title: Gα(q/11) signaling tonically modulates nociceptor function and contributes to activity-dependent sensitization.
abstract: Peripheral injury or inflammation leads to a release of mediators capable of binding to a variety of ion channels and receptors. Among these are the 7-transmembrane receptors (G protein-coupled receptors) coupling to G(s), G(i/o), G₁₂/₁₃, or G(q/11) G proteins. Each of the G protein-coupled receptor pathways is involved in nociceptive modulation and pain processing, but the relative contribution of individual signaling pathways in vivo has not yet been worked out. The G(q)/G₁₁ signaling branch is of particular interest because it leads to the activation of phospholipase C-β, protein kinase C, the release of calcium from intracellular stores, and it modulates extracellular regulated kinases. To investigate the contribution of the entire G(q/11)-signaling pathway in nociceptors towards regulation of pain, we generated double-deficient mice lacking G(q/11) selectively in nociceptors using a conditional gene-targeting approach. We observed that nociceptor-specific loss of G(q) and G₁₁ results in reduced pain hypersensitivity following paw inflammation or spared nerve injury. Surprisingly, our behavioral and electrophysiological experiments also indicated defects in basal mechanical sensitivity in G(q/11) mutant mice, suggesting a novel function for G(q/11) in tonic modulation of acute nociception. Patch-clamp recordings revealed changes in voltage-dependent tetrodotoxin-resistant and tetrodotoxin-sensitive sodium channels in nociceptors upon a loss of G(q/11), whereas potassium currents remained unchanged. Our results indicate that the functional role of the G(q)/G₁₁ branch of G-protein signaling in nociceptors in vivo not only spans sensitization mechanisms in pathological pain states, but is also operational in tonic modulation of basal nociception and acute pain.
authors.affiliation: Pharmacology Institut, University of Heidelberg, Heidelberg, Germany Department of Physiology and Medical Physics, Division of Physiology, Innsbruck Medical University, Innsbruck, Austria Pharmazentrum Frankfurt, Klinikum der Goethe-Universität, Frankfurt am Main 60590, Germany Max-Delbrueck-Center for Molecular Medicine, Department of Neurosciences, Berlin 13125, Germany Spinal Cord Group, West Medical Building, University of Glasgow, University Avenue, Glasgow, UK.
authors.forename: Anke
authors.initials: A
authors.lastname: Tappe-Theodor
authors.forename: Cristina E
authors.initials: CE
authors.lastname: Constantin
authors.forename: Irmgard
authors.initials: I
authors.lastname: Tegeder
authors.forename: Stefan G
authors.initials: SG
authors.lastname: Lechner
authors.forename: Michiel
authors.initials: M
authors.lastname: Langeslag
authors.forename: Peter
authors.initials: P
authors.lastname: Lepcynzsky
authors.forename: Richard I
authors.initials: RI
authors.lastname: Wirotanseng
authors.forename: Martina
authors.initials: M
authors.lastname: Kurejova
authors.forename: Nitin
authors.initials: N
authors.lastname: Agarwal
authors.forename: Gergely
authors.initials: G
authors.lastname: Nagy
authors.forename: Andrew
authors.initials: A
authors.lastname: Todd
authors.forename: Nina
authors.initials: N
authors.lastname: Wettschureck
authors.forename: Stefan
authors.initials: S
authors.lastname: Offermanns
authors.forename: Michaela
authors.initials: M
authors.lastname: Kress
authors.forename: Gary R
authors.initials: GR
authors.lastname: Lewin
authors.forename: Rohini
authors.initials: R
authors.lastname: Kuner
mesh_terms: D000818:Animals; D002478:Cells, Cultured; D043802:GTP-Binding Protein alpha Subunits, Gq-G11 / Q000235:genetics / Q000378:metabolism*; D005727:Ganglia, Spinal / Q000166:cytology / Q000378:metabolism* / Q000503:physiopathology; D006930:Hyperalgesia / Q000378:metabolism* / Q000503:physiopathology; D051379:Mice; D018345:Mice, Knockout; D009474:Neurons / Q000166:cytology / Q000502:physiology*; D059225:Nociception / Q000502:physiology*; D009619:Nociceptors / Q000502:physiology*; D010147:Pain Measurement; D011493:Protein Kinase C / Q000378:metabolism; D015398:Signal Transduction / Q000502:physiology; D010738:Type C Phospholipases / Q000378:metabolism
chemical_list: D011493:Protein Kinase C; D010738:Type C Phospholipases; D043802:GTP-Binding Protein alpha Subunits, Gq-G11
references.citation: Abrahamsen B, Zhao J, Asante CO, Cendan CM, Marsh S, Martinez-Barbera JP, Nassar MA, Dickenson AH, Wood JN. The cell and molecular basis of mechanical, cold, and inflammatory pain. Science. 2008;321:702-705.
references.citation: Agarwal N, Offermanns S, Kuner R. Conditional gene deletion in primary nociceptive neurons of trigeminal ganglia and dorsal root ganglia. Genesis. 2004;38:122-129.
references.citation: Agarwal N, Pacher P, Tegeder I, Amaya F, Constantin CE, Brenner GJ, Rubino T, Michalski CW, Marsicano G, Monory K, Mackie K, Marian C, Batkai S, Parolaro D, Fischer MJ, Reeh P, Kunos G, Kress M, Lutz B, Woolf CJ, Kuner R. Cannabinoids mediate analgesia largely via peripheral type 1 cannabinoid receptors in nociceptors. Nat Neurosci. 2007;10:870-879.
references.citation: Akopian AN, Souslova V, England S, Okuse K, Ogata N, Ure J, Smith A, Kerr BJ, McMahon SB, Boyce S, Hill R, Stanfa LC, Dickenson AH, Wood JN. The tetrodotoxin-resistant sodium channel SNS has a specialized function in pain pathways. Nat Neurosci. 1999;2:541-548.
references.citation: Bhave G, Hu HJ, Glauner KS, Zhu W, Wang H, Brasier DJ, Oxford GS, Gereau RW 4th. Protein kinase C phosphorylation sensitizes but does not activate the capsaicin receptor transient receptor potential vanilloid 1 (TRPV1). Proc Natl Acad Sci USA. 2003;100:12480-12485.
references.citation: Bockaert J, Perroy J, Becamel C, Marin P, Fagni L. GPCR interacting proteins (GIPs) in the nervous system: roles in physiology and pathologies. Annu Rev Pharmacol Toxicol. 2002;50:89-109.
references.citation: Brown DA, Passmore GM. Neural KCNQ (Kv7) channels. Br J Pharmacol. 2009;156:1185-1195.
references.citation: Cavanaugh DJ, Lee H, Lo L, Shields SD, Zylka MJ, Basbaum AI, Anderson DJ. Distinct subsets of unmyelinated primary sensory fibers mediate behavioral responses to noxious thermal and mechanical stimuli. Proc Natl Acad Sci USA. 2009;106:9075-9080.
references.citation: Cesare P, Dekker LV, Sardini A, Parker PJ, McNaughton PA. Specific involvement of PKC-epsilon in sensitization of the neuronal response to painful heat. Neuron. 1999;23:617-624.
references.citation: Chen C, Clarke IJ. G(o)-2 protein mediates the reduction in Ca2+ currents by somatostatin in cultured ovine somatotrophs. J Physiol. 1996;491:21-29.
references.citation: Chen X, Talley EM, Patel N, Gomis A, McIntire WE, Dong B, Viana F, Garrison JC, Bayliss DA. Inhibition of a background potassium channel by Gq protein alpha-subunits. Proc Natl Acad Sci USA. 2006;103:3422-3427.
references.citation: Chuang HH, Prescott ED, Kong H, Shields S, Jordt SE, Basbaum AI, Chao MV, Julius D. Bradykinin and nerve growth factor release the capsaicin receptor from PtdIns(4,5)P2-mediated inhibition. Nature. 2001;411:957-962.
references.citation: Constantin CE, Mair N, Sailer CA, Andratsch M, Xu ZZ, Blumer MJ, Scherbakov N, Davis JB, Bluethmann H, Ji RR, Kress M. Endogenous tumor necrosis factor alpha (TNFalpha) requires TNF receptor type 2 to generate heat hyperalgesia in a mouse cancer model. J Neurosci. 2008;28:5072-5081.
references.citation: Decosterd I, Woolf CJ. Spared nerve injury: an animal model of persistent peripheral neuropathic pain. Pain. 2000;87:149-158.
references.citation: Dittert I, Vlachová V, Knotková H, Vitásková Z, Vyklicky L, Kress M, Reeh PW. A technique for fast application of heated solutions of different composition to cultured neurones. J Neurosci Methods. 2008;82:195-201.
references.citation: Drew L, Wood J, Cesare P. Distinct mechanosensitive properties of capsaicin-sensitive and -insensitive sensory neurons. J Neurosci. 2002;22:RC228.
references.citation: Drissi H, Lasmoles F, Le Mellay V, Marie PJ, Lieberherr M. Activation of phospholipase C-beta1 via Galphaq/11 during calcium mobilization by calcitonin gene-related peptide. J Biol Chem. 1998;273:20168-20174.
references.citation: Forster C, Handwerker HO. Automatic classification and analysis of microneurographic spike data using a PC/AT. J Neurosci Methods. 1990;31:109-118.
references.citation: Gamper N, Reznikov V, Yamada Y, Yang J, Shapiro MS. Phosphatidylinositol [correction] 4,5-bisphosphate signals underlie receptor-specific Gq/11-mediated modulation of N-type Ca2+ channels. J Neurosci. 2004;24:10980-10992.
references.citation: Ghahremani MH, Forget C, Albert PR. Distinct roles for Gαi2 and Gβγ in signaling to DNA synthesis and Gαi3 in cellular transformation by dopamine D2S receptor activation in BALB/c 3T3 cells. Mol Cell Biol. 2000;20:1497-1506.
references.citation: Gold MS. Tetrodotoxin-resistant Na+ currents and inflammatory hyperalgesia. Proc Natl Acad Sci USA. 1999;96:7645-7649.
references.citation: Gold MS, Reichling DB, Shuster MJ, Levine JD. Hyperalgesic agents increase a tetrodotoxin-resistant Na+ current in nociceptors. Proc Natl Acad Sci USA. 1996;93:1108-1112.
references.citation: Goudet C, Magnaghi V, Landry M, Nagy F, Gereau RW 4th, Pin JP. Metabotropic receptors for glutamate and GABA in pain. Brain Res Rev. 2009;60:43-56.
references.citation: Han SK, Mancino V, Simon MI. Phospholipase Cbeta 3 mediates the scratching response activated by the histamine H1 receptor on C-fiber nociceptive neurons. Neuron. 2006;52:691-703.
references.citation: Hartmann J, Blum R, Kovalchuk Y, Adelsberger H, Kuner R, Durand GM, Miyata M, Kano M, Offermanns S, Konnerth A. Distinct roles of Galpha(q) and Galpha11 for Purkinje cell signaling and motor behavior. J Neurosci. 2004;24:5119-5130.
references.citation: Hartmann B, Ahmadi S, Heppenstall PA, Lewin GR, Schott C, Borchardt T, Seeburg PH, Zeilhofer HU, Sprengel R, Kuner R. The AMPA receptor subunits GluR-A and GluR-B reciprocally modulate spinal synaptic plasticity and inflammatory pain. Neuron. 2004;44:637-650.
references.citation: Honoré E. The neuronal background K2P channels: focus on TREK1. Nat Rev Neurosci. 2007;8:251-261.
references.citation: Hu J, Lewin G. Mechanosensitive currents in the neurites of cultured mouse sensory neurones. J Physiol. 2006;577:815-828.
references.citation: Huang J, Zhang X, McNaughton PA. Inflammatory pain: the cellular basis of heat hyperalgesia. Curr Neuropharmacol. 2006;4:197-206.
references.citation: Hucho T, Levine JD. Signaling pathways in sensitization: toward a nociceptor cell biology. Neuron. 2007;55:365-376.
references.citation: Jiang M, Spicher K, Boulay G, Wang Y, Birnbaumer L. Most central nervous system D2 dopamine receptors are coupled to their effectors by Go. Proc Natl Acad Sci USA. 2001;98:3577-3582.
references.citation: Kenakin T. G protein coupled receptors as allosteric proteins and the role of allosteric modulators. J Recept Signal Transduct Res. 2010;30:313-321.
references.citation: Kleppisch T, Voigt V, Allmann R, Offermanns S. G(alpha)q-deficient mice lack metabotropic glutamate receptor-dependent long-term depression but show normal long-term potentiation in the hippocampal CA1 region. J Neurosci. 2001;21:4943-4948.
references.citation: Kress M, Koltzenburg M, Reeh PW, Handwerker HO. Responsiveness and functional attributes of electrically localized terminals of cutaneous C-fibers in vivo and in vitro. J Neurophysiol. 1992;68:581-595.
references.citation: Kwan KY, Allchorne AJ, Vollrath MA, Christensen AP, Zhang DS, Woolf CJ, Corey DP. TRPA1 contributes to cold, mechanical, and chemical nociception but is not essential for hair-cell transduction. Neuron. 2006;50:277-289.
references.citation: Macpherson LJ, Xiao B, Kwan KY, Petrus MJ, Dubin AE, Hwang S, Cravatt B, Corey DP, Patapoutian A. An ion channel essential for sensing chemical damage. J Neurosci. 2007;27:11412-11415.
references.citation: Mathie A. Neuronal two-pore-domain potassium channels and their regulation by G protein-coupled receptors. J Physiol. 2007;578:377-385.
references.citation: McNamara CR, Mandel-Brehm J, Bautista DM, Siemens J, Deranian KL, Zhao M, Hayward NJ, Chong JA, Julius D, Moran MM, Fanger CM. TRPA1 mediates formalin-induced pain. Proc Natl Acad Sci USA. 2007;104:13525-13530.
references.citation: Moers A, Nieswandt B, Massberg S, Wettschureck N, Gruner S, Konrad I, Schulte V, Aktas B, Gratacap MP, Simon MI, Gawaz M, Offermanns S. G13 is an essential mediator of platelet activation in hemostasis and thrombosis. Nat Med. 2003;9:1418-1422.
references.citation: Nagata K, Ye C, Jain M, Milstone DS, Liao R, Mortensen RM. Galpha(i2) but not Galpha(i3) is required for muscarinic inhibition of contractility and calcium currents in adult cardiomyocytes. Circ Res. 2000;87:903-909.
references.citation: Nicol GD, Vasko MR. Unraveling the story of NGF-mediated sensitization of nociceptive sensory neurons: ON or OFF the Trks? Mol Interv. 2007;7:26-41.
references.citation: Obreja O, Rathee PK, Lips KS, Distler C, Kress M. IL-1 beta potentiates heat-activated currents in rat sensory neurons: involvement of IL-1RI, tyrosine kinase, and protein kinase C. FASEB J. 2002;16:1497-1503.
references.citation: Obreja O, Biasio W, Andratsch M, Lips KS, Rathee PK, Ludwig A, Rose-John S, Kress M. Fast modulation of heat-activated ionic current by proinflammatory interleukin 6 in rat sensory neurons. Brain. 2005;128:1634-1641.
references.citation: Offermanns S, Toombs CF, Hu YH, Simon MI. Defective platelet activation in G alpha(q)-deficient mice. Nature. 1997;389:183-186.
references.citation: Offermanns S, Zhao LP, Gohla A, Sarosi I, Simon MI, Wilkie TM. Embryonic cardiomyocyte hypoplasia and craniofacial defects in G alpha q/G alpha 11-mutant mice. EMBO J. 1998;17:4304-4312.
references.citation: Reichling DB, Levine JD. Critical role of nociceptor plasticity in chronic pain. Trends Neurosci. 2009;32:611-618.
references.citation: Ronnett GV, Moon C. G proteins and olfactory signal transduction. Annu Rev Physiol. 2002;64:189-222.
references.citation: Rosenbaum DM, Rasmussen SG, Kobilka BK. The structure and function of G-protein-coupled receptors. Nature. 2009;459:356-363.
references.citation: Schmidt M, Dubin AE, Petrus MJ, Earley TJ, Patapoutian A. Nociceptive signals induce trafficking of TRPA1 to the plasma membrane. Neuron. 2009;64:498-509.
references.citation: Shields SD, Cavanaugh DJ, Lee H, Anderson DJ, Basbaum AI. Pain behavior in the formalin test persists after ablation of the great majority of C-fiber nociceptors. Pain. 2010;151:422-429.
references.citation: Sun Y, Huang J, Xiang Y, Bastepe M, Jüppner H, Kobilka BK, Zhang JJ, Huang XY. Dosage-dependent switch from G protein-coupled to G protein-independent signaling by a GPCR. EMBO J. 2007;26:53-64.
references.citation: Stösser S, Agarwal N, Tappe-Theodor A, Yanagisawa M, Kuner R. Dissecting the functional significance of endothelin A receptors in peripheral nociceptors in vivo via conditional gene deletion. Pain. 2010;148:206-214.
references.citation: Wettschureck N, Rütten H, Zywietz A, Gehring D, Wilkie TM, Chen J, Chien KR, Offermanns S. Absence of pressure overload induced myocardial hypertrophy after conditional inactivation of Galphaq/Galpha11 in cardiomyocytes. Nat Med. 2001;7:1236-1240.
references.citation: Wettschureck N, Offermanns S. Mammalian G proteins and their cell type specific functions. Physiol Rev. 2005;85:1159-1204.
references.citation: Wettschureck N, Moers A, Wallenwein B, Parlow AF, Maser-Gluth C, Offermanns S. Loss of Gq/11 family G proteins in the nervous system causes pituitary somatotroph hypoplasia and dwarfism in mice. Mol Cell Biol. 2005;25:1942-1948.
references.citation: Wirth A, Benyó Z, Lukasova M, Leutgeb B, Wettschureck N, Gorbey S, Orsy P, Horváth B, Maser-Gluth C, Greiner E, Lemmer B, Schütz G, Gutkind JS, Offermanns S. G12-G13-LARG-mediated signaling in vascular smooth muscle is required for salt-induced hypertension. Nat Med. 2008;14:64-68.
references.citation: Wood JN, Boorman JP, Okuse K, Baker MD. Voltage-gated sodium channels and pain pathways. J Neurobiol. 2004;61:55-71.
references.citation: Yasuda N, Miura S, Akazawa H, Tanaka T, Qin Y, Kiya Y, Imaizumi S, Fujino M, Ito K, Zou Y, Fukuhara S, Kunimoto S, Fukuzaki K, Sato T, Ge J, Mochizuki N, Nakaya H, Saku K, Komuro I. Conformational switch of angiotensin II type 1 receptor underlying mechanical stress-induced activation. EMBO Rep. 2008;9:179-186.
references.citation: Zhang X, Li L, McNaughton PA. Proinflammatory mediators modulate the heat-activated ion channel TRPV1 via the scaffolding protein AKAP79/150. Neuron. 2008;59:450-461.
references.citation: Zurborg S, Yurgionas B, Jira JA, Caspani O, Heppenstall PA. Direct activation of the ion channel TRPA1 by Ca2+. Nat Neurosci. 2007;10:277-279.