Skip to main content
Log in

Expression of the extraneuronal monoamine transporter (uptake2) in human glioma cells

  • Original Article
  • Published:
Naunyn-Schmiedeberg's Archives of Pharmacology Aims and scope Submit manuscript

Abstract

Tritiated methylphenylpyridinium ([3H]MPP+), a substrate of the neuronal and extraneuronal noradrenaline transporter (uptake1 and uptake2, respectively) and of the organic cation transporter (OCT1), was used to characterize the amine transport system of the established human glioma cell line SK-MG-1.

Uptake of [3H]MPP+ (25 nM) into SK-MG-1 cells increased linearly with time for up to 15 min. Selective uptake1 inhibitors (e.g. (+)oxaprotiline) or omission of Na+ or Cl ions did not affect [3H]MPP+ uptake, whereas uptake2 inhibitors such as O-methyl-isoprenaline (OMI) or corticosterone as well as depolarizing concentrations of K+ or Ba2+ strongly reduced [3H]MPP+ uptake. Initial rates of OMI(100 μM)-sensitive [3H]MPP+ uptake were saturable, with a Km of about 17 μM and a maximal rate of about 50 pmol/ (min × mg protein). IC50 (or Ki) values for inhibition of [3H]MPP+ uptake by substrates and inhibitors of uptake2 or OCTI were highly significantly correlated with published IC50 values for inhibition of uptake2 but not with corresponding values for inhibition of OCT1.

The results presented here clearly demonstrate that human glioma cells express an uptake2 transporter. Thus, glial cells in the human central nervous system endowed with this transporter are likely to contribute to the inactivation of neuronally released noradrenaline.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  • Amara SG, Kuhar MJ (1993) Neurotransmitter transporters: recent progress. Annu Rev Neurosci 16:73–93

    Google Scholar 

  • Amundson RH, Goderie SK, Kimelberg HK (1992) Uptake of [3H]serotonin and [3H]glutamate by primary astrocyte cultures. II. Differences in cultures prepared from different brain regions. Glia 6:9–18

    Google Scholar 

  • Bönisch H (1980) Extraneuronal transport of catecholamines. Pharmacology 21:93–108

    Google Scholar 

  • Bönisch H, Brüss M (1994) The noradrenaline transporter of the neuronal plasma membrane. Ann New York Acad Sci 733:193–202

    Google Scholar 

  • Bönisch H, Bryan LJ, Henseling M, O'Donnell SR, Stockmann P, Trendelenburg U (1985) The effect of various ions on uptake2 of catecholamines. Naunyn-Schmiedeberg's Arch Pharmacol 328:407–416

    Google Scholar 

  • Callingham BA, Burgen ASV (1966) The uptake of isoprenaline and noradrenaline by the perfused rat heart. Mol Pharmacol 2:37–42

    Google Scholar 

  • Cheng Y, Prusoff WH (1973) Relationship between the inhibition contant (KI) and the concentration which causes 50% inhibition of an enzymatic reaction. Biochem Pharmacol 22:403–411

    Google Scholar 

  • Graefe K-H, Bönisch H (1988) The transport of amines across the axonal membrane of noradrenergic and dopaminergic neurones. In: Trendelenburg U, Weiner N (eds) Handbook of Experimental Pharmacology, Vol 90/1. Springer, Berlin Heidelberg New York, pp 193–245

    Google Scholar 

  • Grohmann M, Trendelenburg U (1984) The substrate specificity of uptake2 in the rat heart. Naunyn-Schmiedeberg's Arch Pharmacol 328:164–173

    Google Scholar 

  • Gründemann D, Gorboulev V, Gambaryan S, Veyhl M, Koepsell H (1994) Drug excretion mediated by a new prototype of polyspecific transporter. Nature 372:549–552

    Google Scholar 

  • Hansson E (1984) Enzyme activities of monoamine oxidase, catechol-O-methyltransferase and gamma- aminobutyric acid transaminase in primary astroglial cultures and adult brain from different brain regions. Neurochem Res 9:45–57

    Google Scholar 

  • Hansson E (1988) Astroglia from defined brain regions as studied with primary cultures. Progr Neurobiol 30:369–397

    Google Scholar 

  • Hansson E, Rönnbäck L (1992) Adrenergic receptor regulation of amino acid neurotransmitter uptake in astrocytes. Brain Res Bull 29:297–301

    Google Scholar 

  • Hendley ED (1976) The mechanism of extraneuronal transport of catecholamines in the central nervous system. In: Paton DM (ed) The mechanism of neuronal and extraneuronal transport of catecholamines. Raven Press, New York, pp 313–324

    Google Scholar 

  • Hendley ED, Taylor KM, Snyder SH (1970) 3H-Normetanephrine uptake in rat brain slices. Relationship to extraneuronal accumulation of norepinephrine. Eur J Pharmacol 12:167–179

    Google Scholar 

  • Hösli E, Hösli L (1993) Receptors for neurotransmitters on astrocytes in the mammalian central nervous system. Prog Neurobiol 40:477–506

    Google Scholar 

  • Iversen LL (1967) The uptake and storage of noradrenaline in sympathetic nerves. Cambridge University Press, Cambridge, England

    Google Scholar 

  • Javitch JA, D' Amato RJ, Strittmatter SM, Snyder SH (1985) Parkinsonism-inducing neurotoxin, N-methyl-4-phenyl-1,2,3,6-tetrahydropyridine: Uptake of the metabolite N-methyl-4-phenylpyridine by dopamine neurons explains selective toxicity. Proc Natl Acad Sci USA 82:2173–2177

    Google Scholar 

  • Jennings MT, Jennings VDI, Asadourian LJH, Ebrahim SAD, Klein CE, Old LJ (1989) Antigenic phenotypes of cultured malignant astrocytomas: identification of lineage-independent and putative tumor-restricted antigenic expression. J Neurol Sci 89:79–92

    Google Scholar 

  • Katz DM, Kimelberg HK (1985) Kinetics and autoradiography of high affinity uptake of serotonin by primary astrocyte cultures. J Neurosci 5:1901–1908

    Google Scholar 

  • Kimelberg HK (1986) Catecholamine and serotonin uptake in astrocytes. In: Fedoroff S, Vernadakis A (eds) Astrocytes: Biochemistry, physiology and pharmacology of astrocytes, vol 2. Academic Press, Orlando, pp 107–131

    Google Scholar 

  • Kimelberg HK, Pelton EW (1983) High-affinity uptake of [3H]norepinephrine by primary astrocyte cultures and its inhibition by tricyclic antidepressants. J Neurochem 40:1265–1270

    Google Scholar 

  • Kimelberg HK, Goderie SK, Conley PA, Higman S, Goldschmidt R (1992) Uptake of [3H]serotonin and [3H]glutamate by primary astrocyte cultures. I. Effects of different sera and time in culture. Glia 6:1–8

    Google Scholar 

  • Köster G (1995) Glial α2-receptors probably inhibit the high-affinity uptake of noradrenaline into astrocytes in the rat brain in vivo. Neurochem Res 20:291–297

    Google Scholar 

  • Lingen B, Brüss M, Bönisch H (1994) Cloning and expression of the bovine sodium- and chloride-dependent noradrenaline transporter. FEBS Lett 342:235–238

    Google Scholar 

  • Lowry OH, Rosebrough NJ, Farr AL, Randall RJ (1951) Protein measurement with the Folin phenol reagent. J Biol Chem 193:265–275

    Google Scholar 

  • Mack F, Bönisch H (1979) Dissociation constants and lipophilicity of catecholamines and related compounds. Naunyn-Schmiedeberg's Arch Pharmacol 310:1–9

    Google Scholar 

  • Pacholczyk T, Blakely RD, Amara SG (1991) Expression cloning of a cocaine- and antidepressant-sensitive human noradrenaline transporter. Nature 350:350–354

    Google Scholar 

  • Paterson IA, Hertz L (1989) Sodium-independent transport of noradrenaline in mouse and rat astrocytes in primary culture. J Neurosci Res 23:71–77

    Google Scholar 

  • Pelton EW, Kimelberg HK, Shipherd SV, Bourke RS (1981) Dopamine and norepinephrine uptake and metabolism by astroglial cells in culture. Life Sci 28:1655–1663

    Google Scholar 

  • Raisman R, Sette M, Pimoule C, Briley M, Langer SZ (1982) High-affinity 3H-desipramine binding in the peripheral and central nervous system: a specific site associated with the neuronal uptake of noradrenaline. Eur J Pharmacol 78:345–352

    Google Scholar 

  • Russ H, Gliese M, Sonna J, Schdmig E (1992) The extraneuronal transport mechanism for noradrenaline (uptake2) avidly transports 1-methyl-4-phenylpyridinium (MPP+). Naunyn-Schmiedeberg's Arch Pharmacol 346:158–165

    Google Scholar 

  • Schömig E, Schönfeld CL (1990) Extraneuronal noradrenaline transport (uptake2) in a human cell line (CAKI-1). NaunynSchmiedeberg's Arch Pharmacol 341:404–410

    Google Scholar 

  • Semenoff D, Kimelberg HK (1985) Autoradiography of high affinity uptake of catecholamines by primary astrocyte cultures. Brain Res 348:125–136

    Google Scholar 

  • Skalski V, Feindel W, Panasci LC (1990) Transport of amino acid amide sarcosinamide and sarcosinamide chloroethylnitrosourea in human glioma SK-MG-1 cells. Cancer Res 50:3062–3066

    Google Scholar 

  • Sontheimer H (1992) Astrocytes, as well as neurons, express a diversity of ion channels. Can J Physiol Pharmacol 70:S223-S238

    Google Scholar 

  • Staudt K, Russ H, Gliese M, Schdmig E (1993) The extraneuronal noradrenaline carrier (uptake2) exists in the human central nervous system (CNS). Naunyn-Schmiedeberg's Arch Pharmacol 347 [Suppl]:R118

    Google Scholar 

  • Trendelenburg U (1988) The extraneuronal uptake and metabolism of catecholamines. In Trendelenburg U, Weiner N (eds) Handbook of Experimental Pharmacology, Vol 90/1. Springer, Berlin Heidelberg New York, pp 279–319

    Google Scholar 

  • Tsai MJ, Lee EHY (1994) Differences in the disposition and toxicity of 1-methyl-4-phenylpyridinium in cultured rat and mouse astrocytes. Glia 12:329–335

    Google Scholar 

  • Ullrich KJ, Rumrich G, Neiteler K, Fritzsch G (1992) Contraluminal transport of organic cations in the proximal tubule of the rat kidney. II. Specificity: anilines, phenylalkylamines (catecholamines), heterocyclic compounds (pyridines,quinolines, acridines). Plügers Arch 420:29–38

    Google Scholar 

  • Waldmeier PC, Baumann PA, Hauser K, Maitre L, Storm A (1982) Oxaprotiline, a noradrenaline uptake inhibitor with an active and an inactive enantiomer. Biochem Pharmacol 31:2169–2176

    Google Scholar 

  • Wall SC, Gu H, Rudnick G (1995) Biogenic amine flux mediated by cloned transporters stably expressed in cultured cell lines: amphetamine specificity for inhibition and efflux. Mol Pharmacol 47:544–550

    Google Scholar 

  • Wilson VG, Grohmann M, Trendelenburg U (1988) The uptake and O-methylation of 3H-(±)-isoprenaline in rat cerebral cortex slices. Naunyn-Schmiedeberg's Arch Pharmacol 337:397–405

    Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Rights and permissions

Reprints and permissions

About this article

Cite this article

Streich, S., Miss, M. & Bönisch, H. Expression of the extraneuronal monoamine transporter (uptake2) in human glioma cells. Naunyn-Schmiedeberg's Arch Pharmacol 353, 328–333 (1996). https://doi.org/10.1007/BF00168636

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00168636

Key words

Navigation