Molecular Mechanisms of Action Induced by 5-HT3 Receptors in a Neuronal Cell Line and by 5-HT2 Receptors in a Glial Cell Line

  • G. Reiser
Chapter

Summary

The molecular mechanisms of action of serotonin were investigated in a neuronal cell line expressing 5-HT3 receptors (neuroblastoma × glioma hybrid cells) and in a glioma cell line with 5-HT2 receptors. In both cell lines, serotonin induces a transient rise of cytosolic Ca2+ activity. Ca2+ channel blockers (Ni2+ and La2+) suppress the Ca2+ response to serotonin in the neuronal cells but not in the glial cells. When internal Ca2+ stores are depleted and short-circuited by applying Ca2+ ionophores (ionomycin and A23187), serotonin still induces the normal Ca2+ response in the neuronal hybrid cells but not in the glioma cells. Thus, in the neuronal cell line cytosolic Ca2+ activity is enhanced through stimulation of Ca2+ entry into the cells from the extracellular environment via 5-HT3 receptors. The depolarization response caused by serotonin in these cells is due to activation of a cation conductance, and consequent entry of extracellular Ca2+. In the neuronal cell line, serotonin induces a rise of the cyclic GMP level, which depends on the rise of cytosolic Ca2+ activity. This conclusion is derived from the following findings: The serotonin-stimulated rise of cyclic GMP level is inhibited by i) reduced extracellular Ca2+ concentration (half-maximal stimulation at 0.3 mM Ca2+); ii) addition of inorganic (La3+, Mn2+, Co2+, Ni2+) or organic blockers (diltiazem, methoxyverapamil) of Ca2+ permeable channels; and iii) intracellular application of the Ca2+ chelator BAPTA. The suppression of the cyclic GMP effect of serotonin by the arginine analogues (NG-monomethyl-L-arginine, NG-nitro-L-arginine and canavanine) and by incubation in media containing oxyhemoglobin indicates that after stimulation with serotonin nitric oxide released from arginine acts as an intercellular stimulant of soluble guanylate cyclase. The rise of cytosolic Ca2+ activity appears to be a prerequisite for the formation of nitric oxide as an activator of guanylate cyclase. In the glial cell line, however, ketanserin-sensitive 5-HT2 receptors mainly cause liberation of Ca2+ from internal stores. In the glioma cells, Ca2+ released from internal stores opens Ca2+ -dependent K+ channels which results in the hyperpolarizing response.

Keywords

Nitric Oxide Glioma Cell Hybrid Cell Guanylate Cyclase Internal Store 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.
This is a preview of subscription content, log in to check access.

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. Ananth, U. S., Leli, U., and Hauser, G. (1987). Stimulation of phosphoinositide hydrolysis by serotonin in C6 glioma cells. J. Neurochem. 48: 253–261.CrossRefGoogle Scholar
  2. Christian, C. N., Nelson, P. G., Bullock, P., Mullinax, D., and Nirenberg, M. (1978). Pharmacological responses of cells of a neuroblastoma x glioma hybrid clone and modulation of synapses between hybrid cells and mouse myotubes. Brain Res. 147: 261–276.CrossRefGoogle Scholar
  3. Conn, P. J., and Sanders-Bush, E. (1985). Serotonin-stimulated phosphoinositide turnover: mediation by the S2 binding site in rat cerebral cortex but not in subcortical regions. J. Pharmacol. Exp. Ther. 234: 195–203.Google Scholar
  4. Donié F., and Reiser G. (1989). A novel, specific binding protein assay for quantitation of intracellular inositol 1,3,4,5-tetrakisphosphate (InsP4) using a high-affinity InsP4 receptor from cerebellum. FEBS Lett. 254: 155–158.CrossRefGoogle Scholar
  5. Fozard, J. R. (1984). MDL 72222: a potent and highly selective antagonist at neuronal 5-hydroxytryptamine receptors. Naunyn Schmiedeberg’s Arch. Pharmacol. 326: 36–44.CrossRefGoogle Scholar
  6. Furchgott, R. F., and Vanhoutte, P. M. (1989). Endothelium-derived relaxing and contracting factors. FASEB J. 3: 2007–2018.Google Scholar
  7. Grynkiewicz, G., Poenie, M., and Tsien, R. Y. (1985). A new generation of Cat+ indicators with greatly improved fluorescence properties. J. Biol. Chem. 260: 3440–3450.Google Scholar
  8. Gundersen, C. B., Miledi, R., and Parker, I. (1984). Messenger RNA from human brain induces drug-and voltage-operated channels in Xenopus oocytes. Nature (Lond.) 308: 421–424.CrossRefGoogle Scholar
  9. Hamprecht, B., Glaser, T., Reiser, G., Bayer, E., and Propst, F. (1985). Culture and characteristics of hormone-responsive neuroblastoma x glioma hybrid cells. Meth. Enzymol. 109: 316–341.CrossRefGoogle Scholar
  10. Heumann, R., Reiser, G., van Calker, D., and Hamprecht, B. (1982). Polyploid rat glioma cells: production, oscillations of membrane potential and response to neurohormones. Exp. Cell Res. 139: 117–126.CrossRefGoogle Scholar
  11. Hoyer, D., and Neijt, H. C. (1987). Identification of serotonin 5-HT3 recognition sites by radioligand binding in NG108–15 neuroblastoma-glioma cells. Eur. J. Pharmacol. 142: 291–292.CrossRefGoogle Scholar
  12. Jankowsky, A., Labarca, R., and Paul, S. A. (1984). Characterization of neurotransmitter receptor mediated phosphatidylinositol hydrolysis in the rat hippocampus. Life Sci. 35: 1953–1961.CrossRefGoogle Scholar
  13. Kilpatrick, G. J., Jones, B. J., and Tyers, M. B. (1987). Identification and distribution of 5-HT3 receptors in rat brain using radioligand binding. Nature (Lond.) 330: 746–748.CrossRefGoogle Scholar
  14. Lambert, J. J., Peters, J. A., Hales, T. G., and Dempster, J. (1989). The properties of 5-HT3 receptors in clonal cell lines studied by patch-clamp techniques. Br. J. Pharmacol. 97: 27–40.Google Scholar
  15. Leysen, J. E., Niemegeer, C. J. E., Van Neuten, J. M., and Laduron, P. M. (1982). [3H]Ketanserin (R41 468), as selective 3H-ligand for serotonin2 receptor binding sites: binding properties, brain distribution and functional role. Mol. Pharmacol. 21: 304–314.Google Scholar
  16. Lübbert, H., Hoffman, B. J., Snutch, T. P., van Dyke, T., Levine, A. J., Hartig, P. R., Lester, H. A., and Davidson, N. (1987). cDNA cloning of serotonin 5-HT,c receptor by electrophysiological assays of mRNA-injected Xenopus oocytes, Proc. Natl. Acad. Sci. USA. 84: 4332–4336.CrossRefGoogle Scholar
  17. Moncada, S., Palmer, R. M. J., and Higgs, E. A. (1989). Biosynthesis of nitric oxide from L-arginine. Biochem. Pharmacol. 38: 1709–1715.CrossRefGoogle Scholar
  18. Neijt, H. C., Plomb, J. J., and Vijverberg, H. P. M. (1989). Kinetics of the membrane current mediated by serotonin 5-HT3 receptors in cultured mouse neuroblastoma cells. J. Physiol. 411: 257–269.Google Scholar
  19. Ogura, A., and Amano, T. (1984). Serotonin-receptor coupled with membrane electrogenesis in a rat glioma clone. Brain Res. 297: 387–391.CrossRefGoogle Scholar
  20. Palmer, R. M. J., Ferrige, A. G., and Moncada, S. (1987). Nitric oxide release accounts for the biological activity of endothelium-derived relaxing factor. Nature (Lond.) 327: 524–526.CrossRefGoogle Scholar
  21. Peroutka, S. J. (1988). 5-Hydroxytryptamine receptor subtypes. Annu. Rev. Neurosci. 11: 45–60.CrossRefGoogle Scholar
  22. Peters, J. A., Malone, H. M., and Lambert, J. J. (1990). Antagonism of 5-HT3 receptor mediated currents in murine N1E-115 neuroblastoma cells by (+)-tubocurarine. Neurosci. Lett. 110: 107–112.CrossRefGoogle Scholar
  23. Pollock, W. K., Sage, S. O., and Rink, T. J. (1987). Stimulation of Cat+ efflux from fura-2-loaded platelets activated by thrombin or phorbol myristate acetate. FEBS Lett. 210: 132–136.CrossRefGoogle Scholar
  24. Reiser, G. (1990). Mechanism of stimulation of cyclic GMP level in a neuronal cell line mediated by serotonin (5-HT3) receptors: involvement of nitric oxide, arachidonic-acid metabolism and cytosolic Cat+. Eur. J. Biochem. 189: 547–552.CrossRefGoogle Scholar
  25. Reiser, G., Walter, U., and Hamprecht, B. (1984). Bradykinin regulates the level of guanosine 3’, 5’-cyclic monophosphate (cyclic GMP) in neural cell lines. Brain Res. 290: 367–371.CrossRefGoogle Scholar
  26. Reiser, G., Binmöller, F.-J., and Koch, R. (1988). Memantine (1-amino-3,5-dimethyladamantane) blocks the serotonin-induced depolarization response in a neuronal cell line. Brain Res. 443: 338–344.CrossRefGoogle Scholar
  27. Reiser, G., Donié, F., and Binmöller, F.-J. (1989). Serotonin regulates cytosolic Cat+ activity and membrane potential in a neuronal and in a glial cell line via 5-HT3- and 5-HT2-receptors by different mechanisms. J. Cell. Sci. 93: 545–555.Google Scholar
  28. Reiser, G., Binmöller, F.-J., and Donié, F. (1990a). Mechanisms for activation and subsequent removal of cytosolic Ca2± in bradykinin-stimulated neuronal and glial cell lines. Exp. Cell. Res. 186: 47–53.CrossRefGoogle Scholar
  29. Reiser G., Binmöller F.-J., Strong P. N., and Hamprecht B. (1990b). Activation of a K+ conductance by bradykinin and by inositol-1,4,5-trisphosphate in rat glioma cells: involvement of intracellular and extracellular Cat+. Brain Res. 506: 205–214.CrossRefGoogle Scholar
  30. Richardson, B. P., and Engel, G. (1986). The pharmacology and function of 5-HT3 receptors. Trends Neurosci. 9: 424–428.CrossRefGoogle Scholar
  31. Schmidt, H. H. H. W., Nau, H., Wittfoht, W., Gerlach, J., Prescher, K. E., Klein, M. M., Niroomand, F., and Böhme, E. (1988). Arginine is a physiological precursor of endothelium-derived nitric oxide. Eur. J. Pharmcol. 154: 213–216.CrossRefGoogle Scholar
  32. Waldman, S. A., and Murad, F. (1987). Cyclic GMP synthesis and function. Pharmacol. Rev. 39: 163–196.Google Scholar
  33. Yakel, J. L., and Jackson, M. B. (1988). 5-HT3 receptors mediate rapid responses in cultured hippocampus and a clonal cell line. Neuron 1: 615–621.CrossRefGoogle Scholar

Copyright information

© Birkhäuser Verlag Basel/Switzerland 1991

Authors and Affiliations

  • G. Reiser
    • 1
  1. 1.Physiologisch-chemisches InstitutUniversität TübingenTübingenGermany

Personalised recommendations