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Identification of a Ca2+-Sensing Receptor in Rat Trigeminal Ganglia, Sensory Axons, and Tooth Dental Pulp

Calcified Tissue International volume 82, pages 57–65 (2008)Cite this article

Abstract

Extracellular Ca2+ regulates dentin formation, but little information is available on this regulatory mechanism. We have previously reported that sensory denervation reduces dentin formation, suggesting a role for sensory nerves in tooth mineralization. The G protein-coupled Ca2+-sensing receptor (CaR) is expressed in dorsal root ganglia and perivascular sensory nerves in mesenteric arterioles, and activation of these receptors by Ca2+ has been shown to induce vascular relaxation. The present study determined CaR expression in tooth dental pulp (DP), sensory axons, and trigeminal ganglion (TG) as well as the effect of increased [Ca2+]e or a calcimimetic on tooth blood flow. The distribution of CaR, studied by immunochemistry, RT-PCR, and Western blot, indicates abundant expression of CaR in sensory axons in the jaws, TG, and DP. Restriction analysis of PCR products with specific endonucleases showed the presence of CaR message in TG and DP, and Western blotting indicates the expression of mature and immature forms of the receptor in these tissues. Pulpal blood flow, measured by laser-Doppler flowmetry, increased by 67% ± 6% (n = 12) following receptor stimulation with 5 mM Ca2+, which was completely inhibited by 5 μM IBTx, a high-conductance KCa channel blocker indicating a mechanism involving hyperpolarization. NPS R-467 (10 μM) increased blood flow by 85% ± 18% (n = 6), suggesting regulation through the CaR. Our results suggest that the CaR is present in sensory nerves, DP, and TG and that an increase in Ca2+ in the DP causes vasodilatation, which may contribute to accumulation of Ca2+ during dentin mineralization.

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References

  1. Brown EM, Gamba G, Riccardi D, Lombardi M, Butters R, Kifor O, Sun A, Hediger MA, Lytton J, Hebert SC (1993) Cloning and characterization of an extracellular Ca(2+)-sensing receptor from bovine parathyroid. Nature 366:575–580

    PubMed  Article  CAS  Google Scholar 

  2. Riccardi D, Park J, Lee WS, Gamba G, Brown EM, Hebert SC (1995) Cloning and functional expression of a rat kidney extracellular calcium/polyvalent cation-sensing receptor. Proc Natl Acad Sci USA 92:131–135

    PubMed  Article  CAS  Google Scholar 

  3. Ruat M, Molliver ME, Snowman AM, Snyder SH (1995) Calcium sensing receptor: molecular cloning in rat and localization to nerve terminals. Proc Natl Acad Sci USA 92:3161–3165

    PubMed  Article  CAS  Google Scholar 

  4. Bukoski RD, Bian K, Wang Y, Mupanomunda M (1997) Perivascular sensory nerve Ca2+ receptor and Ca2+-induced relaxation of isolated arteries. Hypertension 30:1431–1439

    PubMed  CAS  Google Scholar 

  5. Wang Y, Awumey EK, Chatterjee PK, Somasundaram C, Bian K, Rogers KV, Dunn C, Bukoski RD (2003) Molecular cloning and characterization of a rat sensory nerve Ca2+-sensing receptor. Am J Physiol Cell Physiol 285:C64–C75

    PubMed  CAS  Google Scholar 

  6. Bukoski RD (1998) The perivascular sensory nerve Ca2+ receptor and blood pressure regulation: a hypothesis. Am J Hypertens 11:1117–1123

    PubMed  Article  CAS  Google Scholar 

  7. Mupanomunda MM, Tian B, Ishioka N, Bukoski RD (2000) Renal interstitial Ca(2+). Am J Physiol Renal Physiol 278:F644–F649

    PubMed  CAS  Google Scholar 

  8. Bukoski RD, Batkai S, Jarai Z, Wang Y, Offertaler L, Jackson WF, Kunos G (2002) CB(1) receptor antagonist SR141716A inhibits Ca(2+)-induced relaxation in CB(1) receptor-deficient mice. Hypertension 39:251–257

    PubMed  Article  CAS  Google Scholar 

  9. Awumey EM, Howlett AC, Putney JW Jr, Diz DI, Bukoski RD (2007) Ca(2+) mobilization through dorsal root ganglion Ca(2+)-sensing receptor stably expressed in HEK293 cells. Am J Physiol Cell Physiol 292:C1895–C1905

    PubMed  Article  CAS  Google Scholar 

  10. Heyeraas KJ, Kvinnsland I, Byers MR, Jacobsen EB (1993) Nerve fibers immunoreactive to protein gene product 9.5, calcitonin gene-related peptide, substance P, and neuropeptide Y in the dental pulp, periodontal ligament, and gingiva in cats. Acta Odontol Scand 51:207–221

    PubMed  Article  CAS  Google Scholar 

  11. Vongsavan N, Matthews B (1992) Changes in pulpal blood flow and in fluid flow through dentine produced by autonomic and sensory nerve stimulation in the cat. Proc Finn Dent Soc 88(Suppl 1):491–497

    PubMed  Google Scholar 

  12. Kerezoudis NP, Olgart L, Edwall L (1993) Differential effects of nitric oxide synthesis inhibition on basal blood flow and antidromic vasodilation in rat oral tissues. Eur J Pharmacol 241:209–219

    PubMed  Article  CAS  Google Scholar 

  13. Berggreen E, Heyeraas KJ (1999) The role of sensory neuropeptides and nitric oxide on pulpal blood flow and tissue pressure in the ferret. J Dent Res 78:1535–1543

    PubMed  CAS  Article  Google Scholar 

  14. Jacobsen EB, Heyeraas KJ (1996) Effect of capsaicin treatment or inferior alveolar nerve resection on dentine formation and calcitonin gene-related peptide- and substance P-immunoreactive nerve fibres in rat molar pulp. Arch Oral Biol 41:1121–1131

    PubMed  Article  CAS  Google Scholar 

  15. Haug SR, Berggreen E, Heyeraas KJ (2001) The effect of unilateral sympathectomy and cavity preparation on peptidergic nerves and immune cells in rat dental pulp. Exp Neurol 169:182–190

    PubMed  Article  CAS  Google Scholar 

  16. Price TJ, Louria MD, Candelario-Soto D, Dussor GO, Jeske NA, Patwardhan AM, Diogenes A, Trott AA, Hargreaves KM, Flores CM (2005) Treatment of trigeminal ganglion neurons in vitro with NGF, GDNF or BDNF: effects on neuronal survival, neurochemical properties and TRPV1-mediated neuropeptide secretion. BMC Neurosci 6:4

    PubMed  Article  CAS  Google Scholar 

  17. Mathias RS, Mathews CH, Machule C, Gao D, Li W, Denbesten PK (2001) Identification of the calcium-sensing receptor in the developing tooth organ. J Bone Miner Res 16:2238–2244

    PubMed  Article  CAS  Google Scholar 

  18. Ishioka N, Bukoski RD (1999) A role for N-arachidonylethanolamine (anandamide) as the mediator of sensory nerve-dependent Ca2+-induced relaxation. J Pharmacol Exp Ther 289:245–250

    PubMed  CAS  Google Scholar 

  19. Furuta T, Shimizu Y, Takahashi I, Mitani H (1999) Formation and mineralization of murine molar roots with hypocalcaemia induced by a low-calcium diet and the changes after returning to a normal diet. Arch Oral Biol 44:629–639

    PubMed  Article  CAS  Google Scholar 

  20. Lundgren T, Nannmark U, Linde A (1992) Calcium ion activity and pH in the odontoblast-predentin region: ion-selective microelectrode measurements. Calcif Tissue Int 50:134–136

    PubMed  Article  CAS  Google Scholar 

  21. Nicholson WA, Ashton BA, Hohling HJ, Quint P, Schreiber J, Ashton IK, Boyde A (1977) Electron microprobe investigations into the process of hard tissue formation. Cell Tissue Res 177:331–345

    PubMed  Article  CAS  Google Scholar 

  22. Lundquist P (2002) Odontoblast phosphate and calcium transport in dentinogenesis. Swed Dent J Suppl:1–52

    Google Scholar 

  23. Linde A (1995) Dentin mineralization and the role of odontoblasts in calcium transport. Connect Tissue Res 33:163–170

    PubMed  Article  CAS  Google Scholar 

  24. Lundgren T, Linde A (1997) Voltage-gated calcium channels and nonvoltage-gated calcium uptake pathways in the rat incisor odontoblast plasma membrane. Calcif Tissue Int 60:79–85

    PubMed  Article  CAS  Google Scholar 

  25. Linde A, Lundgren T (1995) From serum to the mineral phase. The role of the odontoblast in calcium transport and mineral formation. Int J Dev Biol 39:213–222

    PubMed  CAS  Google Scholar 

  26. Datta HK, Horrocks BR (2003) Mechanisms of calcium disposal from osteoclastic resorption hemivacuole. J Endocrinol 176:1–5

    PubMed  Article  CAS  Google Scholar 

  27. Silver IA, Murrills RJ, Etherington DJ (1988) Microelectrode studies on the acid microenvironment beneath adherent macrophages and osteoclasts. Exp Cell Res 175:266–276

    PubMed  Article  CAS  Google Scholar 

  28. Fan G, Goldsmith PK, Collins R, Dunn CK, Krapcho KJ, Rogers KV, Spiegel AM (1997) N-linked glycosylation of the human Ca2 + receptor is essential for its expression at the cell surface. Endocrinology 138:1916–1922

    PubMed  Article  CAS  Google Scholar 

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Acknowledgments

This work was supported by Western Norway Regional Health Authority Fund and Faculty of Dentistry Norwegian Research Council Project 101330 and NIH Grants UH1 HL059868, R25 HL059868-08, and R01 HL064761.

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Affiliations

  1. Department of Biomedicine, Section for Physiology, Faculty of Medicine, University of Bergen, Jonas Lies vei 91, N-5009, Bergen, Norway

    Karin J. Heyeraas & Sivakami R. Haug

  2. Julius L Chambers Biomedical/Biotechnology Research Institute, North Carolina Central University, Durham, NC, 27707, USA

    Richard D. Bukoski & Emmanuel M. Awumey

Authors
  1. Karin J. Heyeraas
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  2. Sivakami R. Haug
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  3. Richard D. Bukoski
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  4. Emmanuel M. Awumey
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Correspondence to Karin J. Heyeraas.

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Heyeraas, K.J., Haug, S.R., Bukoski, R.D. et al. Identification of a Ca2+-Sensing Receptor in Rat Trigeminal Ganglia, Sensory Axons, and Tooth Dental Pulp. Calcif Tissue Int 82, 57–65 (2008). https://doi.org/10.1007/s00223-007-9096-z

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  • DOI: https://doi.org/10.1007/s00223-007-9096-z

Keywords

  • Sensory nerve
  • Calcium-sensing receptor
  • Vasodilatation
  • Teeth
  • Mineralization