Abstract
The opioid receptor-like1 receptor (ORL1), an orphan receptor whose human and murine complementary DNAs, has been characterized recently. ORL1 transcripts are particularly abundant in the central nervous system. We demonstrated that ORL1 expressed in human neuroblastoma SK-N-SH and SH-SY5Y cell lines by radioligand binding assay, reverse transcription polymerase chain reaction (RT-PCR) and Northern analysis in the present study. Stimulation with ORL1 specific agonist, nociceptin/orphanin FQ, increased [35S]GTP γ S binding to SK-N-SH cell membranes (EC50 = 14 ± 0.45 nM), and attenuated, forskolin-stimulated accumulation of cellular cAMP (EC50 = 0.80 ± 0.45 nM), indicative that activation of ORL1 activates G proteins and inhibits adenylyl cyclase. Activation of ORL1 receptor was also accessed using CHO:hORL1 cell line by microphysiometer. Treatment of nociceptin/orphanin FQ increased extracellular acidification rate significantly.
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References
Reisine T . Opiate receptors. Neuropharmacology 1995; 34:463–72.
Mollereau C, Parmentier M, Mailleux P et al. ORL1, a novel member of the opioid receptor family Cloning, functional expression and localization. FFBS Letters 1994; 341:33–8.
Fukuda K, Kato S, Mori K, et al. cDNA cloning and regional distribution of a novel member of the opioid receptor family. FEBS Letters 1994; 343:42–6.
Chen Y, Fan Y, Liu J, et al. Molecular cloning, tissue distribution and chromosomal localization of a novel member of the opioid receptor gene family. FEBS Letters 1994; 347:279–83.
Bunzow JR, Saez C, Mortrud M, et al. Molecular cloning and tissue distribution of a putative member of the rat opioid receptor gene family that is not a μ, δ or κ opioid receptor type. FEBS Letters 1994; 347:284–8.
Wang JB, Johnson PS, Imai Y . DNA cloning of an orphan opiate receptor gene family member and its splice variant. FEBS Letters 1994; 348:75–9.
Wick M J, Minnerah SR, Lin X, Elde R, Law PY, Loh HH . Isolation of a novel cDNA encoding a putative memberane receptor with high homology to the cloned μ, δ or κ opioid receptors. Mol Brain Res 1994; 27:37–44.
Lachowicz JE, Shen Y, Monsma F J, Jr, Sibley DR . Molecular cloning of a novel G proteincoupled receptor related to the opioid receptor family. J Neurochem 1995; 64:34–40.
Meunier J-C, Mollereau C, Toll L . Isolation and structure of the endogenous agonist of opioid receptor-like ORL1 receptor. Nature 1995; 377:532–5.
Reinscheid R . Nothacker H-P ., Bourson A, et al. Orphanin FQ: a neuropeptide that activates an opioid-like G protein-coupled receptor. Science 1995; 270:792–4.
Sim L J, Xiao R, Childers SR . Indentification of opioid receptor-like (ORL1) peptide-sitmulated [35S] GTP gamma S binding in rat brain. Neuro Report 1996; 7:729–33.
Vaughan CW, Christie MJ . Increase by the ORL1 receptor (opioid receptor-like1) ligand, nociceptin, of inwardly rectifying K conductance in dorsal raphe nucleus neurones. Brit J Pharmacol 1996:117:1609–11.
Hochhaus G, Yu VC, Sadee W, Delta opioid receptors in human neuroblastoma cell lines. Brain Res 1986; 382:327–31.
Church, Gilbert, Genomic sequencing. PNAS 1984; 81:1991–5.
Velu T J, Beguinot L, Vas WC, Zhang K, Pastan I, Lowy DR . Retroviruses expressing different levels of the normal epidermal growth factor receptor: biological properties and new bioassay. J Cell-Biochem 1989; 39:153–66.
Pei G, Kieffer BL, Lefkowitz RJ, Neil JF . Agonist-dependent phosphorylation of the mouse δ-opioid receptor: involvement of G protein-coupled receptor kinasas but not protein kinase C. Mol Pharmacol 1995; 48:173–7.
Cai YC, Zhang Y, Wu Y, Pei G . δ opioid receptor in neuronal cell undergoes acute and homologous desensitization. Biochem Biophys Res Comm 1996; 219:342–7.
Anton B, Fein J, To T, Li X, Silberstein L, Evens C . Immunohistochemical localization of ORL1 in the central nervous system of the rat. J Comp Neurol 1996; 368:229–51.
Houtani T, Nishi M, Takeshima H, Nukada T, Sugimoto T . Structure and regional distribution of nociceptin/orphanin FQ precursor. Biochem Biophys Res Comm 1996; 219:714–9.
Connor M, Yeo A, Henderson G . The effect of nociceptin on Ca2+ channel current and intracellure Ca2+ in the SH-SY5Y human neurolastoma cell line. Br J Pharmacol 1996; 118:205–7.
Acknowledgements
This research was supported by research grants from Chinese Academy of Sciences, German Max-Planck Society, Shanghai Educational Development Foundation and Natural Science Foundation of China (39630130 and 39625010). The authors thank Guo-Huang FAN, Li-Zhen JIANG, Ye ZHU, and Yan-Ping WANG for their help.
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*Dedicated to Professor Lu Ji SHI for his 80th birthday
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Wu, Y., Pu, L., Ling, K. et al. Molecular characterization and functional expression of opioid receptor-like1 receptor. Cell Res 7, 69–77 (1997). https://doi.org/10.1038/cr.1997.8
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DOI: https://doi.org/10.1038/cr.1997.8
