TAILIEUCHUNG - Báo cáo Y học: The SK-N-MC cell line expresses an orexin binding site different from recombinant orexin 1-type receptor

Orexin A and B (also known as hypocretins), two recently discovered neuropeptides, play an important role in food intake, sleep/wake cycle and neuroendocrine functions. Orexins are endogenous ligands of two G-protein-coupled receptors, termed OX1 and OX2. This work presents the first short orexin A and B analogues, orexin A 23–33 and orexin B 18–28, with high affinity (119 ± 49 and 49 ± 23 nM) for OX1 receptors expressed on SK-N-MC cells and indicates the importance of the C-terminal part of the orexin peptides for this ligand–receptor interaction. However, these C-terminal fragments of orexin did not displace the 125 I-labelled. | Eur. J. Biochem. 269 1128-1135 2002 FEBS 2002 The SK-N-MC cell line expresses an orexin binding site different from recombinant orexin 1-type receptor Heike A. Wieland1 Richard M. Still2 3 Henri N. Doods1 Dirk Stenkamp1 Rudolf Hurnaus1 Barbel Lammle1 and Annette G. Beck-Sickinger2 Division of Preclinical Research Boehringer Ingelheim Pharma KG Biberach Germany 2Institute of Biochemistry University of Leipzig Germany 3Department of Applied Biosciences Swiss Federal Institute of Technology Zurich Switzerland Orexin A and B also known as hypocretins two recently discovered neuropeptides play an important role in food intake sleep wake cycle and neuroendocrine functions. Orexins are endogenous ligands of two G-protein-coupled receptors termed OX1 and OX2. This work presents the first short orexin A and B analogues orexin A 23-33 and orexin B 18-28 with high affinity 119 49 and 49 23nM for OX1 receptors expressed on SK-N-MC cells and indicates the importance of the C-terminal part of the orexin peptides for this ligand-receptor interaction. However these C-ter-minal fragments of orexin did not displace the 125I-labelled orexin B from the recombinant orexin 1 receptor stably expressed in Chinese hamster ovary cells. To examine the role of the shortened orexin A 23-33 in feeding its effects in mimicking or antagonizing the effects of orexin A were studied in rats after administration via the lateral hypothalamus. In contrast with orexin A which potently induced feeding up to 4 h after administration orexin A 23-33 neither induced feeding nor inhibited orexin A-induced feeding. Modafinil Vigil which was shown earlier to activate orexin neurons displayed binding neither to the orexin receptor expressed on SK-N-MC cells nor to the recombinant orexin 1 receptor which indicates that modafinil displays its antinarcoleptic action via another yet unknown mechanism. PCR and subsequent sequencing revealed expression of the full-length orexin 1 receptor mRNA in SK-N-MC and NT-2 .

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