Korean J Physiol Pharmacol.  2018 Mar;22(2):173-182. 10.4196/kjpp.2018.22.2.173.

Oxytocin produces thermal analgesia via vasopressin-1a receptor by modulating TRPV1 and potassium conductance in the dorsal root ganglion neurons

Affiliations
  • 1Neuroscience Research Institute and Department of Physiology, Korea University College of Medicine, Seoul 02841, Korea. hsna@korea.ac.kr
  • 2Department of Brain and Cognitive Sciences, College of Natural Sciences, Seoul National University, Seoul 03080, Korea. odolbae@snu.ac.kr
  • 3Dental Research Institute and Department of Neurobiology & Physiology, School of Dentistry, Seoul National University, Seoul 08826, Korea.
  • 4Department of Physiology, College of Medicine, Gachon University, Incheon 21936, Korea.
  • 5Department of Neuroscience and Oral Physiology, Osaka University Graduate School of Dentistry, Osaka 565-0871, Japan.

Abstract

Recent studies have provided several lines of evidence that peripheral administration of oxytocin induces analgesia in human and rodents. However, the exact underlying mechanism of analgesia still remains elusive. In the present study, we aimed to identify which receptor could mediate the analgesic effect of intraperitoneal injection of oxytocin and its cellular mechanisms in thermal pain behavior. We found that oxytocin-induced analgesia could be reversed by d(CHâ‚‚)â‚…[Tyr(Me)²,Dab⁵] AVP, a vasopressin-1a (V1a) receptor antagonist, but not by desGly-NHâ‚‚-d(CHâ‚‚)â‚…[DTyr², Thr⁴]OVT, an oxytocin receptor antagonist. Single cell RT-PCR analysis revealed that V1a receptor, compared to oxytocin, vasopressin-1b and vasopressin-2 receptors, was more profoundly expressed in dorsal root ganglion (DRG) neurons and the expression of V1a receptor was predominant in transient receptor potential vanilloid 1 (TRPV1)-expressing DRG neurons. Fura-2 based calcium imaging experiments showed that capsaicin-induced calcium transient was significantly inhibited by oxytocin and that such inhibition was reversed by V1a receptor antagonist. Additionally, whole cell patch clamp recording demonstrated that oxytocin significantly increased potassium conductance via V1a receptor in DRG neurons. Taken together, our findings suggest that analgesic effects produced by peripheral administration of oxytocin were attributable to the activation of V1a receptor, resulting in reduction of TRPV1 activity and enhancement of potassium conductance in DRG neurons.

Keyword

Dorsal root ganglion; Electrophysiology; Oxytocin; Pain; Vasopressin receptor

MeSH Terms

Analgesia*
Calcium
Diagnosis-Related Groups
Electrophysiology
Fura-2
Ganglia, Spinal*
Humans
Injections, Intraperitoneal
Neurons
Oxytocin*
Potassium*
Receptors, Oxytocin
Receptors, Vasopressin
Rodentia
Spinal Nerve Roots*
Calcium
Fura-2
Oxytocin
Potassium
Receptors, Oxytocin
Receptors, Vasopressin

Figure

  • Fig. 1 Reversal of oxytocin-induced thermal analgesia via V1a receptor, not oxytocin receptor. (A) Thermal paw-withdrawal latency was measured and compared before the injection with 30 min and 60 min after the IP injection of vehicle, OT (1 mg/kg) and OT with OTRA (A selective oxytocin receptor antagonist 1 mg/kg), atosiban (an oxytocin and vasopressin receptor antagonist, 1 mg/kg) and V1aRA (a selective vasopressin 1a receptor antagonist, 1 mg/kg) in Hargreaves' test. **p<0.001, ***p<0.0001, Two-way RM ANOVA followed by posthoc Bonferroni's test. Error bars represent SEM (n=6, each group). (B) Nocifensive responses at 60 min after IP co-injection of antagonists were compared to the OT injection. **p<0.001, ***p<0.0001, One-way ANOVA followed by posthoc Bonferroni's test. Error bars represent SEM. (n=6, each group). OT, oxytocin; V1aRA, vasopressin-1a receptor antagonist; OTRA, oxytocin receptor antagonist; ns, not significant.

  • Fig. 2 The expression patterns of oxytocin receptors and TRPV1 in DRG neurons revealed by single cell RT-PCR analysis. (A) Representative gels show expression patterns of oxytocin binding receptors and TRPV1 from seven individual DRG neurons. Predicted size for selected markers are oxytocin receptor (158 bp), V1a receptor (151 bp), V1b receptor (194 bp), V2 receptor (V2, 153 bp), TRPV1 (330 bp) and GAPDH (268 bp). (B) The percentage of oxytocin binding receptors-positive cells in 54 GAPDH-positive DRG neurons. (C) The percentage of V1a receptor-positive cells within TRPV1 expressing DRG neurons. MM, molecular marker; NC, negative control.

  • Fig. 3 Oxytocin-induced decrease in capsaicin-induced calcium transients in the DRG neurons mediated via V1a receptor. Representative traces of intracellular calcium responses of cultured sensory neurons to 3 repetitive application of (A) 200 nM CAP for 10 s, (B) repetitive CAP application with pre-application of 5 µM of oxytocin for 120 s, (C) CAP application with 5 µM of oxytocin mixed with 20 µM of atosiban, and (D) 5 µM of oxytocin mixed with 5 µM of V1aRA before the second capsaicin application. (E) The proportion of oxytocin responsive cells within the DRG neurons responding to capsaicin in calcium imaging. (F) Summary of normalized ratio of capsaicin responses by without OT (n=21), OT alone (n=70), and OT with its antagonists, ATO (n=66) and V1aRA (n=70), relative to peak amplitude of 1st CAP transient. Before CAP application, 10 µM PMA was mixed with CAP solution. *p<0.05, One-way ANOVA followed by posthoc Bonferroni's test. Error bars represent SEM. CAP, capsaicin; PMA, Phorbol 12-myristate 13-acetate; OT, oxytocin; ATO, atosiban; V1aRA, vasopressin-1a receptor antagonist; ns, not significant.

  • Fig. 4 Decrease of action potential firings by oxytocin in small to medium-sized DRG neurons mediated via V1a receptor. (A, left panel) Representative recording illustrating effect of 5 µM oxytocin on action potential firing. The effect of oxytocin was reversed after 5-min washout. (A, right panel) Oxytocin decreased the mean number of action potential evoked by current injection (n=12). (B) 20 µM of Atosiban (n=9) and (C) 5 µM of V1aRA (n=9) reversed oxytocin-induced reduction of the number of action potentials in small to medium-sized DRG neurons. *p<0.01, ***p<0.0001, repeated measured-ANOVA followed by posthoc Bonferroni's test. Error bars represent SEM. OT, Oxytocin; V1aRA, vasopressin-1a receptor antagonist; ns, not significant.

  • Fig. 5 Enhancement of outward potassium currents by oxytocin in small to medium-sized DRG neurons mediated via V1a receptor. (A) Depolarizing (from –110 mV to 0 mV) ramp protocol for voltage-clamp experiments (B, left panel) Representative trace showing effects of oxytocin (5 µM) on I-V relationship response to ramp depolarization. (B, middle panel) Representative trace illustrating markedly enhanced a voltage-activated outward current and slightly enhanced a hyperpolarization-activated inward current (B, right panel) Oxytocin increased the mean peak current density at 0 mV by inducing outward potassium current in small to medium-sized DRG neurons (n=20). (C) 20 µM of atosiban (n=24) and (D) 5 µM of V1aRA blocked oxytocin-induced the voltage-activated outward potassium current and the hyperpolarization-activated inward current. TTX (0.5 µM) were treated in solution of all experiments (n=13). *p<0.01, **p<0.001, repeated measured-ANOVA followed by posthoc Bonferroni's test. Error bars represent SEM. OT, Oxytocin; V1aRA, vasopressin-1a receptor antagonist; ns, not significant.


Reference

1. Vrachnis N, Malamas FM, Sifakis S, Deligeoroglou E, Iliodromiti Z. The oxytocin-oxytocin receptor system and its antagonists as tocolytic agents. Int J Endocrinol. 2011; 2011:350546.
Article
2. González-Hernández A, Rojas-Piloni G, Condés-Lara M. Oxytocin and analgesia: future trends. Trends Pharmacol Sci. 2014; 35:549–551.
Article
3. Yang J. Intrathecal administration of oxytocin induces analgesia in low back pain involving the endogenous opiate peptide system. Spine (Phila Pa 1976). 1994; 19:867–871.
Article
4. Louvel D, Delvaux M, Felez A, Fioramonti J, Bueno L, Lazorthes Y, Frexinos J. Oxytocin increases thresholds of colonic visceral perception in patients with irritable bowel syndrome. Gut. 1996; 39:741–747.
Article
5. Lundeberg T, Uvnäs-Moberg K, Agren G, Bruzelius G. Anti-nociceptive effects of oxytocin in rats and mice. Neurosci Lett. 1994; 170:153–157.
Article
6. Kang YS, Park JH. Brain uptake and the analgesic effect of oxytocin−its usefulness as an analgesic agent. Arch Pharm Res. 2000; 23:391–395.
7. Juif PE, Poisbeau P. Neurohormonal effects of oxytocin and vasopressin receptor agonists on spinal pain processing in male rats. Pain. 2013; 154:1449–1456.
Article
8. Breton JD, Veinante P, Uhl-Bronner S, Vergnano AM, Freund-Mercier MJ, Schlichter R, Poisbeau P. Oxytocin-induced antinociception in the spinal cord is mediated by a subpopulation of glutamatergic neurons in lamina I-II which amplify GABAergic inhibition. Mol Pain. 2008; 4:19.
Article
9. Breton JD, Poisbeau P, Darbon P. Antinociceptive action of oxytocin involves inhibition of potassium channel currents in lamina II neurons of the rat spinal cord. Mol Pain. 2009; 5:63.
Article
10. Wrobel L, Schorscher-Petcu A, Dupré A, Yoshida M, Nishimori K, Tribollet E. Distribution and identity of neurons expressing the oxytocin receptor in the mouse spinal cord. Neurosci Lett. 2011; 495:49–54.
Article
11. Schorscher-Petcu A, Sotocinal S, Ciura S, Dupré A, Ritchie J, Sorge RE, Crawley JN, Hu SB, Nishimori K, Young LJ, Tribollet E, Quirion R, Mogil JS. Oxytocin-induced analgesia and scratching are mediated by the vasopressin-1A receptor in the mouse. J Neurosci. 2010; 30:8274–8284.
Article
12. Hobo S, Hayashida K, Eisenach JC. Oxytocin inhibits the membrane depolarization-induced increase in intracellular calcium in capsaicin sensitive sensory neurons: a peripheral mechanism of analgesic action. Anesth Analg. 2012; 114:442–449.
13. Qiu F, Qiu CY, Cai H, Liu TT, Qu ZW, Yang Z, Li JD, Zhou QY, Hu WP. Oxytocin inhibits the activity of acid-sensing ion channels through the vasopressin, V1A receptor in primary sensory neurons. Br J Pharmacol. 2014; 171:3065–3076.
14. Hargreaves K, Dubner R, Brown F, Flores C, Joris J. A new and sensitive method for measuring thermal nociception in cutaneous hyperalgesia. Pain. 1988; 32:77–88.
Article
15. Han RT, Lee H, Lee J, Lee SB, Kim HJ, Back SK, Na HS. Brief isolation changes nociceptive behaviors and compromises drug tests in mice. Pain Pract. 2016; 16:749–757.
Article
16. Kim YH, Park CK, Back SK, Lee CJ, Hwang SJ, Bae YC, Na HS, Kim JS, Jung SJ, Oh SB. Membrane-delimited coupling of TRPV1 and mGluR5 on presynaptic terminals of nociceptive neurons. J Neurosci. 2009; 29:10000–10009.
Article
17. Kim YB, Kim YS, Kim WB, Shen FY, Lee SW, Chung HJ, Kim JS, Han HC, Colwell CS, Kim YI. GABAergic excitation of vasopressin neurons: possible mechanism underlying sodium-dependent hypertension. Circ Res. 2013; 113:1296–1307.
18. Achilles K, Okabe A, Ikeda M, Shimizu-Okabe C, Yamada J, Fukuda A, Luhmann HJ, Kilb W. Kinetic properties of Cl uptake mediated by Na+-dependent K+-2Cl cotransport in immature rat neocortical neurons. J Neurosci. 2007; 27:8616–8627.
19. Díaz D, Bartolo R, Delgadillo DM, Higueldo F, Gomora JC. Contrasting effects of Cd2+ and Co2+ on the blocking/unblocking of human Cav3 channels. J Membr Biol. 2005; 207:91–105.
20. Mandadi S, Numazaki M, Tominaga M, Bhat MB, Armati PJ, Roufogalis BD. Activation of protein kinase C reverses capsaicin-induced calcium-dependent desensitization of TRPV1 ion channels. Cell Calcium. 2004; 35:471–478.
Article
21. Caterina MJ, Leffler A, Malmberg AB, Martin WJ, Trafton J, Petersen-Zeitz KR, Koltzenburg M, Basbaum AI, Julius D. Impaired nociception and pain sensation in mice lacking the capsaicin receptor. Science. 2000; 288:306–313.
Article
22. Manning M, Stoev S, Cheng LL, Wo NC, Chan WY. Design of oxytocin antagonists, which are more selective than atosiban. J Pept Sci. 2001; 7:449–465.
Article
23. Eliava M, Melchior M, Knobloch-Bollmann HS, Wahis J, da Silva Gouveia M, Tang Y, Ciobanu AC, Triana Del Rio R, Roth LC, Althammer F, Chavant V, Goumon Y, Gruber T, Petit-Demoulière N, Busnelli M, Chini B, Tan LL, Mitre M, Froemke RC, Chao MV, Giese G, Sprengel R, Kuner R, Poisbeau P, Seeburg PH, Stoop R, Charlet A, Grinevich V. A new population of parvocellular oxytocin neurons controlling magnocellular neuron activity and inflammatory pain processing. Neuron. 2016; 89:1291–1304.
Article
24. Chini B, Manning M. Agonist selectivity in the oxytocin/vasopressin receptor family: new insights and challenges. Biochem Soc Trans. 2007; 35:737–741.
Article
25. Dubin AE, Patapoutian A. Nociceptors: the sensors of the pain pathway. J Clin Invest. 2010; 120:3760–3772.
Article
26. Tan CH, McNaughton PA. The TRPM2 ion channel is required for sensitivity to warmth. Nature. 2016; 536:460–463.
Article
27. González-Hernández A, Manzano-García A, Martínez-Lorenzana G, Tello-García IA, Carranza M, Arámburo C, Condés-Lara M. Peripheral oxytocin receptors inhibit the nociceptive input signal to spinal dorsal horn wide-dynamic-range neurons. Pain. 2017; 158:2117–2128.
Article
28. Zhang H, Cang CL, Kawasaki Y, Liang LL, Zhang YQ, Ji RR, Zhao ZQ. Neurokinin-1 receptor enhances TRPV1 activity in primary sensory neurons via PKCepsilon: a novel pathway for heat hyperalgesia. J Neurosci. 2007; 27:12067–12077.
29. Moriyama T, Higashi T, Togashi K, Iida T, Segi E, Sugimoto Y, Tominaga T, Narumiya S, Tominaga M. Sensitization of TRPV1 by EP1 and IP reveals peripheral nociceptive mechanism of prostaglandins. Mol Pain. 2005; 1:3.
30. Premkumar LS, Ahern GP. Induction of vanilloid receptor channel activity by protein kinase C. Nature. 2000; 408:985–990.
Article
31. Bang S, Yoo S, Yang TJ, Cho H, Kim YG, Hwang SW. Resolvin D1 attenuates activation of sensory transient receptor potential channels leading to multiple anti-nociception. Br J Pharmacol. 2010; 161:707–720.
Article
32. Caires R, Luis E, Taberner FJ, Fernandez-Ballester G, Ferrer-Montiel A, Balazs EA, Gomis A, Belmonte C, de la Peña E. Hyaluronan modulates TRPV1 channel opening, reducing peripheral nociceptor activity and pain. Nat Commun. 2015; 6:8095.
Article
33. Neves SR, Ram PT, Iyengar R. G protein pathways. Science. 2002; 296:1636–1639.
Article
34. Terrillon S, Barberis C, Bouvier M. Heterodimerization of V1a and V2 vasopressin receptors determines the interaction with beta-arrestin and their trafficking patterns. Proc Natl Acad Sci U S A. 2004; 101:1548–1553.
35. Por ED, Bierbower SM, Berg KA, Gomez R, Akopian AN, Wetsel WC, Jeske NA. β-Arrestin-2 desensitizes the transient receptor potential vanilloid 1 (TRPV1) channel. J Biol Chem. 2012; 287:37552–37563.
Article
36. Gurevich VV. Arrestins-pharmacology and therapeutic potential. Heidelberg: Springer;2014.
37. Everill B, Rizzo MA, Kocsis JD. Morphologically identified cutaneous afferent DRG neurons express three different potassium currents in varying proportions. J Neurophysiol. 1998; 79:1814–1824.
Article
38. Gold MS, Shuster MJ, Levine JD. Characterization of six voltage-gated K+ currents in adult rat sensory neurons. J Neurophysiol. 1996; 75:2629–2646.
39. Du X, Gamper N. Potassium channels in peripheral pain pathways: expression, function and therapeutic potential. Curr Neuropharmacol. 2013; 11:621–640.
Article
40. Tsantoulas C, McMahon SB. Opening paths to novel analgesics: the role of potassium channels in chronic pain. Trends Neurosci. 2014; 37:146–158.
Article
41. Duan KZ, Xu Q, Zhang XM, Zhao ZQ, Mei YA, Zhang YQ. Targeting A-type K+ channels in primary sensory neurons for bone cancer pain in a rat model. Pain. 2012; 153:562–574.
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