Abstract
Previous studies have demonstrated the presence of super-high affinity endothelin receptors with apparent Kd's on the order of pM in different brain tissues. This study was designed to characterize, in detail, the receptors present in SCP cells, a non-transformed sheep choroid plexus cell line. Competitive binding assays with receptor-selective ligands indicated the presence of at least three classes of binding sites: a conventional receptor of the ETA subtype with a Kd = 0.4 nM that mediates an increase in intracellular levels of inositol 1,4,5-trisphosphate (IP3) in response to ET-1 and two additional sites with much higher binding affinities. The latter two sites are not coupled to the common signal transduction pathways of IP3, cAMP and cGMP. Northern blot analysis confirmed the presence of only the ETA subtype mRNA in SCP cells. It remains to determined if the multiple binding sites are distinct gene products, multiple affinity states of a single receptor molecule or a result of cooperative association of one site with either the ligand or with other proteins.
Similar content being viewed by others
References
Inoue A, Yanagisawa M, Kimura S, Kasuya Y, Miyauchi T, Goto K, Masaki T: The human endothelin family: Three structurally and pharmacologically distinct isopeptides predicted by three separate genes. Proc Natl Acad Sci USA 86: 2864–2867, 1989
Yanagisawa M, Kurihara H, Kimura S, Tomobe Y, Kobayashi M, Mitsui Y, Yazaki Y, Goto K, Masaki T: A novel potent vasoconstrictor peptide produced by vascular endothelial cells. Nature 332: 411–415, 1988
Rubanyi GM, Polokoff MA: Endothelins: Molecular biology, biochemistry, pharmacology, physiology, and pathophysiology. Pharmacol Rev 46: 325–415, 1994
Giaid A, Gibson SJ, Ibrahim NBN, Legon S, Bloom SR, Yanagisawa M, Masaki T, Varndell JM, Polak IM: Endothelin-1, an endothelium-derived peptide is expressed in neurons of the human spinal cord and dorsal root ganglia. Proc Natl Acad Sci USA 86: 7634–7638, 1989
Giaid A, Gibson SJ, Herrero MY, Gentelman S, Legon S, Yanagisawa M, Masaki T, Ibrahim NBN, Roberts GW, Rossi ML, Polak JM: Topographical localization of endothelin mRNA and peptide immunoreactivity in neurones of the human brain. Histochemistry 95: 303–314 1991
Lee ME, de la Monte S, Bloch KD, Quartermous T: Expression of the potent vasoconstrictor endothelin in the human central nervous system. J Clin Invest 86: 141–147, 1990
MacCumber MW, Ross CA, Snyder SH: Endothelin in brain: Receptors, mitogenesis, and biosynthesis in glial cells. Proc Natl Acad Sci USA 87: 2359–2363, 1990
Matsumoto H, Suzuki N, Onda H, Fujino M: Abundance of endothelin-3 in rat intestine, pituitary gland and brain. Biochem Biophys Res Commun 164: 74–80, 1989
Kuwaki T, Cao W-H, Kumada M: Endothelin in the brain and its effect on central control of the circulation and other functions. Jpn J Physiol 44: 1–18, 1994
Takayanagi R, Ohnaka K, Takasaki C, Ohashi M, Nawata H: Multiple subtypes of endothelin receptors in porcine tissues: Characterized by ligand binding, affinity labeling and regional distribution. Regul Peptides 32: 23–37, 1991
Sakurai T, Yanagisawa M, Takuwa Y, Miyazaki H, Kimura S, Goto K, Masaki T: Cloning of a cDNA encoding a non-isopeptide-selective subtype of the endothelin receptor. Nature 348: 732–735, 1990
Arai H, Hori S, Aramori I, Ohkubo H, Nakanishi S: Cloning and expression of a cDNA encoding an endothelin receptor. Nature 348: 730–732, 1990
Lin HY, Kaji EH, Winkel GK, Ives HE, Lodish HF: Cloning and functional expression of a vascular smooth muscle endothelin 1 receptor. Proc Natl Acad Sci USA 88: 3185–3189, 1991
Jones CR, Hiley CR, Pelton JT, Mohr M: Autoradiographic visualization of the binding sites for [125I]endothelin in rat and human brain. Neurosci Lett 97: 276–279, 1989
Kohzuki M, Chai SY, Paxinos G, Karavas A, Casley DJ, Johnston CI, Mendelsohn FAO: Localization and characterization of endothelin receptor binding sites in the rat brain visualized by in vitro autoradiography. Neurosci 42: 245–260, 1991
Fuxe K, Anggard E, Lundgren K, Cintra A, Agnati LF, Galron S, Vane J: Localization of [125I]endothelin-1 and [125III]endothelin-3 binding sites in the rat brain. Acta Physiol Scand 137: 563–564 1989
Hoyer D, Waeber C, Palacios JM: [125I]endothelin-1 binding sites: Autoradiographic studies in the brain and periphery of various species including humans. J Cardiovasc Pharmacol 13(Suppl. 5): S162-S165, 1989
Koseki C, Imai M, Hirata Y, Yanagisawa M, Masaki T: Autoradiographic distribution in rat tissues of binding sites for endothelin: A neuropeptide. Am J Physiol 256: R858-R866, 1989
Sokolovsky M, Ambar I, Galron R: A novel subtype of endothelin receptors. J Biol Chem 267: 20551–20554, 1992
Sokolovsky M, Shraga-Levine Z, Galron R: Ligand-specific stimulation/inhibition of cAMP formation by a novel endothelin receptor subtype. Biochemistry 33: 11417–11419, 1994
Shraga-Levine Z, Galron R, Sokolovsky M: Cyclic GMP formation in rat cerebellar slices is stimulated by endothelins via nitric oxide formation and by sarafotoxins via formation of carbon monoxide. Biochemistry 33: 14656–14659, 1994
Palmer S, Hughes KT, Lee DY, Wakelam MJO: Development of a novel ins(1,4,5)P3-specific binding assay. Its use to determine the intracellular concentration of ins(1,4,5)P3 in unstimulated and vasopressin-stimulated rat hepatocytes. Cell Signalling 1: 147–156, 1989
Angelova K, Puett D, Ergul A: Characterization of endothelin receptors on neuroblastoma-glioma hybrid cells. Abstracts of the 76th Annual Meeting of the Endocrine Society abstract no. 1167, 1994
Kanyicska B, Freeman ME: Characterization of endothelin receptors in the anterior pituitary gland. Am J Physiol 265: E601-E608, 1993
Kumar C, Mwangi V, Nuthulaganti P, Wu H-L, Pullen M, Brun K, Aiyar H, Morris RA, Naughton R, Nambi P: Cloning and characterization of a novel endothelin receptor from Xenopus heart. J Biol Chem 269: 13414–13420, 1994
Nambi P, Pullen M, Kumar C: Identification of a novel endothelin receptor in Xenopus laevis liver. Neuropeptides 26: 181–185, 1994
Elshourbagy NA, Lee JA, Korman DR, Nuthalanganti P, Sylvester DR, Dilella AG, Sutiphong JA, Kumar CS: Molecular cloning and characterization of the major endothelin receptor subtype in porcine cerebellum. Mol Pharmacol 41: 465–473, 1992
Fernandez-Durango R., de Juan JA, Zimman H, Moya FJ, Garcia de la Coba M, Fernandez-Cruz A: Identification of endothelin receptor subtype (ETB) in human cerebral cortex using subtype-selective ligands. J Neurochem 62: 1482–1488, 1994
Huggins JP, Pelton JT, Miller RC: The structure and specificity of endothelin receptors: their importance in physiology and medicine. Pharmac Ther 59: 55–123, 1993
Sakamoto A, Yanagisawa M, Sakurai T, Takuwa Y, Yanagisawa H, Masaki T: Cloning and functional expression of human cDNA for the ETB endothelin receptor. Biochem Biophys Res Commun 178: 656–663, 1991
Haendler B, Hechler U, Schleuning W-D: Molecular cloning of human endothelin (ET) receptors ETA and ETB. J Cardiovasc Pharm 20(Suppl.12): S1-S4, 1992
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Angelova, K., Fralish, G.B., Puett, D. et al. Identification of conventional and novel endothelin receptors in sheep choroid plexus cells. Mol Cell Biochem 159, 65–72 (1996). https://doi.org/10.1007/BF00226064
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00226064