Skip to main content

Advertisement

Log in

The activation of transient receptor potential vanilloid receptor subtype 1 by capsaicin without extracellular Ca2+ is involved in the mechanism of distinct substance P release in cultured rat dorsal root ganglion neurons

  • Original Article
  • Published:
Naunyn-Schmiedeberg's Archives of Pharmacology Aims and scope Submit manuscript

Abstract

Using a highly sensitive substance P (SP) radioimmunoassay, we attempted to elucidate the molecular mechanism of SP release from cultured adult rat dorsal root ganglion (DRG) neurons triggered by capsaicin. As a result, we found that capsaicin can induce SP release in the absence of extracellular Ca2+ by activating transient receptor potential vanilloid receptor subtype 1 (TRPV1). Therefore, we compared the pharmacological profile of SP release involved in several intracellular effectors (phosphoinositide 3-kinase (PI3K), Ca2+ release from intracellular stores, and mitogen-activated protein (MAP) kinases) in the presence/absence of extracellular Ca2+ by stimulating DRG neurons with various concentrations (10 to 1,000 nM) of capsaicin. In the presence of extracellular Ca2+, a capsaicin-induced maximal release of SP obtained at 100 nM capsaicin was attenuated by either the phosphoinositide 3-kinase (PI3K) inhibitors (wortmannin and LY294002), the inositol 1,4,5-triphosphate (IP3)-induced Ca2+ release blocker (2-aminoethyl diphenylborinate, 2-APB) or a specific antagonist of TRPV1 (capsazepine). In contrast, in the absence of extracellular Ca2+, only a high concentration (1 μM) of capsaicin induced a significant increase in the SP release, which was then completely abolished by either a mitogen-activated protein kinase kinase (MEK) inhibitor U0126 or capsazepine, and significantly inhibited by either thapsigargin (a Ca2+-ATPase inhibitor) or BAPTA-AM (a rapid Ca2+ chelator). In summary, the activation of TRPV1 by capsaicin modulates the SP release from DRG neurons via two different mechanisms, one requiring extracellular Ca2+, the activation of PI3K and the IP3-dependent intracellular Ca2+ release, and the other which is independent of extracellular Ca2+ but involves the activation of MEK. These data suggest that a distinct SP release mechanism exists on DRG through the activation of TRPV1 without extracellular Ca2+ by capsaicin to facilitate neuropeptide release.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5

Similar content being viewed by others

References

  • Ayar A, Scott RH (1998) The actions of ryanodine on Ca2+-activated conductances in rat cultured DRG neurons; evidence for Ca2+-induced Ca2+ release. Naunyn-Schmiedeberg’s Arch Pharmacol 359:81–91

    Article  Google Scholar 

  • Barendrecht MM, Mulders AC, van der Poel H, van den Hoff MJ, Schmidt M, Michel MC (2007) Role of transforming growth factor beta in rat bladder smooth muscle cell proliferation. J Pharmacol Exp Ther 322:117–122

    Article  PubMed  CAS  Google Scholar 

  • Bito H, Deisseroth K, Tsien RW (1996) CREB phosphorylation and dephosphorylation: a Ca2+- and stimulus duration-dependent switch for hippocampal gene expression. Cell 87:1203–1214

    Article  PubMed  CAS  Google Scholar 

  • Caterina MJ, Julius D (2001) The vanilloid receptor: a molecular gateway to the pain pathway. Annu Rev Neurosci 24:487–517

    Article  PubMed  CAS  Google Scholar 

  • Dai Y, Iwata K, Fukuoka T, Kondo E, Tokunaga A, Yamanaka H, Tachibana T, Liu Y, Noguchi K (2002) Phosphorylation of extracellular signal-regulated kinase in primary afferent neurons by noxious stimuli and its involvement in peripheral sensitization. J Neurosci 22:7737–7745

    PubMed  CAS  Google Scholar 

  • Dinh QT, Groneberg DA, Peiser C, Mingomataj E, Joachim RA, Witt C, Arck PC, Klapp BF, Fischer A (2004) Substance P expression in TRPV1 and trkA-positive dorsal root ganglion neurons innervating the mouse lung. Respir Physiol Neurobiol 144:15–24

    Article  PubMed  CAS  Google Scholar 

  • Eun SY, Jung SJ, Park YK, Kwak J, Kim SJ, Kim J (2001) Effects of capsaicin on Ca2+ release from the intracellular Ca2+ stores in the dorsal root ganglion cells of adult rats. Biochem Biophys Res Commun 285:1114–1120

    Article  PubMed  CAS  Google Scholar 

  • Grady EF, Yoshimi SK, Maa J, Valeroso D, Vartanian RK, Rahim S, Kim EH, Gerard C, Gerard N, Bunnett NW, Kirkwood KS (2000) Substance P mediates inflammatory oedema in acute pancreatitis via activation of the neurokinin-1 receptor in rats and mice. Br J Pharmacol 130:505–512

    Article  PubMed  CAS  Google Scholar 

  • Gu Q, Lin RL, Hu HZ, Zhu MX, Lee LY (2005) 2-aminoethoxydiphenyl borate stimulates pulmonary C neurons via the activation of TRPV channels. Am J Physiol Lung Cell Mol Physiol 288:932–941

    Article  Google Scholar 

  • Helliwell RJ, McLatchie LM, Clarke M, Winter J, Bevan S, McIntyre P (1998) Capsaicin sensitivity is associated with the expression of the vanilloid (capsaicin) receptor (VR1) mRNA in adult rat sensory ganglia. Neurosci Lett 250:177–180

    Article  PubMed  CAS  Google Scholar 

  • Jin SX, Zhuang ZY, Woolf CJ, Ji RR (2003) p38 mitogen-activated protein kinase is activated after a spinal nerve ligation in spinal cord microglia and dorsal root ganglion neurons and contributes to the generation of neuropathic pain. J Neurosci 23:4017–4022

    PubMed  CAS  Google Scholar 

  • Kárai LJ, Russell JT, Iadarola MJ, Oláh Z (2004) Vanilloid receptor 1 regulates multiple calcium compartments and contributes to Ca2+-induced Ca2+ release in sensory neurons. J Biol Chem 279:16377–16387

    Article  PubMed  Google Scholar 

  • Marshall IC, Owen DE, Cripps TV, Davis JB, McNulty S, Smart D (2003) Activation of vanilloid receptor 1 by resiniferatoxin mobilizes calcium from inositol 1,4,5-trisphosphate-sensitive stores. Br J Pharmacol 138:172–176

    Article  PubMed  CAS  Google Scholar 

  • Numazaki M, Tominaga T, Takeuchi K, Murayama N, Toyooka H, Tominaga M (2003) Structural determinant of TRPV1 desensitization interacts with calmodulin. Proc Natl Acad Sci USA 100:8002–8006

    Article  PubMed  CAS  Google Scholar 

  • Oláh Z, Szabo T, Kárai L, Hough C, Fields RD, Caudle RM, Blumberg PM, Iadarola MJ (2001) Ligand-induced dynamic membrane changes and cell deletion conferred by vanilloid receptor 1. J Biol Chem 276:11021–11030

    Article  PubMed  Google Scholar 

  • Pearson G, Robinson F, Beers Gibson T, Xu BE, Karandikar M, Berman K, Cobb MH (2001) Mitogen-activated protein (MAP) kinase pathways: regulation and physiological functions. Endocr Rev 22:153–183

    Article  PubMed  CAS  Google Scholar 

  • Price TJ, Flores CM (2007) Critical evaluation of the colocalization between calcitonin gene-related peptide, SP, transient receptor potential vanilloid subfamily type 1 immunoreactivities, and isolectin B(4) binding in primary afferent neurons of the rat and mouse. J Pain 8:263–272

    Article  PubMed  CAS  Google Scholar 

  • Purkiss J, Welch M, Doward S, Foster K (2000) Capsaicin-stimulated release of substance P from cultured dorsal root ganglion neurons: involvement of two distinct mechanisms. Biochem Pharmacol 59:1403–1406

    Article  PubMed  CAS  Google Scholar 

  • Severini C, Improta G, Falconieri-Erspamer G, Salvadori S, Erspamer V (2002) The tachykinin peptide family. Pharmacological Reviews 54:285–322

    Article  PubMed  CAS  Google Scholar 

  • Stutzmann GE (2005) Calcium dysregulation, IP3 signaling, and Alzheimer’s disease. Neuroscientist 11:110–115

    Article  PubMed  CAS  Google Scholar 

  • Suzuki H, Yoshioka K, Maehara T, Guo JZ, Nonomura Y, Otsuka M (1998) Differential effects of wortmannin on the release of SP and amino acids from the isolated spinal cord of the neonatal rat. Br J Pharmacol 125:1661–1668

    Article  PubMed  CAS  Google Scholar 

  • Suzuki T, Hide I, Ido K, Kohsaka S, Inoue K, Nakata Y (2004) Production and release of neuroprotective tumor necrosis factor by P2X7 receptor-activated microglia. J Neurosci 24:1–7

    Article  PubMed  CAS  Google Scholar 

  • Suzuki T, Hide I, Matsubara A, Hama C, Harada K, Miyano K, Andrä M, Matsubayashi H, Sakai N, Kohsaka S, Inoue K, Nakata Y (2006) Microglial alpha7 nicotinic acetylcholine receptors drive a phospholipase C/IP3 pathway and modulate the cell activation toward a neuroprotective role. J Neurosci Res 83:1461–1470

    Article  PubMed  CAS  Google Scholar 

  • Szallasi A (2006) Small molecule vanilloid TRPV1 receptor antagonists approaching drug status: can they live up to the expectations? Naunyn Schmiedebergs Arch Pharmacol 373:273–286

    Article  PubMed  CAS  Google Scholar 

  • Tan M, Groszer M, Tan AM, Pandya A, Liu X, Xie CW (2003) Phosphoinositide 3-kinase cascade facilitates μ-opioid desensitization in sensory neurons by altering G-protein–effector interactions. J Neurosci 23:10292–10301

    PubMed  CAS  Google Scholar 

  • Tang HB, Nakata Y (2006) Olopatadine attenuates the enhancement of capsaicin-evoked SP release by bradykinin from cultured dorsal root ganglion neurons. Eur J Pharmacol 552:78–82

    Article  PubMed  CAS  Google Scholar 

  • Tang HB, Inoue A, Oshita K, Nakata Y (2004) Sensitization of vanilloid receptor 1 induced by bradykinin via the activation of second messenger signaling cascades in rat primary afferent neurons. Eur J Pharmacol 498:37–43

    Article  PubMed  CAS  Google Scholar 

  • Tang HB, Inoue A, Oshita K, Hirate K, Nakata Y (2005) Zaltoprofen inhibits bradykinin-induced responses by blocking the activation of second messenger signaling cascades in rat dorsal root ganglion cells. Neuropharmacology 48:1035–1042

    Article  PubMed  CAS  Google Scholar 

  • Tang HB, Inoue A, Iwasa M, Hide I, Nakata Y (2006) SP release evoked by capsaicin or potassium from rat cultured dorsal root ganglion neurons is conversely modulated with bradykinin. J Neurochem 97:1412–1418

    Article  PubMed  CAS  Google Scholar 

  • Toker A, Cantley LC (1997) Signalling through the lipid products of phosphoinositide-3-OH kinase. Nature 387:673–676

    Article  PubMed  CAS  Google Scholar 

  • Tsuzuki K, Xing H, Ling J, Gu JG (2004) Menthol-induced Ca2+ release from presynaptic Ca2+ stores potentiates sensory synaptic transmission. J Neurosci 24:762–771

    Article  PubMed  CAS  Google Scholar 

  • Van der Stelt M, Trevisani M, Vellani V, De Petrocellis L, Schiano Moriello A, Campi B, McNaughton P, Geppetti P, Di Marzo V (2005) Anandamide acts as an intracellular messenger amplifying Ca2+ influx via TRPV1 channels. EMBO J 24:3026–3037

    Article  PubMed  Google Scholar 

  • Zhuang ZY, Xu H, Clapham DE, Ji RR (2004) Phosphatidylinositol 3-kinase activates ERK in primary sensory neurons and mediates inflammatory heat hyperalgesia through TRPV1 sensitization. J Neurosci 24:8300–8309

    Article  PubMed  CAS  Google Scholar 

  • Zhuang ZY, Wen YR, Zhang DR, Borsello T, Bonny C, Strichartz GR, Decosterd I, Ji RR (2006) A peptide c-Jun N-terminal kinase (JNK) inhibitor blocks mechanical allodynia after spinal nerve ligation: respective roles of JNK activation in primary sensory neurons and spinal astrocytes for neuropathic pain development and maintenance. J Neurosci 26:3551–3560

    Article  PubMed  CAS  Google Scholar 

Download references

Acknowledgments

This study was supported by a Grant-in-Aid for Scientific Research (B) from the Japan Society for the Promotion of Science (JSPS) to H.B. Tang (No. 19790189). We thank Dr. Brian Quinn (Kyushu University, Japan) for his critical reading of the manuscript.

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Yoshihiro Nakata.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Tang, HB., Nakata, Y. The activation of transient receptor potential vanilloid receptor subtype 1 by capsaicin without extracellular Ca2+ is involved in the mechanism of distinct substance P release in cultured rat dorsal root ganglion neurons. Naunyn-Schmied Arch Pharmacol 377, 325–332 (2008). https://doi.org/10.1007/s00210-007-0211-5

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00210-007-0211-5

Keywords

Navigation