (maxi) [Ca2+] i -activated potassium channel (K(Ca)). In the presence of 11.2 mm glucose, oxo-m (50 μm) provoked a significant and transient increase in the 86Rb efflux from perifused islets. This effect was inhibited by ChTX. ChTX also potentiated oxo-m stimulated insulin secretion in the presence of glucose. Finally, the balance between the polarizing and depolarizing effects of oxo-m was variable in different islets and depended on glucose concentration. Insulin secretion stimulated by oxo-m in the presence of glucose was more closely correlated to the agonist induced increase in burst frequency than to an increase in plateau fraction. We conclude that muscarinic stimulation has at least two effects on β-cell electrical activity, an initial hyperpolarization, owing to activation of K(Ca) channels, followed by depolarization and high-frequency bursts, proposed to reflect the activation of a current sensitive to the depletion of intracellular Ca2+ stores (CRAC).
Similar content being viewed by others
References
Ämmälä, C., Larsson, O., Berggren, P-O., Bokvist, K., Juntti-Berggren, L., Kindmark, H., Rorsman, P. 1991. Inositol triphosphate-dependent periodic activation of a Ca2+-activated K+ conductance in glucose-stimulated pancreatic B-cells. Nature 353:849–852
Atwater, I., Carroll, P.C., Li, M.X. 1989. Electrophysiology of the pancreatic B-cell. In: Molecular and Cellular Biology of Diabetes Mellitus, B. Draznin, S. Melmed and D. LeRoith, editors. pp. 49–68. Alan R. Liss, New York
Atwater, I., Dawson, C., Scott, A., Eddlestone, G.T., Rojas, E. 1980. The nature of the oscillatory behavior in electrical activity from pancreatic β-cell. Horm. Metab. Res., Suppl. 10, 100–107
Atwater, I., Ribalet, B., Rojas, E. 1978. Cyclic changes in potential and resistance of the B-cell membrane induced by glucose in islets of Langerhans from mouse. J. Physiol. 278:117–139
Bertram, R., Smolen, P., Sherman, A., Mears, D., Atwater, I., Martin, F., Soria, B. 1995. A role for calcium release activated current (CRAC) in cholinergic modulation of electrical activity in pancreatic B-cells. Biophys. J. (in press)
Birdsall, N.J.M., Burgen, A.S.V., Hulme, E.C. 1978. The binding of agonists to brain muscarinic receptors. Mol. Pharmacol. 14:723–736
Boschero, A.C., Malaisse, W.J. 1979. Stimulus-secretion coupling of glucose-induced insulin release. XXIX. Regulation of 86Rb efflux from perifused islets. Am. J. Physiol. 236:E139-E146
Boschero, A.C., Szpak-Glasman, M., Carneiro, E.M., Bordin, S., Paul, I., Rojas, E., Atwater, I. 1995. Potentiation of glucose-induced insulin release from rat pancreatic islets by oxotremorine-m involves M3 muscarinic cholinergic receptors. Am. J. Physiol. 268:E336-E342
Clementi, E., Scheer, H., Zacchetti, D., Fasolato, C., Pozzan, T., Meldolesi, J. 1992. Receptor-activated Ca2+ influx. J. Biol. Chem. 267:2164–2172
Cook, D.L., Crill, W.E., Porte, D., Jr. 1981. Glucose and acetylcholine have different effects on the plateau pacemaker of pancreatic islet cells. Diabetes 30:558–561
Dunlop, M., Shaw, M., Dimitriadis, E., Gurtler, V., Wark, J., Larkins, R.G. 1988. Evidence that muscarinic receptors in islets cells are not coupled functionally to adenylate cyclase through the inhibitory guanine nucleotide binding protein (Ni). Horm. Metabol. Res. 20:150–153
Gagerman, E., Idahl, L.A., Meissner, H.P., Täljedal, I-B. 1978. Insulin release, cGMP, cAMP, and membrane potential in acetylcholine stimulated islets. Am. J. Physiol. 235:E493-E500
Henquin, J.C., Garcia, M-C., Bozem, M., Hermans, M.P., Nenquim, M. 1988. Muscarinic control of pancreatic B cell function involves sodium-dependent depolarization and calcium influx. Endocrinology 122:21134–2142
Henquin, J.C., Nenquin, M. 1988. The muscarinic receptor subtype in mouse pancreatic B-cells. FEBS Lett. 236:89–92
Hermans, M.P., Schmeer, W., Henquin, J.C. 1987. Modulation of the effect of acetylcholine on insulin release by the membrane potential of B cells. Endocrinology 120:1765–1773
Hiriart, M., Ramirez-Medeles. 1993. Muscarinic modulation of insulin secretion by single pancreatic beta-cells. Mol Cell. Endocrinol. 93:63–69
Hoth, M., Penner, R. 1992. Depletion of intracellular calcium stores activates a calcium current in mast cells. Nature 355:353–356
Kukuljan, M., Goncalves, A.A., Atwater, I. 1991. Charybdotoxin sensitive K(Ca) channel is not involved in glucose-induced electrical activity in pancreatic B-cells. J. Membrane Biol. 119:187–195
Lazrak, A., Peracchia, C. 1993. Gap junction gating sensitivity to physiological internal calcium regardless of pH in Novikoff hepatoma cells. Biophys. J. 65:2002–2012
Leech, C.A., Holz, IV, G.G., Habener, J.F. 1994. Voltage-independent calcium channels mediate slow oscillations of cytosolic calcium that are glucose dependent in pancreatic B-cells. Endocrinology 135:365–372
Loewenstein, W.R., Rose, B. 1978. Calcium in (junctional) intercellular communication and a thought on its behavior in intracellular communication. Ann. N.Y. Acad. Sci. 307:285–305
Malaisse, W.J. 1986. Stimulus-secretion coupling in the pancreatic B-cell. The cholinergic pathway for insulin release. Diabetes Metabolism Reviews. Vol 3. pp 243–259. Wiley & Sons, New York
Parekh, A.B., Terlau, H., Stuhmer, W. 1993. Depletion of InsP3 stores activates a Ca2+ and K+ current by means of a phosphatase and a diffusible messenger. Nature 364:814–818
Putney, J.W. 1990. Capacitative calcium entry revisited. Cell Calcium 11:611–624
Randriamampita, C., Tsien, R.Y. 1993. Emptying of intracellular Ca2+ stores releases a novel small messenger that stimulates Ca2+ influx. Nature 364:809–814
Rojas, E., Carroll, B.P., Ricordi, C., Boschero, A.C., Stojilkovic, S.S., Atwater, I. 1994. Control of cytosolic free-calcium in cultured human pancreatic β-cells occurs by external calcium-dependent and independent mechanisms. Endocrinology 134:1771–1781
Rojas, E., Hidalgo, J., Carroll, P.B., Li, M.X., Atwater, I. 1990. A new class of calcium channels activated by glucose in human pancreatic β-cells. FEBS Lett. 261:265–270
Rojas, E., Santos, R.M., Atwater, I. 1990. Role of membrane receptors in stimulus-secretion coupling. In: Transduction in Biological Systems. C. Hidalgo, J. Bacigalupo, E. Jaimovich, J. Vergara, editors, pp. 101–122. Plenum, New York
Sanchez-Andres, J.V., Ripoll, C., Soria, B. 1988. Evidence that muscarinic potentiation of insulin release is initiated by an early transient calcium entry. FEBS Lett. 231:143–147
Santos, R.M., Rojas, E. 1989. Muscarinic receptor modulation of glucose-induced electrical activity in mouse pancreatic B-cells. FEBS Lett. 249:411–417
Satin, L.S., Tavalin, S.J., Kinard, T.A., Teague, J. 1995. Contribution of Land non-L-type calcium channels to voltage-gated calcium current and glucose-dependent insulin secretion in H1T-T15 cells. Endocrinology 136:4589–4601
Scott, A.M., Atwater, I., Rojas, E. 1981. A method for the simultaneous measurement of insulin release and B-cell membrane potential in single mouse islets of Langerhans. Diabetologia 21:470–475
Sethy, V.H., Francis, J.W. 1990. Pharmacokinetics of muscarinic cholinergic drags as determined by ex vivo (3H)-oxotremorine-M binding. J. Pharmacol. Methods 23:285–296
Sherman, A. 1995. Contributions of Modeling to Understanding Stimulus-Secretion Coupling in Pancreatic B-cells. Am. J. Physiol. (in press)
Tabcharani, J.A., Misler, S. 1989. Ca2+-activated K+ channel in rat pancreatic B cells: permeation, gatin blockade by cations. Biochem. Biophys. Acta 982:62–72
Takemura, H., Hughes, A.R., Thastrup, O., Putney, J.J.W. 1989. Activation of calcium entry by the tumor promotor thapsigargin in parotid acinar cells. J. Biol. Chem. 264:12266–12271
Verspohl, E.J., Tacke, R., Mutschier, E., Lambrecht, G. 1990. Muscarinic receptor subtypes in rat pancreatic islets: binding and functional studies. eur. J. Pharmacol. 178:303–311
Worley III, J.F., McIntyre, B., Spencer, B., Mertz, R.J., Roe, M.W., Dukes, I.D. 1994. Endoplasmic reticulum calcium store regulates membrane potential in mouse islet B-cells. J. Biol. Chem. 269:14359–14362
Author information
Authors and Affiliations
Additional information
We thank Dr. Mirta Szpak-Glasman for collaboration during the data analysis and Drs. David Mears, Arthur Sherman, Paul Smolen and Richard Bertram for critical reading of the manuscript. This work was supported in part by Conselho Nacional de Desenvolvimento Cientifico e Tecnologico (CNPq), Coordenadoria de Aperfeicoamento do Ensino Superior (CAPES), and Fundacao de Amparo a Pesquisa do Estado de Sao Paulo (FAPESP), Brazil.
Rights and permissions
About this article
Cite this article
Bordin, S., Boschero, A.C., Carneiro, E.M. et al. Ionic mechanisms involved in the regulation of insulin secretion by muscarinic agonists. J. Membarin Biol. 148, 177–184 (1995). https://doi.org/10.1007/BF00207273
Received:
Revised:
Issue Date:
DOI: https://doi.org/10.1007/BF00207273