Skip to main content

Regulation by Insulin and Insulin-Like Growth Factor of 2-Deoxyglucose Uptake in Primary Ependymal Cell Cultures

Neurochemical Research volume 29, pages 127–134 (2004)Cite this article

Abstract

Ependymal cells have been reported to express the facilitative glucose carriers GLUT1, GLUT2, and GLUT4, as well as glucokinase. They are therefore speculated to be part of the cerebral glucose sensing system and may also respond to insulin with alterations in their glucose uptake rate. A cell culture model was employed to study the functional status of ependymal insulin-regulated glucose uptake in vitro. Insulin increased the uptake of the model substrate 2-deoxyglucose (2-DG) dependent on the insulin concentration. This was due to a near doubling of the maximal 2-DG uptake rate. Insulin-like growth factor (IGF-1) was at least 10 times more potent than insulin in stimulating the rate of ependymal 2-DG uptake, suggesting that IGF-1, rather than insulin, is the physiological agonist regulating glucose transport in ependymal cells. The predominant glucose transporter in ependymal cell cultures was found to be GLUT1, which is apparently regulated by IGF-1 in ependymal cells.

This is a preview of subscription content, access via your institution.

Access options

Buy single article

Instant access to the full article PDF.

43,34 €

Price includes VAT (Finland)
Tax calculation will be finalised during checkout.

REFERENCES

  1. Joost, H. G. and Thorens, B. 2001. The extended GLUT-family of sugar/polyol transport facilitators: Nomenclature, sequence characteristics, and potential function of its novel members. Mol. Membr. Biol. 18:247–256.

    Google Scholar 

  2. Dwyer, D. S., Vannucci, S. J., and Simpson, I. A. 2002. Expression, regulation, and functional role of glucose transporters (GLUTs) in brain. Int. Rev. Neurobiol. 51:159–188.

    Google Scholar 

  3. Birnbaum, M. J., Haspel, H. C., and Rosen, O. M. 1986. Cloning and characterization of a cDNA encoding the rat brain glucose transporter protein. Proc. Natl. Acad. Sci. USA 83:5784–5788.

    Google Scholar 

  4. Klepper, J. and Voit, T. 2002. Facilitated glucose transporter protein type 1 (GLUT1) deficiency syndrome: Impaired glucose transport into brain—a review. Eur. J. Pediatr. 161:295–304.

    Google Scholar 

  5. Nagamatsu, S., Kornhauser, J. M., Burant, C. F., Seino, S., Mayo, K. E., and Bell, G. I. 1992. Glucose transporter expression in brain. cDNA sequence of mouse GLUT3, the brain facilitative glucose transporter isoform, and identification of sites of expression by in situ hybridization. J. Biol. Chem. 267:467–472.

    Google Scholar 

  6. Brant, A. M., Jess, T. J., Milligan, G., Brown, C. M., and Gould, G. W. 1993. Immunological analysis of glucose transporters expressed in different regions of the rat brain and central nervous system. Biochem. Biophys. Res. Commun. 192:1297–1302.

    Google Scholar 

  7. Burcelin, R., Dolci, W., and Thorens, B. 2000. Glucose sensing by the hepatoportal sensor is GLUT2-dependent: In vivo analysis in GLUT2-null mice. Diabetes 49:1643–1648.

    Google Scholar 

  8. Leloup, C., Arluison, M., Lepetit, N., Cartier, N., Marfaing-Jallat, P., Ferre, P., and Penicaud, L. 1994. Glucose transporter 2 (GLUT 2): Expression in specific brain nuclei. Brain Res. 638:221–226.

    Google Scholar 

  9. Nualart, F., Godoy, A., and Reinicke, K. 1999. Expression of the hexose transporters GLUT1 and GLUT2 during the early development of the human brain. Brain Res. 824:97–104.

    Google Scholar 

  10. Leloup, C., Orosco, M., Serradas, P., Nicolaidis, S., and Penicaud, L. 1998. Specific inhibition of GLUT2 in arcuate nucleus by antisense oligonucleotides suppresses nervous control of insulin secretion. Brain Res. Mol. Brain Res. 57:275–280.

    Google Scholar 

  11. Waeber, G., Delplanque, J., Bonny, C., Mooser, V., Steinmann, M., Widmann, C., Maillard, A., Miklossy, J., Dina, C., Hani, E. H., Vionnet, N., Nicod, P., Boutin, P., and Froguel, P. 2000. The gene MAPK8IP1, encoding islet-brain-1, is a candidate for type 2 diabetes. Nat. Genet. 24:291–295.

    Google Scholar 

  12. Sankar, R., Thamotharan, S., Shin, D., Moley, K. H., and Devaskar, S. U. 2002. Insulin-responsive glucose transporters GLUT8 and GLUT4 are expressed in the developing mammalian brain. Brain Res. Mol. Brain. Res. 107:157–165.

    Google Scholar 

  13. Maekawa, F., Toyoda, Y., Torii, N., Miwa, I., Thompson, R. C., Foster, D. L., Tsukahara, S., Tsukamura, H., and Maeda, K. 2000. Localization of glucokinase-like immunoreactivity in the rat lower brain stem: For possible location of brain glucose-sensing mechanisms. Endocrinology 141:375–384.

    Google Scholar 

  14. Del Bigio, N. 1995. The ependyma: A protective barrier between brain and cerebrospinal fluid. Glia 14:1–13.

    Google Scholar 

  15. Bruni, J. E. 1998. Ependymal development, proliferation and functions: A literature review. Microsc. Res. Tech. 41:2–13.

    Google Scholar 

  16. Manthorpe, C. M., Wilkin, G. P., and Wilson, J. E. 1977. Purification of viable ciliated cuboidal ependymal cells from rat brain. Brain Res. 134:407–415.

    Google Scholar 

  17. Cathcart, R. S. and Worthington, W. C. 1964. Ciliary movement in the rat cerebral ventricles: Clearing action and direction of currents. J. Neuropath. Exp. Neurol. 23:609–618.

    Google Scholar 

  18. Roth, Y., Kimhi, Y., Edery, H., Aharonson, E., and Priel, Z. 1985. Ciliary motility in brain ventricular system and trachea of hamsters. Brain Res. 330:291–297.

    Google Scholar 

  19. Johansson, C. B., Momma, S., Clark, D. L., Risiling, M., Lendahl, U., and Frisén, J. 1999. Identification of a neural stem cell in the adult mammalian central nervous system. Cell 96:25–34.

    Google Scholar 

  20. Doetsch, F., Caillé, I, Lim, D. A., García-Verdugo, J. M., and Alvarez-Buylla, A. 1999. Subventricular zone astrocytes are neural stem cells in the adult mammalian brain. Cell 97:703–716.

    Google Scholar 

  21. Cataldo, A. M. and Broadwell, R. D. 1986. Cytochemical identification of cerebral glycogen and glucose-6-phosphatase activity under normal and experimental conditions: II. Choroid plexus and ependymal cells epithelia, endothelia and pericytes. J. Electron Microsc. Tech. 15:511–524.

    Google Scholar 

  22. Pfeiffer, B., Elmer, W., Roggendorf, W., Reinhart, P. H., and Hamprecht, B. 1990. Immunohistochemical demonstration of glycogen phosphorylase in rat brain slices. Histochemistry 94:73–80

    Google Scholar 

  23. Mikkelsen, J. D., Hay-Schmidt, A., and Larsen, P. J. 1997. Central innervation of the rat ependyma and subcommissural organ with special reference to ascending serotonergic projections from the raphe nuclei. J. Comp. Neurol. 384:556–558.

    Google Scholar 

  24. Simpson, K. L., Fisher, T. M., Waterhouse, B. D., and Lin, R. C. 1998. Projection patterns from the raphe nuclear complex to the ependymal wall of the ventricular system in the rat. J. Comp. Neurol. 399:61–72.

    Google Scholar 

  25. Prothmann, C., Wellard, J., Berger, J., Hamprecht, B., and Verleysdonk, S. 2001. Primary cultures as a model for studying ependymal functions: Glycogen metabolism in ependymal cells. Brain Res. 920:74–83.

    Google Scholar 

  26. Nguyen, T., Chin, W. C., O'Brien, J. A., Verdugo, P., and Berger, A. J. 2001. Intracellular pathways regulating ciliary beating of rat brain ependymal cells. J. Physiol. (Lond.) 531:131–140.

    Google Scholar 

  27. Eagle, H. 1959. Amino acid metabolism in mammalian cell cultures. Science 130:432–437.

    Google Scholar 

  28. Dulbecco, R. and Freeman, G. 1959. Plaque formation by the polyoma virus. Virology 8:396–397.

    Google Scholar 

  29. Suga, S., Nakao, K., Hosada, K., Mukoyama, M., Ogawa, Y., and Waymouth, C. 1959. Rapid proliferation of sublines of NCTC clone 929 (Strain L) mouse cells in a simple chemically defined medium (MB725/1). J. Natl. Cancer Inst. 22:1003–1017.

    Google Scholar 

  30. Miekka, S. I., Ingham, K. C., and Menache, D. 1982. Rapid methods for isolation of human plasma fibronectin. Thromb. Res. 27:1–14.

    Google Scholar 

  31. Weibel, M., Pettmann, B., Artault, J. C., Sensenbrenner, M., and Labourdette, G. 1986. Primary cultures of rat ependymal cells in serum-free defined medium. Dev. Brain Res. 25:199–209.

    Google Scholar 

  32. Hamprecht, B. and Löffler, F. 1985. Primary glial cultures as a model for studying hormone action. Methods Enzymol. 109:341–345.

    Google Scholar 

  33. Hamprecht, B. 1977. Structural, electrophysiological, biochemical, and pharmacological properties of neuroblastoma × glioma cell hybrids in cell culture. Int. Rev. Cytol. 49:99–170.

    Google Scholar 

  34. Hamprecht, B., Glaser, T., Reiser, G., Bayer, E., and Propst, F. 1985. Culture and characteristics of hormone-responsive neuroblastoma × glioma hybrid cells. Methods Enzymol. 109:316–340.

    Google Scholar 

  35. Burnette, W. N. 1981. 'Western blotting': Electrophoretic transfer of proteins from sodium dodecylsulfate-polyacrylamide gels to unmodified nitrocellulose and radiographic detection with antibody and radioiodinated protein A. Anal. Biochem. 112:195–203.

    Google Scholar 

  36. Bradford, M. M. 1974. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal. Biochem. 72:248–254.

    Google Scholar 

  37. Park, Y. C., Lee, C. H., Kang, H. S., Chung, H. T., and Kim, H. D. 1997. Wortmannin, a specific inhibitor of phosphatidylinositon-3-kinase, enhances LPS-induced NO production from murine peritoneal macrophages. Biochem. Biophys. Res. Commun. 240:692–696.

    Google Scholar 

  38. Clarke, D. W., Boyd, F. T. Jr., Kappy, M. S., and Raizada, M. K. 1984. Insulin binds to specific receptors and stimulates 2-deoxy-D-glucose uptake in cultured glial cells from rat brain. J. Biol. Chem. 259:11672–11675.

    Google Scholar 

  39. Hara, M., Matsuda, Y., Hirai, K., Okumura, N., and Nakagawa, H. 1989. Characteristics of glucose transport in neuronal cells and astrocytes from rat brain in primary culture. J. Neurochem. 52:902–908.

    Google Scholar 

  40. Hamai, M., Minokoshi, Y., and Shimazu, T. 1999. L-Glutamate and insulin enhance glycogen synthesis in cultured astrocytes from the rat brain through different intracellular mechanisms. J. Neurochem. 73:400–407.

    Google Scholar 

  41. Marshall, S., Bacote, V., and Traxinger, R. R. 1991. Discovery of a metabolic pathway mediating glucose-induced desensitization of the glucose transport system: Role of hexosamine biosynthesis in the induction of insulin resistance. J. Biol. Chem. 266:4706–4712.

    Google Scholar 

  42. Burant, C. F. and Bell, G. I. 1992. Mammalian facilitative glucose transporters: Evidence for similar substrate recognition sites in functionally monomeric proteins. Biochemistry 31:10414–10420.

    Google Scholar 

  43. Sato, S., Nishimura, H., Clark, A. E., Kozka, I. J., Vannucci, S. J., Simpson, I. A., Quon, M. J., Cushman, S. W., and Holman, G. D. 1993. Use of bismannose photolabel to elucidate insulin-regulated GLUT4 subcellular trafficking kinetics in rat adipose cells: Evidence that exocytosis is a critical site of hormone action. J. Biol. Chem. 268:17820–17829.

    Google Scholar 

  44. Yang, J. and Holman, G. D. 1993. Comparison of GLUT4 and GLUT1 subcellular trafficking in basal and insulin-stimulated 3T3-L1 cells. J. Biol. Chem. 268:4600–4603.

    Google Scholar 

  45. Okada, T., Kawano, Y., Sakakibara, T., Hazeki, O., and Ui, M. 1994. Essential role of phosphatidylinositol 3-kinase in insulin-induced glucose transport and antilipolysis in rat adipocytes: Studies with a selective inhibitor wortmannin. J. Biol. Chem. 269:3568–3573.

    Google Scholar 

  46. Maher, F. 1995. Immunolocalization of GLUT1 and GLUT3 glucose transporters in primary cultured neurons and glia. J. Neurosci. Res. 42:459–469.

    Google Scholar 

  47. Tomas, M., Fornas, E., Megias, L., Duran, J. M., Portoles, M., Guerri, C., Egea, G., and Renau-Piqueras, J. 2002. Ethanol impairs monosaccharide uptake and glycosylation in cultured rat astrocytes. J. Neurochem. 83:601–612.

    Google Scholar 

  48. Cummins, C. J., Glover, R. A., and Sellinger, O. Z. 1979. Neuronal cues regulate uptake in cultured astrocytes. Brain Res. 170:190–193.

    Google Scholar 

  49. Lisinski, I., Schurmann, A., Joost, H. G., Cushman, S. W., and Al-Hasani, H. 2001. Targeting of GLUT6 (formerly GLUT9) and GLUT8 in rat adipose cells. Biochem. J. 358:517–522.

    Google Scholar 

  50. Marshall, S. and Olefsky, J. M. 1980. Effects of insulin incubation on insulin binding, glucose transport, and insulin degradation by isolated rat adipocytes: Evidence for hormone-induced desensitization at the receptor and postreceptor level. J. Clin. Invest. 66:763–772.

    Google Scholar 

  51. Ercolani, L., Lin, H., and Ginsberg, B. H. 1985. Insulin-induced desenzitisation at the receptor and postreceptor level in mitogen-activated human T-lymphocytes. Diabetes 34:931–937.

    Google Scholar 

  52. Ballard, F. J., Wallace, J. C., Francis, G. L., Read, L. C., and Tomas, F. M. 1996. Des(1–3)IGF-I: a truncated form of insulin-like growth factor-I. Int. J. Biochem. Cell. Biol. 28:1085–1087.

    Google Scholar 

  53. Pulford, B. E. and Ishii, D. N. 2001. Uptake of circulating insulin-like growth factors (IGFs) into cerebrospinal fluid appears to be independent of the IGF receptors as well as IGF-binding proteins. Endocrinology 142:213–220.

    Google Scholar 

  54. Shen, S., Wertheimer, E., Sampson, S. R., and Tennenbaum, T. 2000. Characterization of glucose transport system in keratinocytes: Insulin and IGF-1 differentially affect specific transporters. J. Invest. Dermatol. 115:949–954.

    Google Scholar 

  55. Perez-Martin, M., Cifuentes, M., Grondona, J. M., Bermudez-Silva, F. J., Arrabal, P. M., Perez-Figares, J. M., Jimenez, A. J., Garcia-Segura, L. M., and Fernandez-Llebrez, P. 2003. Neurogenesis in explants from the walls of the lateral ventricle of adult bovine brain: Role of endogenous IGF-1 as a survival factor. Eur. J. Neurosci. 17:205–211.

    Google Scholar 

  56. Pierre, K., Pellerin, L., Debernardi, R., Riederer, B. M., and Magistretti, P. J. 2000. Cell-specific localization of monocarboxy-late transporters, MCT1 and MCT2, in the adult mouse brain revealed by double immunohistochemical labeling and confocal microscopy. Neuroscience 100:617–627.

    Google Scholar 

  57. Dringen, R., Gebhardt, R., and Hamprecht, B. 1993. Glycogen in astrocytes: Possible function as lactate supply for neighboring cells. Brain Res. 623:208–214.

    Google Scholar 

  58. Ibberson, M., Riederer, B. M., Uldry, M., Guhl, B., Roth, J., and Thorens, B. 2002. Immunolocalization of GLUTX1 in the testis and to specific brain areas and vasopressin-containing neurons. Endocrinology 143:276–284.

    Google Scholar 

  59. García, M. A., Millán, C., Balmaceda-Aguilera, C., Castro, T., Pastor, T., Montecinos, H., Reinicke, K., Zúñiga, F., Vera, J. C., Oñate, S. A., and Nualart, F. 2003. Hypothalamic ependymal-glial cells express the glucose transporter GLUT2 a protein involved in glucose sensing. J. Neurochem. (in press).

Download references

Author information

Affiliations

  1. Physiologisch-Chemisches Institut der Universität, Hoppe-Seyler-Str. 4, D-72076, Tübingen, Germany

    Stephan Verleysdonk, Wolfgang Hirschner, John Wellard & Bernd Hamprecht

  2. Aventis Behring GmbH, Emil-von-Behring-Str. 76, D-95041, Marburg, Germany

    Mirna Rapp

  3. Departamento Histología y Embriología, Facultad de Ciencias Biológicas, Universidad de Conceptión, Casilla 160-C, Conceptión, Chile

    Maria de los Angeles Garcia & Francisco Nualart

Authors
  1. Stephan Verleysdonk
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Wolfgang Hirschner
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. John Wellard
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Mirna Rapp
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Maria de los Angeles Garcia
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Francisco Nualart
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Bernd Hamprecht
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Stephan Verleysdonk.

Rights and permissions

Reprints and Permissions

About this article

Cite this article

Verleysdonk, S., Hirschner, W., Wellard, J. et al. Regulation by Insulin and Insulin-Like Growth Factor of 2-Deoxyglucose Uptake in Primary Ependymal Cell Cultures. Neurochem Res 29, 127–134 (2004). https://doi.org/10.1023/B:NERE.0000010441.08234.ca

Download citation

  • Issue Date:

  • DOI: https://doi.org/10.1023/B:NERE.0000010441.08234.ca

  • Ependymal cultures
  • glucose transport
  • insulin
  • IGF-1
  • kinetic parameters
  • GLUT1