The Cerebellum

, Volume 12, Issue 6, pp 937–949 | Cite as

Localization of CGRP Receptor Components, CGRP, and Receptor Binding Sites in Human and Rhesus Cerebellar Cortex

  • Sajedeh Eftekhari
  • Christopher A. Salvatore
  • Renee C. Gaspar
  • Rhonda Roberts
  • Stacey O’Malley
  • Zhizhen Zeng
  • Lars Edvinsson
Original Paper


The cerebellum is classically considered to be mainly involved in motor processing, but studies have suggested several other functions, including pain processing. Calcitonin-gene-related peptide (CGRP) is a neuropeptide involved in migraine pathology, where there is elevated release of CGRP during migraine attacks and CGRP receptor antagonists have antimigraine efficacy. In the present study, we examined CGRP and CGRP receptor binding sites and protein expression in primate cerebellar cortex. Additionally, mRNA expression of the CGRP receptor components, calcitonin receptor-like receptor (CLR) and receptor activity modifying protein 1 (RAMP1), was examined. In addition, expression of procalcitonin was studied. We observed high [3H]MK-3207 (CGRP receptor antagonist) binding densities in the molecular layer of rhesus cerebellar cortex; however, due to the limit of resolution of the autoradiographic image the exact cellular localization could not be determined. Similarly, [125I]CGRP binding was observed in the molecular layer and Purkinje cell layer of human cerebellum. CLR and RAMP1 mRNA was expressed within the Purkinje cell layer and some expression was found in the molecular layer. Immunofluorescence revealed expression of CGRP, CLR, and RAMP1 in the Purkinje cells and in cells in the molecular layer. Procalcitonin was found in the same localization. Recent research in the biology of cerebellum indicates that it may have a role in nociception. For the first time we have identified CGRP and CGRP receptor binding sites together with CGRP receptor expression through protein and mRNA localization in primate cerebellar cortex. These results point toward a functional role of CGRP in cerebellum. Further efforts are needed to evaluate this.


Cerebellum Nociception CGRP CLR RAMP1 Primate 



Thanks are due to the following from Merck Research Laboratories: Stephanie Villarreal and John Majercak for their consultation on design and methodology details. Kenneth Lodge for collecting the rhesus tissue, Brett Connolly for H&E staining, and Patricia Miller for tissue preparation. Dr. Elisabet Englund at Lund University, Sweden, and Dr. Janos Tajiti at Albert Szent-Györgyi University Medical School, Szeged, Hungary, for providing the human tissue.

Conflict of Interest

The authors do not have any conflicts of interest related to this work.


  1. 1.
    Moulton EA, Schmahmann JD, Becerra L, Borsook D. The cerebellum and pain: passive integrator or active participator? Brain Res Rev. 2010;65:14–27.PubMedCrossRefGoogle Scholar
  2. 2.
    Teune TM, van der Burg J, van der Moer J, Voogd J, Ruigrok TJ. Topography of cerebellar nuclear projections to the brain stem in the rat. Prog Brain Res. 2000;124:141–72.PubMedCrossRefGoogle Scholar
  3. 3.
    Strick PL, Dum RP, Fiez JA. Cerebellum and nonmotor function. Annu Rev Neurosci. 2009;32:413–34.PubMedCrossRefGoogle Scholar
  4. 4.
    Apkarian AV, Bushnell MC, Treede RD, Zubieta JK. Human brain mechanisms of pain perception and regulation in health and disease. Eur J Pain. 2005;9:463–84.PubMedCrossRefGoogle Scholar
  5. 5.
    Schmahmann JD. From movement to thought: anatomic substrates of the cerebellar contribution to cognitive processing. Hum Brain Mapp. 1996;4:174–98.PubMedCrossRefGoogle Scholar
  6. 6.
    Edgley SA, Gallimore CM. The morphology and projections of dorsal horn spinocerebellar tract neurones in the cat. J Physiol. 1988;397:99–111.PubMedGoogle Scholar
  7. 7.
    Ekerot CF, Oscarsson O, Schouenborg J. Stimulation of cat cutaneous nociceptive C fibres causing tonic and synchronous activity in climbing fibres. J Physiol. 1987;386:539–46.PubMedGoogle Scholar
  8. 8.
    Ekerot CF, Gustavsson P, Oscarsson O, Schouenborg J. Climbing fibres projecting to cat cerebellar anterior lobe activated by cutaneous A and C fibres. J Physiol. 1987;386:529–38.PubMedGoogle Scholar
  9. 9.
    House E, Pansky B. A functional approach to neuroanatomy. New York: McGraw-Hill Book Company, Inc, New York; 1967Google Scholar
  10. 10.
    Jie W, Pei-Xi C. Discharge response of cerebellar Purkinje cells to stimulation of C-fiber in cat saphenous nerve. Brain Res. 1992;581:269–72.PubMedCrossRefGoogle Scholar
  11. 11.
    Fields HL. Pain modulation: expectation, opioid analgesia and virtual pain. Prog Brain Res. 2000;122:245–53.PubMedCrossRefGoogle Scholar
  12. 12.
    Weiller C, May A, Limmroth V, Juptner M, Kaube H, Schayck RV, et al. Brain stem activation in spontaneous human migraine attacks. Nat Med. 1995;1:658–60.PubMedCrossRefGoogle Scholar
  13. 13.
    Bahra A, Matharu MS, Buchel C, Frackowiak RS, Goadsby PJ. Brainstem activation specific to migraine headache. Lancet. 2001;357:1016–7.PubMedCrossRefGoogle Scholar
  14. 14.
    Borsook D, Moulton EA, Tully S, Schmahmann JD, Becerra L. Human cerebellar responses to brush and heat stimuli in healthy and neuropathic pain subjects. Cerebellum. 2008;7:252–72.PubMedCrossRefGoogle Scholar
  15. 15.
    Pietrobon D. Migraine: new molecular mechanisms. Neuroscientist. 2005;11:373–86.PubMedCrossRefGoogle Scholar
  16. 16.
    Skofitsch G, Jacobowitz DM. Calcitonin gene-related peptide: detailed immunohistochemical distribution in the central nervous system. Peptides. 1985;6:721–45.PubMedCrossRefGoogle Scholar
  17. 17.
    Hokfelt T, Arvidsson U, Ceccatelli S, Cortes R, Cullheim S, Dagerlind A, et al. Calcitonin gene-related peptide in the brain, spinal cord, and some peripheral systems. Ann N Y Acad Sci. 1992;657:119–34.PubMedCrossRefGoogle Scholar
  18. 18.
    Bellamy JL, Cady RK, Durham PL. Salivary levels of CGRP and VIP in rhinosinusitis and migraine patients. Headache. 2006;46:24–33.PubMedCrossRefGoogle Scholar
  19. 19.
    Goadsby PJ, Edvinsson L. The trigeminovascular system and migraine: studies characterizing cerebrovascular and neuropeptide changes seen in humans and cats. Ann Neurol. 1993;33:48–56.PubMedCrossRefGoogle Scholar
  20. 20.
    McLatchie LM, Fraser NJ, Main MJ, Wise A, Brown J, Thompson N, et al. RAMPs regulate the transport and ligand specificity of the calcitonin-receptor-like receptor. Nature. 1998;393:333–9.PubMedCrossRefGoogle Scholar
  21. 21.
    Evans BN, Rosenblatt MI, Mnayer LO, Oliver KR, Dickerson IM. CGRP-RCP, a novel protein required for signal transduction at calcitonin gene-related peptide and adrenomedullin receptors. J Biol Chem. 2000;275:31438–43.PubMedCrossRefGoogle Scholar
  22. 22.
    Zhang Z, Winborn CS, Marquez de Prado B, Russo AF. Sensitization of calcitonin gene-related peptide receptors by receptor activity-modifying protein-1 in the trigeminal ganglion. J Neurosci. 2007;27:2693–703.PubMedCrossRefGoogle Scholar
  23. 23.
    Ho TW, Edvinsson L, Goadsby PJ. CGRP and its receptors provide new insights into migraine pathophysiology. Nat Rev Neurol. 2010;6:573–82.PubMedCrossRefGoogle Scholar
  24. 24.
    Edvinsson L, Eftekhari S, Salvatore CA, Warfvinge K. Cerebellar distribution of calcitonin gene-related peptide (CGRP) and its receptor components calcitonin receptor-like receptor (CLR) and receptor activity modifying protein 1 (RAMP1) in rat. Mol Cell Neurosci. 2010;46:333–9.PubMedCrossRefGoogle Scholar
  25. 25.
    Hewitt DJ, Aurora SK, Dodick DW, Goadsby PJ, Ge YJ, Bachman R, et al. Randomized controlled trial of the CGRP receptor antagonist MK-3207 in the acute treatment of migraine. Cephalalgia. 2011;31:712–22.PubMedCrossRefGoogle Scholar
  26. 26.
    Salvatore CA, Moore EL, Calamari A, Cook JJ, Michener MS, O'Malley S, et al. Pharmacological properties of MK-3207, a potent and orally active calcitonin gene-related peptide receptor antagonist. J Pharmacol Exp Ther. 2010;333:152–60.PubMedCrossRefGoogle Scholar
  27. 27.
    Paone DV, Nguyen DN, Shaw AW, Burgey CS, Potteiger CM, Deng JZ, et al. Orally bioavailable imidazoazepanes as calcitonin gene-related peptide (CGRP) receptor antagonists: discovery of MK-2918. Bioorg Med Chem Lett. 2011;21:2683–6.PubMedCrossRefGoogle Scholar
  28. 28.
    Hershey JC, Corcoran HA, Baskin EP, Salvatore CA, Mosser S, Williams TM, et al. Investigation of the species selectivity of a nonpeptide CGRP receptor antagonist using a novel pharmacodynamic assay. Regul Pept. 2005;127:71–7.Google Scholar
  29. 29.
    Eftekhari S, Salvatore CA, Calamari A, Kane SA, Tajti J, Edvinsson L. Differential distribution of calcitonin gene-related peptide and its receptor components in the human trigeminal ganglion. Neuroscience. 2010;169:683–96.PubMedCrossRefGoogle Scholar
  30. 30.
    Vincent M, Hadjikhani N. The cerebellum and migraine. Headache. 2007;47:820–33.PubMedCrossRefGoogle Scholar
  31. 31.
    Saab CY, Willis WD. The cerebellum: organization, functions and its role in nociception. Brain Res Brain Res Rev. 2003;42:85–95.PubMedCrossRefGoogle Scholar
  32. 32.
    Edvinsson L, Linde M. New drugs in migraine treatment and prophylaxis: telcagepant and topiramate. Lancet. 2010;376:645–55.PubMedCrossRefGoogle Scholar
  33. 33.
    Helmchen C, Mohr C, Erdmann C, Petersen D, Nitschke MF. Differential cerebellar activation related to perceived pain intensity during noxious thermal stimulation in humans: a functional magnetic resonance imaging study. Neurosci Lett. 2003;335:202–6.PubMedCrossRefGoogle Scholar
  34. 34.
    Helmchen C, Mohr C, Erdmann C, Binkofski F. Cerebellar neural responses related to actively and passively applied noxious thermal stimulation in human subjects: a parametric fMRI study. Neurosci Lett. 2004;361:237–40.PubMedCrossRefGoogle Scholar
  35. 35.
    F.H. Maniyar TS, C. Schankin, T. Monteith, P.J. Goadsby. Investigating the premonitory phase of migraine with H2 15O PET. Cephalalgia 2013;33 SupplementGoogle Scholar
  36. 36.
    Sandor PS, Mascia A, Seidel L, de Pasqua V, Schoenen J. Subclinical cerebellar impairment in the common types of migraine: a three-dimensional analysis of reaching movements. Ann Neurol. 2001;49:668–72.PubMedCrossRefGoogle Scholar
  37. 37.
    Olesen J, Diener HC, Husstedt IW, Goadsby PJ, Hall D, Meier U, et al. Calcitonin gene-related peptide receptor antagonist BIBN 4096 BS for the acute treatment of migraine. N Engl J Med. 2004;350:1104–10.PubMedCrossRefGoogle Scholar
  38. 38.
    Ho TW, Ferrari MD, Dodick DW, Galet V, Kost J, Fan X, et al. Efficacy and tolerability of MK-0974 (telcagepant), a new oral antagonist of calcitonin gene-related peptide receptor, compared with zolmitriptan for acute migraine: a randomised, placebo-controlled, parallel-treatment trial. Lancet. 2008;372:2115–23.PubMedCrossRefGoogle Scholar
  39. 39.
    Kawai Y, Takami K, Shiosaka S, Emson PC, Hillyard CJ, Girgis S, et al. Topographic localization of calcitonin gene-related peptide in the rat brain: an immunohistochemical analysis. Neuroscience. 1985;15:747–63.PubMedCrossRefGoogle Scholar
  40. 40.
    Gregg KV, Bishop GA, King JS. Fine structural analysis of calcitonin gene-related peptide in the mouse inferior olivary complex. J Neurocytol. 1999;28:431–8.PubMedCrossRefGoogle Scholar
  41. 41.
    van Rossum D, Hanisch UK, Quirion R. Neuroanatomical localization, pharmacological characterization and functions of CGRP, related peptides and their receptors. Neurosci Biobehav Rev. 1997;21:649–78.PubMedCrossRefGoogle Scholar
  42. 42.
    Morara S, Wimalawansa SJ, Rosina A. Monoclonal antibodies reveal expression of the CGRP receptor in Purkinje cells, interneurons and astrocytes of rat cerebellar cortex. Neuroreport. 1998;9:3755–9.PubMedCrossRefGoogle Scholar
  43. 43.
    Briatore F, Patrizi A, Viltono L, Sassoe-Pognetto M, Wulff P. Quantitative organization of GABAergic synapses in the molecular layer of the mouse cerebellar cortex. PLoS One. 2010;5:e12119.PubMedCrossRefGoogle Scholar
  44. 44.
    van Rossum D, Menard DP, Fournier A, St-Pierre S, Quirion R. Autoradiographic distribution and receptor binding profile of [125I]Bolton Hunter-rat amylin binding sites in the rat brain. J Pharmacol Exp Ther. 1994;270:779–87.PubMedGoogle Scholar
  45. 45.
    Sexton PM, Paxinos G, Kenney MA, Wookey PJ, Beaumont K. In vitro autoradiographic localization of amylin binding sites in rat brain. Neuroscience. 1994;62:553–67.PubMedCrossRefGoogle Scholar
  46. 46.
    Morara S, Rosina A, Provini L, Forloni G, Caretti A, Wimalawansa SJ. Calcitonin gene-related peptide receptor expression in the neurons and glia of developing rat cerebellum: an autoradiographic and immunohistochemical analysis. Neuroscience. 2000;100:381–91.PubMedCrossRefGoogle Scholar
  47. 47.
    D'Antoni S, Zambusi L, Codazzi F, Zacchetti D, Grohovaz F, Provini L, et al. Calcitonin gene-related peptide (CGRP) stimulates purkinje cell dendrite growth in culture. Neurochem Res. 2010;35:2135–43.PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media New York 2013

Authors and Affiliations

  • Sajedeh Eftekhari
    • 1
  • Christopher A. Salvatore
    • 2
  • Renee C. Gaspar
    • 3
  • Rhonda Roberts
    • 3
  • Stacey O’Malley
    • 4
  • Zhizhen Zeng
    • 4
  • Lars Edvinsson
    • 1
  1. 1.Department of Clinical Sciences, Division of Experimental Vascular ResearchLund UniversityLundSweden
  2. 2.Department of Pain and Migraine ResearchMerck Research LaboratoriesWest PointUSA
  3. 3.Department of In Vivo PharmacologyMerck Research LaboratoriesWest PointUSA
  4. 4.Department of ImagingMerck Research LaboratoriesWest PointUSA

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