Skip to main content
SpringerLink
Log in

[3H]resiniferatoxin binding by the vanilloid receptor: species-related differences, effects of temperature and sulfhydryl reagents

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

Summary

Specific binding of [3H]resiniferatoxin (RTX) is thought to represent the vanilloid (capsaicin) receptor. In the present study, we have used this binding assay to elucidate the contribution of differential receptor expression to the capsaicin-resistance of hamsters and rabbits; binding parameters were compared to those of species (rats, mice) regarded as capsaicin-sensitive. Whereas the 5-fold lower affinity for [3H]RTX binding in the hamster (100 pM) as compared to the rat (20 pM) is unlikely to account for the 100-fold difference in the in vivo responses of RTX-induced inflammation and hypothermia, the lack of detectable specific [3H]RTX binding sites in the rabbit might represent the predominant mechanism of capsaicin-resistance in this species. Regulation of the vanilloid receptor was further characterized in the rat. In accord with the temperature dependence of both in vivo and in vitro capsaicin actions, we found a marked temperature dependence for association rates. Dissociation turned out to have complex kinetics dependent on time and receptor occupancy. Low pH (5.5–7.0) did not affect receptor binding. Preincubation with heavy metal cations and other sulfhydryl-reactive agents inhibited specific [3H]RTX binding indicating that the vanilloid receptor is a thiol-protein, and that free sulfhydryl groups play an essential role in agonist binding activity. Preliminary characterization suggested noncompetitive inhibition.

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

Access this article

Log in via an institution

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

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  • Amann R (1990) Desensitization of capsaicin-evoked neuropeptide release — Influence of Ca2+ and temperature. Naunyn-Schmiedeberg's Arch Pharmacol 342:671–676

    Google Scholar 

  • Aronstam RS, Abood LG, Hoss W (1978) Influence of sulfhydryl reagents and heavy metals on the functional state of the muscarinic acetylcholine receptor in rat brain. Mol Pharmacol 14:575- 586

    Google Scholar 

  • Baranowski R, Lynn B, Pini A (1986) The effects of locally applied capsaicin on conduction in cutaneous nerves in four mammalian species. Br J Pharmacol 89:267–276

    Google Scholar 

  • Baumann TK, Simone DA, Shain CN, LaMotte RH (1991) Neurogenic hyperalgesia: the search for the primary cutaneous afferent fibers that contribute to capsaicin-induced pain and hyperalgesia. J Neurophysiol 66:212–227

    CAS  PubMed  Google Scholar 

  • Bevan S, Yeats J (1991) Protons activate a cation conductance in a subpopulation of rat dorsal root ganglion neurones. J Physiol (Lond) 433:145–161

    Google Scholar 

  • Bevan S, James IF, Rang HP, Winter J, Wood JN (1987) The mechanism of action of capsaicin — a sensory neurotoxin. In: Jenner P (ed) Neurotoxins and their pharmacological implications. Raven Press, New York, pp 261–277

    Google Scholar 

  • Braestrup C, Andersen PH (1987) Effects of heavy metal cations and other sulfhydryl reagents on brain dopamine D1 receptors: evidence for involvement of a thiol group in the conformation of the active site. J Neurochem 48:1667–1672

    Google Scholar 

  • Buck SH, Burks TIT (1986) The neuropharmacology of capsaicin: review of some recent observations. Pharmacol Rev 38:179–226

    Google Scholar 

  • Buck SH, Walsh JH, Davis TP, Brown MR, Yamamura HI, Burks TF (1983) Characterization of the peptide and sensory neurotoxic effect of capsaicin in the guinea pig. J Neurosci 3:2064–2074

    Google Scholar 

  • Buckley TL, Brain SD, Williams TJ (1990) Ruthenium red selectively inhibits oedema formation and increased blood flow induced by capsaicin in rabbit skin. Br J Pharmacol 99:7–8

    Google Scholar 

  • Carter RB (1991) Topical capsaicin in the treatment of cutaneous disorders. Drug Development Res 22:109–123

    Google Scholar 

  • DeVries DJ, Blumberg PM (1989) Thermoregulatory effects of resiniferatoxin in the mouse: comparison with capsaicin. Life Sci 44:711–715

    Google Scholar 

  • Docherty RJ, Robertson B, Bevan S (1991) Capsaicin causes prolonged inhibition of voltage-activated calcium currents in adult rat dorsal root ganglion neurons in culture. Neuroscience 40:513–521

    Google Scholar 

  • Dray A, Hankins MW, Yeats JC (1989) Desensitization and capsaicin-induced release of substance P-like immunoreactivity from guinea-pig ureter in vitro. Neuroscience 31:479–483

    Google Scholar 

  • Endrenyi L, Fajszi CS, Kwong FHF (1975) Evaluation of Hill slopes and Hill coefficients when the saturation binding or velocity is not known. Eur J Biochem 51:317–328

    Google Scholar 

  • Evangelista S, Renzi D, Guzzi P, Surrenti C, Santicioli P, Maggi CA (1992) Capsaicin-like activity of N-ethylmaleimide in rat stomach. Gen Pharmac 23:39–41

    Google Scholar 

  • Geppetti P, Theodorsson-Norheim E, Ballerini G, Alessandri M, Maggi CA, Santicioli P, Amenta F, Fanciullacci M (1988) Capsaicin-sensitive tachykinin-like immunoreactivity in the thymus of rats and guinea-pigs. J Neuroimmunol 19:3–9

    Google Scholar 

  • Geppetti P, DelBianco E, Patacchini R, Santicioli P, Maggi CA, Tramontana M (1991) Low pH-induced release of calcitonin gene-related peptide from capsaicin-sensitive sensory nerves: mechanism of action and biological response. Neuroscience 41:295–301

    Google Scholar 

  • Glinsukon T, Stitmunnaithum V, Toskulkao C, Baranawuti T, Tangkrisanavinont V (1980) Acute toxicity of capsaicin in several animal species. Toxicon 18:215–220

    Google Scholar 

  • Guard S, Watson SP (1991) Tachykinin receptor subtypes: classification and membrane signalling mechanisms. Neurochem Int 18:149–165

    Google Scholar 

  • Hergenhahn M, Adolph W, Hecker E (1975) Resiniferatoxin and other esters of novel polyfunctional diterpenes from Euphorbia resinifera and unispina. Tetrahedron Lett 19:1595–1598

    Google Scholar 

  • Holzer P (1988) Local effector functions of capsaicin-sensitive sensory nerve endings: involvement of tachykinins, calcitonin gene-related peptide and other peptides. Neuroscience 24:739–768

    Google Scholar 

  • Holzer P (1991) Capsaicin: cellular targets, mechanisms of action, and selectivity for thin sensory neurons. Pharmacol Rev 43:144–201

    Google Scholar 

  • Jancso N (1960) Role of nerve terminals in the mechanism of inflammatory reactions. Bull Millard Fillmore Hosp (Buffalo, NY) 7:53–77

    Google Scholar 

  • Jancso N (1968) Desensitization with capsaicin and related acylamides as a tool for studying the function of pain receptors. In: Lin K, Armstrong D, Pardo KS (eds) Pharmacology of pain, vol 9. Pergamon Press, Oxford, pp 33–55

    Google Scholar 

  • Jancso G (1990) B-afferents: a system of capsaicin-sensitive primary sensory neurons. Behav Brain Sci 13:306–307

    Google Scholar 

  • Jancso N, Jancso-Labor A, Szolcsanyi J (1967) Direct evidence for neurogenic inflammation and its prevention by denervation and by pretreatment with capsaicin. Br J Pharmaco Chemother 31:138–151

    Google Scholar 

  • Joo F, Szolcsanyi J, Jancso-Labor A (1969) Mitochondrial alterations in the spinal ganglion cells of the rat accompanying the long-lasting sensory disturbance induced by capsaicin. Life Sci 8:621–626

    Google Scholar 

  • Kanui TI, Hole K, Miaron JO (1990) Nociception in crocodiles: capsaicin instillation, formalin and hot plate tests. Zool Sci 7:537–540

    Google Scholar 

  • Karlin A, Bartels E (1966) Effects of blocking sulfhydryl groups and of reducing disulfide bonds on the acetylcholine-activated permeability system of the electroplax. Biochim Biophys Acta 126:525–535

    Google Scholar 

  • Kiskin NI, Krishtal OA, Tsyndrenko AYA, Akaike N (1986) Are sulfhydryl groups essential for function of the glutamate-operated receptor-ionophore complex? Neurosci Lett 66:305–310

    Google Scholar 

  • Laitinen LA, Laitinen A, Panula PA, Partanen M, Tervo K, Tervo T (1983) Immunohistochemical demonstration of substance P in the lower respiratory tract of the rabbit and not in man. Thorax 38:531–536

    Google Scholar 

  • Lembeck F, Bucsics A (1990) Classification of peripheral neurons. Behav Brain Sci 13:310–311

    Google Scholar 

  • Maggi CA (1991) The pharmacology of the efferent functions of sensory nerves. J Autonom Pharmac 11:173–208

    Google Scholar 

  • Maggi CA, Meli A (1986) The role of neuropeptides in the regulation of the micturition reflex. J Autonom Pharmac 6:133–162

    Google Scholar 

  • Maggi CA, Meli A (1988) The sensory-efferent function of capsaicin-sensitive sensory neurons. Gen Pharmacol 19:1–43

    Google Scholar 

  • Maggi CA, Giuliani S, Santicioli P, Abelli L, Geppetti P, Somma V, Renzi D, Meli A (1987) Species-related variations in the effects of capsaicin on urinary bladder functions: relation to bladder content of substance P-like immunoreactivity. Naunyn-Schmiedeberg's Arch Pharmacol 336:546–555

    Google Scholar 

  • Miller MS, Buck SH, Sipes IG, Yamamura HI, Burks TF (1982) Regulation of substance P by nerve growth factor: disruption by capsaicin. Brain Res 250:193–196

    Google Scholar 

  • Rumsfield JA, West DP (1991) Topical capsaicin in dermatologic and peripheral pain disorders. DICP Ann Pharmacother 25:381–387

    Google Scholar 

  • Spina D, McKenniff MG, Coyle AJ, Seeds EAM, Tramontana M, Perretti F, Manzini S, Page CP (1991) Effect of capsaicin on PAF-induced bronchial hyperresponsiveness and pulmonary cell accumulation in the rabbit. Br J Pharmacol 103:1268–1274

    Google Scholar 

  • Szallasi A, Blumberg PM (1989) Resiniferatoxin, a phorbol-related diterpene, acts as an ultrapotent analog of capsaicin, the irritant constituent in red pepper. Neuroscience 30:515–520

    Google Scholar 

  • Szallasi A, Blumberg PM (1990a) Specific binding of resiniferatoxin, an ultrapotent capsaicin analog, by dorsal root ganglion membranes. Brain Res 524:106–111

    Google Scholar 

  • Szallasi A, Blumberg PM (1990b) Minireview: Resiniferatoxin and its analogs provide novel insights into the pharmacology of the vanilloid (capsaicin) receptor. Life Sci 47:1399–1408

    Google Scholar 

  • Szallasi A, Blumberg PM (1992a) Resiniferatoxin. In: Conn PM (ed) Methods in neuroscience, vol 8. Neurotoxins. Academic Press, Orlando, Fl., pp 368–380

    Google Scholar 

  • Szallasi A and Blumberg PM (1992b) Vanilloid receptor loss in rat sensory ganglia associated with long term desensitization to resiniferatoxin. Neurosci Lett 136:51–54

    Google Scholar 

  • Szallasi A, Lewin NE, Blumberg PM (1992) Identification of Alpha-1-acid glycoprotein (orosomucoid) as a major vanilloid binding protein in serum. J Pharmacol Exp Ther 262:883–888

    Google Scholar 

  • Szallasi A, Sharkey NA, Blumberg PM (1989) Structure-activity analysis of resiniferatoxin analogs. Phytother Res 3:253–257

    Google Scholar 

  • Szolcsanyi J (1977) A pharmacological approach to elucidation of the role of different nerve fibers and receptor endings in mediation of pain. J Physiol (Paris) 73:251–259

    Google Scholar 

  • Szolcsanyi J (1984) Capsaicin-sensitive chemoceptive neural system with dual sensory-efferent function. In: Chahl LA, Szolcsanyi J, Lembeck F (eds) Antidromic vasodilatation and neurogenic inflammation. Akademiai Kiado, Budapest, pp 27–52

    Google Scholar 

  • Szolcsanyi J, Joo F, Jancso-Gabor A (1971) Mitochondrial changes in preoptic neurones after capsaicin desensitization of the hypothalamic thermodetectors in the rat. Nature 229:116–117

    Google Scholar 

  • Wood JN, Winter J, James IF, Rang HP, Yeats J, Bevan S (1988) Capsaicin-induced ion fluxes in dorsal root glanglion cells in culture. J Neurosci 8:3208–3220

    Google Scholar 

Download references

Author information

Author notes

  1. Arpad Szallasi

    Present address: Department of Pharmacology, Menarini Ricerche Sud, Via Tito Speri 10, I-00040, Pomezia (Roma), Italy

Authors and Affiliations

  1. Molecular Mechanisms of Tumor Promotion Section, Laboratory of Cellular Carcinogenesis and Tumor Promotion, National Cancer Institute, 20892, Bethesda, MD, USA

    Arpad Szallasi & Peter M. Blumberg

Authors
  1. Arpad Szallasi
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Peter M. Blumberg
    View author publications

    You can also search for this author in PubMed Google Scholar

Additional information

Correspondence to P. M. Blumberg at the above address

Rights and permissions

Reprints and permissions

About this article

Cite this article

Szallasi, A., Blumberg, P.M. [3H]resiniferatoxin binding by the vanilloid receptor: species-related differences, effects of temperature and sulfhydryl reagents. Naunyn-Schmiedeberg's Arch Pharmacol 347, 84–91 (1993). https://doi.org/10.1007/BF00168777

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00168777

Key words

Search

Navigation