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. 1983 May 15;212(2):385–392. doi: 10.1042/bj2120385

Insulin action on the glucose transport system in isolated cardiocytes from adult rat.

J Eckel, G Pandalis, H Reinauer
PMCID: PMC1152058  PMID: 6349617

Abstract

Calcium-tolerant myocytes from the adult rat heart were used to study the effects of insulin on the kinetics of myocardial 3-0-methylglucose transport at 37 degrees C. Insulin increased the initial velocity of sugar influx without affecting the equilibrium uptake values. Maximal stimulation averaged 50-80%, with a half-maximal response at an insulin concentration of 0.1 nM and maximal stimulation occurring at 1 nM. The onset of insulin action was preceded by a lag-phase of 20 s, reaching maximal action by 60 s. The Vmax. of the glucose transport system was increased from 160 to 287 nmol/min per 10(6) cells with an unaltered affinity. Neither extracellular nor intracellular calcium was found to be involved in the stimulatory action of insulin. Removal of intracellular magnesium resulted in a loss of insulin action. This study demonstrates that activation of the cardiac glucose transporter by insulin is due exclusively to an increase in the maximal velocity representing one of the very early effects of insulin on myocardial metabolism. The data suggest involvement of magnesium in the transmission of the insulin signal.

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Selected References

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

  1. Bihler I. Integration of glucose transport with metabolic and functional activity of heart muscle. Adv Myocardiol. 1980;2:3–15. [PubMed] [Google Scholar]
  2. Bowen-Pope D. F., Rubin H. Magnesium and calcium effects on uptake of hexoses and uridine by chick embryo fibroblasts. Proc Natl Acad Sci U S A. 1977 Apr;74(4):1585–1589. doi: 10.1073/pnas.74.4.1585. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. CROFFORD O. B., RENOLD A. E. GLUCOSE UPTAKE BY INCUBATED RAT EPIDIDYMAL ADIPOSE TISSUE. RATE-LIMITING STEPS AND SITE OF INSULIN ACTION. J Biol Chem. 1965 Jan;240:14–21. [PubMed] [Google Scholar]
  4. Cheung J. Y., Conover C., Regen D. M., Whitfield C. F., Morgan H. E. Effect of insulin on kinetics of sugar transport in heart muscle. Am J Physiol. 1978 Jan;234(1):E70–E78. doi: 10.1152/ajpendo.1978.234.1.E70. [DOI] [PubMed] [Google Scholar]
  5. Ciaraldi T. P., Olefsky J. M. Coupling of insulin receptors to glucose transport: a temperature-dependent time lag in activation of glucose transport. Arch Biochem Biophys. 1979 Mar;193(1):221–231. doi: 10.1016/0003-9861(79)90026-2. [DOI] [PubMed] [Google Scholar]
  6. Cuatrecasas P., Hollenberg M. D., Chang K. J., Bennett V. Hormone receptor complexes and their modulation of membrane function. Recent Prog Horm Res. 1975;31:37–94. doi: 10.1016/b978-0-12-571131-9.50006-2. [DOI] [PubMed] [Google Scholar]
  7. Cushman S. W., Wardzala L. J. Potential mechanism of insulin action on glucose transport in the isolated rat adipose cell. Apparent translocation of intracellular transport systems to the plasma membrane. J Biol Chem. 1980 May 25;255(10):4758–4762. [PubMed] [Google Scholar]
  8. Czech M. P. Insulin action and the regulation of hexose transport. Diabetes. 1980 May;29(5):399–409. doi: 10.2337/diab.29.5.399. [DOI] [PubMed] [Google Scholar]
  9. Czech M. P. Molecular basis of insulin action. Annu Rev Biochem. 1977;46:359–384. doi: 10.1146/annurev.bi.46.070177.002043. [DOI] [PubMed] [Google Scholar]
  10. Dow J. W., Harding N. G., Powell T. Isolated cardiac myocytes. I. Preparation of adult myocytes and their homology with the intact tissue. Cardiovasc Res. 1981 Sep;15(9):483–514. doi: 10.1093/cvr/15.9.483. [DOI] [PubMed] [Google Scholar]
  11. Eckel J., Offermann A., Reinauer H. Insulin receptors on isolated heart cells: effect of temperature and hydrolytic enzymes. Basic Res Cardiol. 1982 May-Jun;77(3):323–332. doi: 10.1007/BF01908047. [DOI] [PubMed] [Google Scholar]
  12. Eckel J., Reinauer H. Characteristics of insulin receptors in the heart muscle: binding of insulin to isolated muscle cells from adult rat heart. Biochim Biophys Acta. 1980 May 22;629(3):510–521. doi: 10.1016/0304-4165(80)90156-7. [DOI] [PubMed] [Google Scholar]
  13. Eckel J., Reinauer H. Effect of vinblastine on the insulin-receptor interaction in mammalian heart muscle. Biochem Biophys Res Commun. 1980 Feb 27;92(4):1403–1408. doi: 10.1016/0006-291x(80)90442-8. [DOI] [PubMed] [Google Scholar]
  14. Eckel J., Reinauer H. The fate of insulin in cardiac muscle. Studies on isolated muscle cells from adult rat heart. Biochem J. 1982 Sep 15;206(3):655–662. doi: 10.1042/bj2060655. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Farmer B. B., Harris R. A., Jolly W. W., Hathaway D. R., Katzberg A., Watanabe A. M., Whitlow A. L., Besch H. R., Jr Isolation and characterization of adult rat hearts cells. Arch Biochem Biophys. 1977 Mar;179(2):545–558. doi: 10.1016/0003-9861(77)90143-6. [DOI] [PubMed] [Google Scholar]
  16. Fischer U. Die Insulinwirkung auf die transmembrane Glukosebilanz des iolierten perfundierten Rattenherzen. II. Die Wirkung von Insulin auf die kinetischen Parameter der Glukoseaufnahme. Acta Biol Med Ger. 1971;26(1):87–99. [PubMed] [Google Scholar]
  17. Gerards P., Graf W., Kammermeier Glucose transfer studies in isolated cardiocytes of adult rats. J Mol Cell Cardiol. 1982 Mar;14(3):141–149. doi: 10.1016/0022-2828(82)90112-2. [DOI] [PubMed] [Google Scholar]
  18. Gould M. K., Chaudry I. H. The action of insulin on glucose uptake by isolated rat soleus muscle. I. Effects of cations. Biochim Biophys Acta. 1970 Aug 14;215(2):249–257. doi: 10.1016/0304-4165(70)90022-x. [DOI] [PubMed] [Google Scholar]
  19. Hainaut K., Desmedt J. E. Calcium ionophore A23187 potentiates twitch and intracellular calcium release in single muscle fibres. Nature. 1974 Nov 29;252(5482):407–408. doi: 10.1038/252407a0. [DOI] [PubMed] [Google Scholar]
  20. Haworth R. A., Hunter D. R., Berkoff H. A. Insulin stimulates deoxyglucose transport in adult rat heart cells in the absence of Ca2+. FEBS Lett. 1982 May 3;141(1):37–40. doi: 10.1016/0014-5793(82)80010-0. [DOI] [PubMed] [Google Scholar]
  21. Haworth R. A., Hunter D. R., Berkoff H. A. The isolation of Ca2+-resistant myocytes from the adult rat. J Mol Cell Cardiol. 1980 Jul;12(7):715–723. doi: 10.1016/0022-2828(80)90101-7. [DOI] [PubMed] [Google Scholar]
  22. Kao R. L., Christman E. W., Luh S. L., Krauhs J. M., Tyers G. F., Williams E. H. The effects of insulin and anoxia on the metabolism of isolated mature rat cardiac myocytes. Arch Biochem Biophys. 1980 Sep;203(2):587–599. doi: 10.1016/0003-9861(80)90216-7. [DOI] [PubMed] [Google Scholar]
  23. Karnieli E., Zarnowski M. J., Hissin P. J., Simpson I. A., Salans L. B., Cushman S. W. Insulin-stimulated translocation of glucose transport systems in the isolated rat adipose cell. Time course, reversal, insulin concentration dependency, and relationship to glucose transport activity. J Biol Chem. 1981 May 25;256(10):4772–4777. [PubMed] [Google Scholar]
  24. LEVINE R., GOLDSTEIN M. S., HUDDLESTUN B., KLEIN S. P. Action of insulin on the 'permeability' of cells to free hexoses, as studied by its effect on the distribution of galactose. Am J Physiol. 1950 Oct;163(1):70–76. doi: 10.1152/ajplegacy.1950.163.1.70. [DOI] [PubMed] [Google Scholar]
  25. Lundin A., Richardsson A., Thore A. Continous monitoring of ATP-converting reactions by purified firefly luciferase. Anal Biochem. 1976 Oct;75(2):611–620. doi: 10.1016/0003-2697(76)90116-0. [DOI] [PubMed] [Google Scholar]
  26. MORGAN H. E., HENDERSON M. J., REGEN D. M., PARK C. R. Regulation of glucose uptake in muscle. I. The effects of insulin and anoxia on glucose transport and phosphorylation in the isolated, perfused heart of normal rats. J Biol Chem. 1961 Feb;236:253–261. [PubMed] [Google Scholar]
  27. MORGAN H. E., RANDLE P. J., REGEN D. M. Regulation of glucose uptake by muscle. 3. The effects of insulin, anoxia, salicylate and 2:4-dinitrophenol on membrane transport and intracellular phosphorylation of glucose in the isolated rat heart. Biochem J. 1959 Dec;73:573–579. doi: 10.1042/bj0730573. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. MORGAN H. E., REGEN D. M., PARK C. R. IDENTIFICATION OF A MOBILE CARRIER-MEDIATED SUGAR TRANSPORT SYSTEM IN MUSCLE. J Biol Chem. 1964 Feb;239:369–374. [PubMed] [Google Scholar]
  29. Montini J., Bagby G. J., Burns A. H., Spitzer J. J. Exogenous substrate utilization in Ca2+-tolerant myocytes from adult rat hearts. Am J Physiol. 1981 Apr;240(4):H659–H663. doi: 10.1152/ajpheart.1981.240.4.H659. [DOI] [PubMed] [Google Scholar]
  30. Murray J. J., Reed P. W., Fay F. S. Contraction of isolated smooth muscle cells by inophore A23187. Proc Natl Acad Sci U S A. 1975 Nov;72(11):4459–4463. doi: 10.1073/pnas.72.11.4459. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. PARK C. R., REINWEIN D., HENDERSON M. J., CADENAS E., MORGAN H. E. The action of insulin on the transport of glucose through the cell membrane. Am J Med. 1959 May;26(5):674–684. doi: 10.1016/0002-9343(59)90227-x. [DOI] [PubMed] [Google Scholar]
  32. Reeves J. P. Ca2+ ions and the stimulation of 3-O-methylglucose transport by uncouplers in rat thymocytes. J Biol Chem. 1977 Jul 25;252(14):4876–4881. [PubMed] [Google Scholar]
  33. Schudt C., Gaertner U., Pette D. Insulin action on glucose transport and calcium fluxes in developing muscle cells in vitro. Eur J Biochem. 1976 Sep;68(1):103–111. doi: 10.1111/j.1432-1033.1976.tb10768.x. [DOI] [PubMed] [Google Scholar]
  34. Shine K. I., Douglas A. M. Magnesium effects in rabbit ventricle. Am J Physiol. 1975 May;228(5):1545–1554. doi: 10.1152/ajplegacy.1975.228.5.1545. [DOI] [PubMed] [Google Scholar]
  35. Shine K. I. Myocardial effects of magnesium. Am J Physiol. 1979 Oct;237(4):H413–H423. doi: 10.1152/ajpheart.1979.237.4.H413. [DOI] [PubMed] [Google Scholar]
  36. Siegel J., Olefsky J. M. Role of intracellular energy in insulin's ability to activate 3-O-methylglucose transport by rat adipocytes. Biochemistry. 1980 May 13;19(10):2183–2190. doi: 10.1021/bi00551a029. [DOI] [PubMed] [Google Scholar]
  37. Suzuki K., Kono T. Evidence that insulin causes translocation of glucose transport activity to the plasma membrane from an intracellular storage site. Proc Natl Acad Sci U S A. 1980 May;77(5):2542–2545. doi: 10.1073/pnas.77.5.2542. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Whitesell R. R., Gliemann J. Kinetic parameters of transport of 3-O-methylglucose and glucose in adipocytes. J Biol Chem. 1979 Jun 25;254(12):5276–5283. [PubMed] [Google Scholar]
  39. Yu K. T., Gould M. K. Stimulation of sugar transport in rat soleus muscle by prolonged cooling at 0 degrees C. Diabetologia. 1981 Nov;21(5):482–488. doi: 10.1007/BF00257790. [DOI] [PubMed] [Google Scholar]

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