title: Integrins regulate opioid receptor signaling in trigeminal ganglion neurons.
abstract: The binding of integrins to the extracellular matrix results in focal organization of the cytoskeleton and the genesis of intracellular signals that regulate vital neuronal functions. Recent evidence suggests that integrins modulate G-protein-coupled receptor (GPCR) signaling in hippocampal neurons. In this study we evaluated the hypothesis that integrins regulate the mu opioid receptor in rat trigeminal ganglion neurons. For these studies, primary cultures of adult rat trigeminal ganglion neurons were used to demonstrate the colocalization of beta1 and beta3 integrins with mu opioid receptor in caveolin-1-rich membrane fractions, and at focal adhesions sites generated by integrin ligand binding. Furthermore, we show that the mu opioid receptor agonist, DAMGO ([D-Ala(2),N-MePhe(4),Gly-ol(5)]enkephalin), inhibits cyclic AMP (cAMP) accumulation in response to prostaglandin E2 (PGE(2)) stimulation in bradykinin-primed, but not unprimed, cultured trigeminal ganglia neurons. Application of soluble GRGDS (Gly-Arg-Gly-Asp-Ser) peptides that bind specific integrins (i.e. RGD-binding integrins) completely abolished the DAMGO effect in bradykinin-primed trigeminal ganglia neurons, but did not alter bradykinin-mediated hydrolysis of phosphatidylinositol. Likewise, monospecific anti-beta1 and anti-beta3 integrin subunit antibodies blocked this DAMGO effect in bradykinin-primed trigeminal ganglia neurons. Indeed, application of anti-beta1 integrin subunit actually reversed DAMGO signaling, resulting in increased cAMP accumulation in these cells. This suggests that the relative amounts of specific activated integrins at focal adhesions may govern signaling by the mu opioid receptor, perhaps by altering interactions with G proteins (e.g. Galphai vs. Galphas). Collectively, these data provide the first evidence that specific integrins regulate opioid receptor signaling in sensory neurons.
authors.affiliation: Department of Pharmacology, University of Texas Health Science Center, San Antonio, TX 78229-3900, USA.
authors.forename: K A
authors.initials: KA
authors.lastname: Berg
authors.forename: G
authors.initials: G
authors.lastname: Zardeneta
authors.forename: K M
authors.initials: KM
authors.lastname: Hargreaves
authors.forename: W P
authors.initials: WP
authors.lastname: Clarke
authors.forename: S B
authors.initials: SB
authors.lastname: Milam
mesh_terms: D000701:Analgesics, Opioid / Q000494:pharmacology; D000818:Animals; D000906:Antibodies / Q000494:pharmacology; D001920:Bradykinin / Q000378:metabolism / Q000494:pharmacology; D002478:Cells, Cultured; D000242:Cyclic AMP / Q000378:metabolism; D015232:Dinoprostone / Q000378:metabolism / Q000494:pharmacology; D020875:Enkephalin, Ala(2)-MePhe(4)-Gly(5)- / Q000494:pharmacology; D022001:Focal Adhesions / Q000378:metabolism; D019012:Integrin beta1 / Q000378:metabolism; D039661:Integrin beta3 / Q000378:metabolism; D016023:Integrins / Q000378:metabolism*; D008297:Male; D009475:Neurons, Afferent / Q000187:drug effects / Q000378:metabolism*; D009842:Oligopeptides / Q000494:pharmacology; D051381:Rats; D017207:Rats, Sprague-Dawley; D043562:Receptors, G-Protein-Coupled / Q000187:drug effects / Q000378:metabolism; D017450:Receptors, Opioid, mu / Q000378:metabolism*; D015398:Signal Transduction / Q000502:physiology*; D012668:Trigeminal Ganglion / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*
chemical_list: D000701:Analgesics, Opioid; D000906:Antibodies; D019012:Integrin beta1; D039661:Integrin beta3; D016023:Integrins; D009842:Oligopeptides; D043562:Receptors, G-Protein-Coupled; D017450:Receptors, Opioid, mu; D020875:Enkephalin, Ala(2)-MePhe(4)-Gly(5)-; C049554:glycyl-arginyl-glycyl-aspartyl-serine; D000242:Cyclic AMP; D015232:Dinoprostone; D001920:Bradykinin