Orexins/Hypocretins Acting at Gi Protein-Coupled OX2 Receptors Inhibit Cyclic AMP Synthesis in the Primary Neuronal Cultures

• Published in: Journal of molecular neuroscience Vol. 46; no. 1; pp. 10 - 17
• Main Authors: Urbańska, Anna, Sokołowska, Paulina, Woldan-Tambor, Agata, Biegańska, Kaja, Brix, Britta, Jöhren, Olaf, Namiecińska, Magdalena, Zawilska, Jolanta Barbara
• Format: Journal Article
• Language: English
• Published: New York Humana Press Inc 2012
• Subjects: Biomedical and Life Sciences; Biomedicine; Cell Biology; Neurochemistry; Neurology; Neurosciences; Proteomics; Cerebral cortex; PACAP; Rat; Orexin receptors; Cyclic AMP; Neuronal cell culture; VIP; Orexin; Hypocretin
• ISSN: 0895-8696; 1559-1166
• DOI: 10.1007/s12031-011-9526-2

Orexins A and B are newly discovered neuropeptides with pleiotropic activity. They signal through two G protein-coupled receptors: OX 1 and OX 2 . In this study, we examined the expression of orexin receptors and effects of the receptors’ activation on cyclic AMP formation in the primary neuronal cell cultures from rat cerebral cortex. Both types of orexin receptors were expressed in rat cortical neurons; the level of OX 2 R was markedly higher compared to OX 1 R. Orexin A (an agonist of OX 1 R and OX 2 R) and [Ala 11 - D -Leu 15 ]orexin B (a selective agonist of OX 2 R) did not affect basal cyclic AMP formation in the primary neuronal cell cultures. Both peptides (0.001–1 μM) inhibited, in a concentration-dependent manner and IC 50 values in low nanomolar range, the increase in the nucleotide production evoked by forskolin (1 μM; a direct activator of adenylyl cyclase), pituitary adenylate cyclase-activating polypeptide (PACAP27; 0.1 μM), and vasoactive intestinal peptide (VIP; 3 μM). Effects of orexin A on forskolin-, PACAP27-, and VIP-stimulated cyclic AMP synthesis were blocked by TCS OX2 29 (a selective antagonist of OX 2 R), and unaffected by SB 408124 (a selective antagonist of OX 1 R). Pretreatment of neuronal cell cultures with pertussis toxin (PTX) abolished the inhibitory action of orexin A on forskolin- and PACAP-stimulated cyclic AMP accumulation. It is suggested that in cultured rat cortical neurons orexins, acting at OX 2 receptors coupled to PTX-sensitive G i protein, inhibit cyclic AMP synthesis.