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. 1981 Aug;68(2):517–524. doi: 10.1172/JCI110283

Fluctuations of calcium, phosphorus, sodium, potassium, and chlorine in single alpha and beta cells during glucose perifusion of rat islets.

R K Kalkhoff, K A Siegesmund
PMCID: PMC370826  PMID: 7021594

Abstract

To study the relationship between islet hormonal secretion and intracellular content of five elements, a rat islet perifusion technique was used in 24 paired experiments. Control and experimental chambers each containing 100 islets, received 2.8 and 16.7 mM D-glucose, respectively. Effluent was collected frequently for hormone measurements. At eight different time intervals form 0--30 min islets were fixed and prepared for scanning electron microscopy. Over 900 unobscured alpha and beta cells were selected by size and shape criteria. Energy dispersive x-ray analysis was applied to each single cell to determine relative content of calcium (Ca), potassium (K), sodium (Na), chlorine (Cl), and phosphorus (P). Experimental chambers exhibited typical acute (0--9 min) and second phase (10--30 min) insulin secretion in association with suppression of glucagon release after 10 min. At 2 min an abrupt upward K spike in both alpha and beta cells was followed at 3--4 min with a 1.5- to 2-fold rise of Ca and a reciprocal decrease in K, Na, Cl, and P. From 3 to 30 min biphasic insulin secretion. Reduced alpha cell calcium after 6 min preceded suppression of glucagon secretion. After 2 min K related inversely to Ca content in both alpha and beta cells. These results could not be reproduced when D-galactose was substituted for D-glucose. We conclude that sequential changes of Ca content that are reciprocally related to K are predictive of beta cell insulin release and suppression of alpha cell glucagon secretion.

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  1. Ashby J. P., Speake R. N. Insulin and glucagon secretion from isolated islets of Langerhans. The effects of calcium ionophores. Biochem J. 1975 Jul;150(1):89–96. [PMC free article] [PubMed] [Google Scholar]
  2. Berglund O., Sehlin J. Defective regulation of Cl- permeability in islets of diabetic mice [C57BL/KsJ(db/db)]. Diabetes. 1980 Feb;29(2):151–155. doi: 10.2337/diab.29.2.151. [DOI] [PubMed] [Google Scholar]
  3. Boschero A. C., Kawazu S., Duncan G., Malaisse W. J. Effect of glucose on K+ handling by pancreatic islets. FEBS Lett. 1977 Nov 1;83(1):151–154. doi: 10.1016/0014-5793(77)80662-5. [DOI] [PubMed] [Google Scholar]
  4. Boschero A. C., Malaisse W. J. Effect of glucose on 22Na+ efflux in pancreatic islets. Arch Int Physiol Biochim. 1978 Aug;86(3):479–485. doi: 10.3109/13813457809055918. [DOI] [PubMed] [Google Scholar]
  5. Bukowiecki L., Freinkel N. Relationship between efflux of ionic calcium and phosphorus during excitation of pancreatic islets with glucose. Biochim Biophys Acta. 1976 Jun 4;436(1):190–198. doi: 10.1016/0005-2736(76)90230-3. [DOI] [PubMed] [Google Scholar]
  6. Charles M. A., Lawecki J., Pictet R., Grodsky G. M. Insulin secretion. Interrelationships of glucose, cyclic adenosine 3:5-monophosphate, and calcium. J Biol Chem. 1975 Aug 10;250(15):6134–6140. [PubMed] [Google Scholar]
  7. Dean P. M., Matthews E. K. Electrical activity in pancreatic islet cells: effect of ions. J Physiol. 1970 Sep;210(2):265–275. doi: 10.1113/jphysiol.1970.sp009208. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Devis G., Somers G., Malaisse W. J. Stimulation of insulin release by calcium. Biochem Biophys Res Commun. 1975 Nov 17;67(2):525–529. doi: 10.1016/0006-291x(75)90843-8. [DOI] [PubMed] [Google Scholar]
  9. Formby B., Capito K., Egeberg J., Hedeskov C. J. Ca-activated ATPase activity in subcellular fractions of mouse pancreatic islets. Am J Physiol. 1976 Feb;230(2):441–448. doi: 10.1152/ajplegacy.1976.230.2.441. [DOI] [PubMed] [Google Scholar]
  10. Frankel B. J., Imagawa W. T., O'Connor M. D., Lundquist I., Kromhout J. A., Fanska R. E., Grodsky G. M. Glucose-stimulated 45Calcium efflux from isolated rat pancreatic islets. J Clin Invest. 1978 Sep;62(3):525–531. doi: 10.1172/JCI109156. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Freinkel N., Pedley K. C., Wooding P., Dawson R. M. Localization of inorganic phosphate in the pancreatic B cell and its loss on glucose stimulation. Science. 1978 Sep 22;201(4361):1124–1126. doi: 10.1126/science.356269. [DOI] [PubMed] [Google Scholar]
  12. Freinkel N., Younsi C. E., Bonnar J., Dawson R. M. Rapid transient efflux of phosphate ions from pancreatic islets as an early action of insulin secretagogues. J Clin Invest. 1974 Nov;54(5):1179–1189. doi: 10.1172/JCI107861. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Grodsky G. M., Bennett L. L. Cation requirements for insulin secretion in the isolated perfused pancreas. Diabetes. 1966 Dec;15(12):910–913. doi: 10.2337/diab.15.12.910. [DOI] [PubMed] [Google Scholar]
  14. Hellman B. Stimulation of insulin release after raising extracellular calcium. FEBS Lett. 1976 Mar 15;63(1):125–128. doi: 10.1016/0014-5793(76)80208-6. [DOI] [PubMed] [Google Scholar]
  15. Henquin J. C. D-glucose inhibits potassium efflux from pancreatic islet cells. Nature. 1978 Jan 19;271(5642):271–273. doi: 10.1038/271271a0. [DOI] [PubMed] [Google Scholar]
  16. Henquin J. C. Opposite effects of intracellular Ca2+ and glucose on K+ permeability of pancreatic islet cells. Nature. 1979 Jul 5;280(5717):66–68. doi: 10.1038/280066a0. [DOI] [PubMed] [Google Scholar]
  17. Kalkhoff R. K., Kim H. J. Effects of pregnancy on insulin and glucagon secretion by perifused rat pancreatic islets. Endocrinology. 1978 Feb;102(2):623–631. doi: 10.1210/endo-102-2-623. [DOI] [PubMed] [Google Scholar]
  18. Kawazu S., Boschero A. C., Delcroix C., Malaisse W. J. The stimulus-secretion coupling of glucose-induced insulin release. XXVIII. Effect of glucose on Na+ fluxes in isolated islets. Pflugers Arch. 1978 Jul 18;375(2):197–206. doi: 10.1007/BF00584244. [DOI] [PubMed] [Google Scholar]
  19. Kikuchi M., Wollheim C. B., Cuendet G. S., Renold A. E., Sharp G. W. Studies on the dual effects of glucose on 45Ca++ efflux from isolated rat islets. Endocrinology. 1978 May;102(5):1339–1349. doi: 10.1210/endo-102-5-1339. [DOI] [PubMed] [Google Scholar]
  20. Lacy P. E. Endocrine secretory mechanisms. A review. Am J Pathol. 1975 Apr;79(1):170–188. [PMC free article] [PubMed] [Google Scholar]
  21. Lacy P. E., Kostianovsky M. Method for the isolation of intact islets of Langerhans from the rat pancreas. Diabetes. 1967 Jan;16(1):35–39. doi: 10.2337/diab.16.1.35. [DOI] [PubMed] [Google Scholar]
  22. Lacy P. E., Walker M. M., Fink C. J. Perifusion of isolated rat islets in vitro. Participation of the microtubular system in the biphasic release of insulin. Diabetes. 1972 Oct;21(10):987–998. doi: 10.2337/diab.21.10.987. [DOI] [PubMed] [Google Scholar]
  23. Lechene C. Electron probe microanalysis of biological soft tissues: principle and technique. Fed Proc. 1980 Sep;39(11):2871–2880. [PubMed] [Google Scholar]
  24. Leclercq-Meyer V., Marchand J., Malaisse W. J. The effect of calcium and magnesium on glucagon secretion. Endocrinology. 1973 Dec;93(6):1360–1370. doi: 10.1210/endo-93-6-1360. [DOI] [PubMed] [Google Scholar]
  25. Levin S. R., Kasson B. G., Driessen J. F. Adenosine triphosphatases of rat pancreatic islets: comparison with those of rat kidney. J Clin Invest. 1978 Sep;62(3):692–701. doi: 10.1172/JCI109177. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Malaisse-Lagae F., Malaisse W. J. The stimulus-secretion coupling of glucose-induced insulin release. 3. Uptake of 45 calcium by isolated islets of Langerhans. Endocrinology. 1971 Jan;88(1):72–80. doi: 10.1210/endo-88-1-72. [DOI] [PubMed] [Google Scholar]
  27. Malaisse W. J., Brisson G. R., Baird L. E. Stimulus-secretion coupling of glucose-induced insulin release. X. Effect of glucose on 45 Ca efflux from perifused islets. Am J Physiol. 1973 Feb;224(2):389–394. doi: 10.1152/ajplegacy.1973.224.2.389. [DOI] [PubMed] [Google Scholar]
  28. Martonosi A. N. Calcium pumps: introduction. Fed Proc. 1980 May 15;39(7):2401–2402. [PubMed] [Google Scholar]
  29. Meissner H. P., Preissler M. Glucose-induced changes of the membrane potential of pancreatic B-cells: their significance for the regulation of insulin release. Adv Exp Med Biol. 1979;119:97–107. doi: 10.1007/978-1-4615-9110-8_15. [DOI] [PubMed] [Google Scholar]
  30. Meissner H. P., Schmelz H. Membrane potential of beta-cells in pancreatic islets. Pflugers Arch. 1974;351(3):195–206. doi: 10.1007/BF00586918. [DOI] [PubMed] [Google Scholar]
  31. Naber S. P., McDaniel M. L., Lacy P. E. The effect of glucose on the acute uptake and efflux of calcium-45 in isolated rat islets. Endocrinology. 1977 Sep;101(3):686–693. doi: 10.1210/endo-101-3-686. [DOI] [PubMed] [Google Scholar]
  32. Orci L., Malaisse W. Hypothesis: single and chain release of insulin secretory granules is related to anionic transport at exocytotic sites. Diabetes. 1980 Nov;29(11):943–944. doi: 10.2337/diab.29.11.943. [DOI] [PubMed] [Google Scholar]
  33. Pace C. S. Activation of Na channels in islet cells: metabolic and secretory effects. Am J Physiol. 1979 Aug;237(2):E130–E135. doi: 10.1152/ajpendo.1979.237.2.E130. [DOI] [PubMed] [Google Scholar]
  34. Pierce M., Freinkel N., Dawson R. M., Asplund K., Bukowiecki L. 32P-labeling patterns in rat pancreatic islets: tissue source of the radiophosphate released after glucose stimulation. Endocrinology. 1978 Sep;103(3):971–977. doi: 10.1210/endo-103-3-971. [DOI] [PubMed] [Google Scholar]
  35. Ribalet B., Beigelman P. M. Cyclic variation of K+ conductance in pancreatic beta-cells: Ca2+ and voltage dependence. Am J Physiol. 1979 Sep;237(3):C137–C146. doi: 10.1152/ajpcell.1979.237.3.C137. [DOI] [PubMed] [Google Scholar]
  36. Rocha D. M., Faloona G. R., Unger R. H. Glucagon-stimulating activity of 20 amino acids in dogs. J Clin Invest. 1972 Sep;51(9):2346–2351. doi: 10.1172/JCI107046. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Sehlin J., Taljedal I. B. Glucose-induced decrease in Rb+ permeability in pancreatic beta cells. Nature. 1975 Feb 20;253(5493):635–636. doi: 10.1038/253635a0. [DOI] [PubMed] [Google Scholar]
  38. Sehlin J., Täljedal I. B. Sodium uptake by microdissected pancreatic islets: effects of ouabain and chloromercuribenzene-p-sulphonic acid. FEBS Lett. 1974 Feb 15;39(2):209–213. doi: 10.1016/0014-5793(74)80052-9. [DOI] [PubMed] [Google Scholar]
  39. Siegel E. G., Wollheim C. B., Renold A. E., Sharp G. W. Evidence for the involvement of Na/Ca exchange in glucose-induced insulin release from rat pancreatic islets. J Clin Invest. 1980 Nov;66(5):996–1003. doi: 10.1172/JCI109969. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Van Obberghen E., Somers G., Devis G., Ravazzola M., Malaisse-Lagae F., Orci L., Malaisse W. J. Dynamics of insulin release and microtubular-microfilamentous system. VII. Do microfilaments provide the motive force for the translocation and extrusion of beta granules? Diabetes. 1975 Oct;24(10):892–901. doi: 10.2337/diab.24.10.892. [DOI] [PubMed] [Google Scholar]
  41. Wollheim C. B., Kikuchi M., Renold A. E., Sharp G. W. The roles of intracellular and extracellular Ca++ in glucose-stimulated biphasic insulin release by rat islets. J Clin Invest. 1978 Aug;62(2):451–458. doi: 10.1172/JCI109146. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. Zimny M. L., Blackard W. G. The surface structure of isolated pancreatic islet cells. Cell Tissue Res. 1975 Dec 18;164(4):467–471. doi: 10.1007/BF00219938. [DOI] [PubMed] [Google Scholar]

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