The Journal of Physiological Sciences

, Volume 68, Issue 3, pp 261–268 | Cite as

The potentiating effect of calcitonin gene-related peptide on transient receptor potential vanilloid-1 activity and the electrophysiological responses of rat trigeminal neurons to nociceptive stimuli

  • Duangthip Chatchaisak
  • Mark Connor
  • Anan Srikiatkhachorn
  • Banthit Chetsawang
Original Paper


Growing evidence suggests that calcitonin gene-related peptide (CGRP) participates in trigeminal nociceptive responses. However, the role of CGRP in sensitization or desensitization of nociceptive transduction remains poorly understood. In this study, we sought to further investigate the CGRP-induced up-regulation of transient receptor potential vanilloid-1 (TRPV1) and the responses of trigeminal neurons to nociceptive stimuli. Rat trigeminal ganglion (TG) organ cultures and isolated trigeminal neurons were incubated with CGRP. An increase in TRPV1 levels was observed in CGRP-incubated TG organ cultures. CGRP potentiated capsaicin-induced increase in phosphorylated CaMKII levels in the TG organ cultures. The incubation of the trigeminal neurons with CGRP significantly increased the inward currents in response to capsaicin challenge, and this effect was inhibited by co-incubation with the CGRP receptor antagonist, BIBN4068BS or the inhibitor of protein kinase A, H-89. These findings reveal that CGRP acting on trigeminal neurons may play a significant role in facilitating cellular events that contribute to the peripheral sensitization of the TG in nociceptive transmission.


Trigeminal ganglion CGRP TRPV1 Capsaicin Nociceptive transmission 



This work was supported by grants from the Thailand Research Fund (TRF) to AS (Senior Scholar Fellowship), the TRF-Royal Golden Jubilee Ph.D. Program and the Office of the Higher Education Commission to DC and a TRF grant to the Institute of Molecular Biosciences (IRG5780009) and Mahidol University.

Author contributions

The individual contribution to the article of each author is declared following: (1) DC contributed to data acquisition and analysis and preparation of the drafting manuscript. (2) MC contributed to analysis and interpretation of data. (3) AS contributed to analysis and interpretation of data. (4) BC contributed to experimental design, analysis and interpretation of data and critically revised the manuscript.

Compliance with ethical standards

Conflict of interest

The authors have no financial or other relationship that could lead to a conflict of interest. None of the authors have any competing interests.

Ethical approval

All of the experiments in this study were conducted in accordance with the NIH Guidelines for the Care and Use of Animals, and the protocols were approved by the Institute of Molecular Biosciences Animal Care and Use Committee (MB-ACUC) of Mahidol University, Thailand (COA. No. MB-ACUC 2011/002) and the Macquarie University Animal Ethics Committee (approval 2012/058).


  1. 1.
    Anderson LE, Seybold VS (2004) Calcitonin gene-related peptide regulates gene transcription in primary afferent neurons. J Neurochem 91:1417–1429PubMedCrossRefGoogle Scholar
  2. 2.
    Fan PC, Kuo PH, Chang SH, Lee WT, Wu RM, Chiou LC (2009) Plasma calcitonin gene-related peptide in diagnosing and predicting pediatric migraine. Cephalalgia 29:883–890PubMedCrossRefGoogle Scholar
  3. 3.
    Juhasz G, Zsombok T, Modos EA, Olajos S, Jakab B, Nemeth J, Szolcsanyi J, Vitrai J, Bagdy G (2003) NO-induced migraine attack: strong increase in plasma calcitonin gene-related peptide (CGRP) concentration and negative correlation with platelet serotonin release. Pain 106:461–470PubMedCrossRefGoogle Scholar
  4. 4.
    Simonetti M, Giniatullin R, Fabbretti E (2008) Mechanisms mediating the enhanced gene transcription of P2X3 receptor by calcitonin gene-related peptide in trigeminal sensory neurons. J Biol Chem 283:18743–18752PubMedCrossRefGoogle Scholar
  5. 5.
    Chatchaisak D, Srikiatkhachorn A, le-Grand SM, Govitrapong P, Chetsawang B (2013) The role of calcitonin gene-related peptide on the increase in transient receptor potential vanilloid-1 levels in trigeminal ganglion and trigeminal nucleus caudalis activation of rat. J Chem Neuroanat 47:50–56PubMedCrossRefGoogle Scholar
  6. 6.
    Zhang Z, Winborn CS, de Prado BM, Russo AF (2007) Sensitization of calcitonin gene-related peptide receptors by receptor activity-modifying protein-1 in the trigeminal ganglion. J Neurosci 27:2693–2703PubMedCrossRefGoogle Scholar
  7. 7.
    Vause CV, Durham PL (2009) CGRP stimulation of iNOS and NO release from trigeminal ganglion glial cells involves mitogen-activated protein kinase pathways. J Neurochem 110:811–821PubMedPubMedCentralCrossRefGoogle Scholar
  8. 8.
    Capuano A, De Corato A, Lisi L, Tringali G, Navarra P, Dello Russo C (2009) Proinflammatory-activated trigeminal satellite cells promote neuronal sensitization: relevance for migraine pathology. Mol Pain 5:43PubMedPubMedCentralCrossRefGoogle Scholar
  9. 9.
    Cady RJ, Glenn JR, Smith KM, Durham PL (2011) Calcitonin gene-related peptide promotes cellular changes in trigeminal neurons and glia implicated in peripheral and central sensitization. Mol Pain 7:94PubMedPubMedCentralCrossRefGoogle Scholar
  10. 10.
    Bird GC, Han JS, Fu Y, Adwanikar H, Willis WD, Neugebauer V (2006) Pain-related synaptic plasticity in spinal dorsal horn neurons: role of CGRP. Mol Pain 2:31PubMedPubMedCentralCrossRefGoogle Scholar
  11. 11.
    Fischer MJ, Koulchitsky S, Messlinger K (2005) The nonpeptide calcitonin gene-related peptide receptor antagonist BIBN4096BS lowers the activity of neurons with meningeal input in the rat spinal trigeminal nucleus. J Neurosci 25:5877–5883PubMedCrossRefGoogle Scholar
  12. 12.
    Storer RJ, Akerman S, Goadsby PJ (2004) Calcitonin gene-related peptide (CGRP) modulates nociceptive trigeminovascular transmission in the cat. Br J Pharmacol 142:1171–1181PubMedPubMedCentralCrossRefGoogle Scholar
  13. 13.
    Chizh BA, O’Donnell MB, Napolitano A, Wang J, Brooke AC, Aylott MC, Bullman JN, Gray EJ, Lai RY, Williams PM, Appleby JM (2007) The effects of the TRPV1 antagonist SB-705498 on TRPV1 receptor-mediated activity and inflammatory hyperalgesia in humans. Pain 132:132–141PubMedCrossRefGoogle Scholar
  14. 14.
    Lee J, Saloman JL, Weiland G, Auh QS, Chung MK, Ro JY (2012) Functional interactions between NMDA receptors and TRPV1 in trigeminal sensory neurons mediate mechanical hyperalgesia in the rat masseter muscle. Pain 153:1514–1524PubMedPubMedCentralCrossRefGoogle Scholar
  15. 15.
    Durham PL, Vause CV (2010) Calcitonin gene-related peptide (CGRP) receptor antagonists in the treatment of migraine. CNS Drugs 24:539–548PubMedPubMedCentralCrossRefGoogle Scholar
  16. 16.
    Van Rossum D, Hanisch UK, Quirion R (1997) Neuroanatomical localization, pharmacological characterization and functions of CGRP, related peptides and their receptors. Neurosci Biobehav Rev 21:649–678PubMedCrossRefGoogle Scholar
  17. 17.
    Petersen KA, Nilsson E, Olesen J (2005) Presence and function of the calcitonin gene-related peptide receptor on rat pial arteries investigated in vitro and in vivo. Cephalalgia 25:424–432PubMedCrossRefGoogle Scholar
  18. 18.
    Gallai V, Sarchielli P, Floridi A, Franceschini M, Codini M, Glioti G, Trequattrini A, Palumbo R (1995) Vasoactive peptide levels in the plasma of young migraine patients with and without aura assessed both interictally and ictally. Cephalalgia 15:384–390PubMedCrossRefGoogle Scholar
  19. 19.
    Zygmunt PM, Petersson J, Andersson DA, Chuang H, Sørgård M, Di Marzo V, Julius D, Högestätt ED (1999) Vanilloid receptors on sensory nerves mediate the vasodilator action of anandamide. Nature 400:452–457PubMedCrossRefGoogle Scholar
  20. 20.
    Ji RR, Samad TA, Jin SX, Schmoll R, Woolf CJ (2002) p38 MAPK activation by NGF in primary sensory neurons after inflammation increases TRPV1 levels and maintains heat hyperalgesia. Neuron 26:57–68CrossRefGoogle Scholar
  21. 21.
    Lassen LH, Haderslev PA, Jacobsen VB, Iversen HK, Sperling B, Olesen J (2002) CGRP may play a causative role in migraine. Cephalalgia 22:54–61PubMedCrossRefGoogle Scholar
  22. 22.
    Tajti J, Kuris A, Vécsei L, Xu CB, Edvinsson L (2011) Organ culture of the trigeminal ganglion induces enhanced expression of calcitonin gene-related peptide via activation of extracellular signal-regulated protein kinase 1/2. Cephalalgia 31:95–105PubMedCrossRefGoogle Scholar
  23. 23.
    Petersen M, LaMotte RH (1993) Effect of protons on the inward current evoked by capsaicin in isolated dorsal root ganglion cells. Pain 54:37–42PubMedCrossRefGoogle Scholar
  24. 24.
    Fang L, Wu J, Lin Q, Willis WD (2002) Calcium-calmodulin-dependent protein kinase II contributes to spinal cord central sensitization. J Neurosci 22:4196–4204PubMedCrossRefGoogle Scholar
  25. 25.
    Ichikawa H, Gouty S, Regalia J, Helke CJ, Sugimoto T (2004) Ca2+/calmodulin-dependent protein kinase II in the rat cranial sensory ganglia. Brain Res 1005:36–43PubMedCrossRefGoogle Scholar
  26. 26.
    Meng J, Ovsepian SV, Wang J, Pickering M, Sasse A, Aoki KR, Lawrence GW, Dolly JO (2009) Activation of TRPV1 mediates calcitonin gene-related peptide release, which excites trigeminal sensory neurons and is attenuated by a retargeted botulinum toxin with anti-nociceptive potential. J Neurosci 29:4981–4992PubMedCrossRefGoogle Scholar
  27. 27.
    Price TJ, Jeske NA, Flores CM, Hargreaves KM (2005) Pharmacological interactions between calcium/calmodulin-dependent kinase II alpha and TRPV1 receptors in rat trigeminal sensory neurons. Neurosci Lett 389:94–98PubMedPubMedCentralCrossRefGoogle Scholar
  28. 28.
    Jung J, Shin JS, Lee SY, Hwang SW, Koo J, Cho H, Oh U (2004) Phosphorylation of vanilloid receptor 1 by Ca2+/calmodulin-dependent kinase II regulates its vanilloid binding. J Biol Chem 279:7048–7054PubMedCrossRefGoogle Scholar
  29. 29.
    Liang R, Liu X, Wei L, Wang W, Zheng P, Yan X, Zhao Y, Liu L, Cao X (2012) The modulation of the excitability of primary sensory neurons by Ca2+–CaM–CaMKII pathway. Neurol Sci 33:1083–1093PubMedCrossRefGoogle Scholar
  30. 30.
    Roberts LA, Christie MJ, Connor M (2002) Anandamide is a partial agonist at native vanilloid receptors in acutely isolated mouse trigeminal sensory neurons. Br J Pharmacol 137:421–428PubMedPubMedCentralCrossRefGoogle Scholar
  31. 31.
    Ichikawa T, Ishihara K, Kusakabe T, Hiruma H, Kawakami T, Hotta K (2000) CGRP modulates mucin synthesis in surface mucus cells of rat gastric oxyntic mucosa. Am J Physiol Gastrointest Liver Physiol 279:G82–G89PubMedCrossRefGoogle Scholar
  32. 32.
    Devesa I, Ferrándiz-Huertas C, Mathivanan S, Wolf C, Luján R, Changeux JP, Ferrer-Montiel A (2014) αCGRP is essential for algesic exocytotic mobilization of TRPV1 channels in peptidergic nociceptors. Proc Natl Acad Sci USA 111:18345–18350PubMedPubMedCentralCrossRefGoogle Scholar
  33. 33.
    Lennerz J, Ruhle V, Ceppa EP, Neuhuber WL, Bunnett NW, Grady EF, Messlinger K (2008) Calcitonin receptor-like receptor (CLR), receptor activity-modifying protein 1 (RAMP1), and calcitonin gene-related peptide (CGRP) immunoreactivity in the rat trigeminovascular system: differences between peripheral and central CGRP receptor distribution. J Comp Neurol 507:1277–1299PubMedCrossRefGoogle Scholar
  34. 34.
    Sun RQ, Tun YJ, Lawand NB, Yan JY, Lin Q, Willis WD (2004) Calcitonin gene-related peptide receptor activation produces PKA- and PKC-dependent mechanical hyperalgesia and central sensitization. J Neurophysiol 92:2859–2866PubMedCrossRefGoogle Scholar
  35. 35.
    Pan HL, Zhang YQ, Zhao ZQ (2010) Involvement of lysophosphatidic acid in bone cancer pain by potentiation of TRPV1 via PKCε pathway in dorsal root ganglion neurons. Mol Pain 92:2859–2866Google Scholar
  36. 36.
    Seybold VS, McCarson KE, Mermelstein PG, Groth RD, Abrahams LG (2003) Calcitonin gene-related peptide regulates expression of neurokinin 1 receptors by rat spinal neurons. J Neurosci 23:1816–1824PubMedCrossRefGoogle Scholar

Copyright information

© The Physiological Society of Japan and Springer Japan 2017

Authors and Affiliations

  • Duangthip Chatchaisak
    • 1
  • Mark Connor
    • 2
  • Anan Srikiatkhachorn
    • 3
  • Banthit Chetsawang
    • 1
  1. 1.Research Center for Neuroscience, Institute of Molecular BiosciencesMahidol UniversitySalayaThailand
  2. 2.Australian School of Advanced MedicineMacquarie UniversitySydneyAustralia
  3. 3.International Medical CollegeKing Mongkut’s Institute of Technology LadkrabangBangkokThailand

Personalised recommendations