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.
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References
Ananth, U. S., Leli, U., and Hauser, G. (1987). Stimulation of phosphoinositide hydrolysis by serotonin in C6 glioma cells. J. Neurochem. 48: 253–261.
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.
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.
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.
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.
Furchgott, R. F., and Vanhoutte, P. M. (1989). Endothelium-derived relaxing and contracting factors. FASEB J. 3: 2007–2018.
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.
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.
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.
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.
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.
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.
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.
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.
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.
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.
Moncada, S., Palmer, R. M. J., and Higgs, E. A. (1989). Biosynthesis of nitric oxide from L-arginine. Biochem. Pharmacol. 38: 1709–1715.
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.
Ogura, A., and Amano, T. (1984). Serotonin-receptor coupled with membrane electrogenesis in a rat glioma clone. Brain Res. 297: 387–391.
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.
Peroutka, S. J. (1988). 5-Hydroxytryptamine receptor subtypes. Annu. Rev. Neurosci. 11: 45–60.
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.
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.
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.
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.
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.
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.
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.
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.
Richardson, B. P., and Engel, G. (1986). The pharmacology and function of 5-HT3 receptors. Trends Neurosci. 9: 424–428.
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.
Waldman, S. A., and Murad, F. (1987). Cyclic GMP synthesis and function. Pharmacol. Rev. 39: 163–196.
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.
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Reiser, G. (1991). Molecular Mechanisms of Action Induced by 5-HT3 Receptors in a Neuronal Cell Line and by 5-HT2 Receptors in a Glial Cell Line. In: Fozard, J.R., Saxena, P.R. (eds) Serotonin: Molecular Biology, Receptors and Functional Effects. Birkhäuser Basel. https://doi.org/10.1007/978-3-0348-7259-1_7
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DOI: https://doi.org/10.1007/978-3-0348-7259-1_7
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