Contribution of transient receptor potential vanilloid subfamily 1 to endothelin-1-induced thermal hyperalgesia

doi:10.1016/j.neuroscience.2008.04.010

Neuroscience

Volume 154, Issue 3, 26 June 2008, Pages 1067-1076

https://doi.org/10.1016/j.neuroscience.2008.04.010 Get rights and content

Abstract

Endothelin-1 (ET-1) plays an important role in peripheral pain processing. However, the mechanisms of the nociceptive action of ET-1 have not been fully elucidated. In this study, we investigated the contribution of transient receptor potential vanilloid subfamily 1 (TRPV1) to ET-1-induced thermal hyperalgesia. Intraplantar ET-1-induced thermal hyperalgesia was examined by assessing the paw withdrawal latency to noxious heat stimuli. In electrophysiological study, whole-cell patch-clamp recordings were performed to investigate the interaction of ET-1 and TRPV1 using human embryonic kidney 293 (HEK293) cells expressing endothelin type A receptor (ET_A) and TRPV1. Intraplantar ET-1 (3, 10 and 30 pmol) produced thermal hyperalgesia in a dose-dependent manner. Thermal hyperalgesia was attenuated by the inhibition of ET_A and protein kinase C (PKC) but not that of ET_B. ET-1-induced thermal hyperalgesia was significantly attenuated in TRPV1-deficient mice compared with that in wild-type mice. In voltage-clamp experiments, 10 nM capsaicin evoked small inward currents in HEK293 cells expressing TRPV1 and ET_A. In the presence of ET-1, capsaicin produced much larger current responses (P<0.05). Mutation at PKC-specific TRPV1 phosphorylation sites (S800A/S502A) and PKC inhibitors inhibited the potentiating effect of ET-1. In addition, ET-1 decreased the temperature threshold for TRPV1 activation in a PKC-dependent manner (from 41.0±0.4 °C to 32.6±0.6 °C). In addition, Western blot analysis was also performed to confirm ET-1-induced phosphorylation of TRPV1. Incubation of ET-1 and intraplantar ET-1 evoked phosphorylation of TRPV1 in HEK293 cells expressing TRPV1 and ET_A and the skin, respectively. These results suggest that the sensitization of TRPV1 activity through an ET_A-PKC pathway contributes to ET-1-induced thermal hyperalgesia.

Section snippets

Animals

Male C57BL/6-strain mice (wild-type mice, WT mice) (weighing 20–25 g, Japan SLC; Hamamatsu, Shizuoka, Japan) and male TRPV1-deficient mice (TRPV1 KO mice) (weighing 20–25 g, Jackson Laboratory, Bar Harbor, MA, USA) were used for behavioral analyses. Each mouse was used in only one experiment. The mice were housed in a temperature-controlled (21±1 °C) room with a 12-h light/dark cycle and given free access to food and water. All procedures were approved by the Sapporo Medical University Animal

Intraplantar ET-1-induced thermal hyperalgesia in WT mice

Intraplantar ET-1 evoked thermal hyperalgesia as previously reported (Menendez et al., 2003). Although the vehicle did not affect the withdrawal latency, ET-1 (3, 10 and 30 pmol) decreased the withdrawal latencies in a dose-dependent manner (P<0.05) (Fig. 1A,Fig. 2B). Peak effects were observed 10–20 min after ET-1 injection, and the decreased withdrawal latencies returned to the basal withdrawal latencies within 60 min after injection. In order to clarify the involvement of receptor subtypes,

Discussion

Our major finding in the behavioral study was that the activation of PKC and TRPV1 as well as that of ET_A contributed to ET-1-induced thermal hyperalgesia. In addition, using a specific antibody, we obtained in vivo and in vitro evidence that ET-1 phosphorylated TRPV1. In an electrophysiological study, ET-1 enhanced TRPV1 responsiveness to capsaicin and decreased the temperature threshold of TRPV1 through PKC activation. Although a recent study using HEK293 cells expressing ET_A and TRPV1 or

Conclusion

Our results indicate that the sensitization of TRPV1 activity through an ET_A–PKC pathway contributes to ET-1-induced thermal hyperalgesia. Therefore, a strategy to target TRPV1 may be more effective to reduce pain associated with ET-1.

Acknowledgments

T.K. and J.Y. contributed equally to this work. Support was provided by a Grant-in-Aid for Scientific Research (19659403 and 19791076) to T. Kawamata and Y. Niiyama, respectively.

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Citation Excerpt :
In the dorsal root ganglia (DRG) neurons, it has been reported that ETA receptors are widely expressed in TRPV1-positive small diameter sensory neurons, while expression of ETB receptors is restricted to satellite glial cells (Plant et al., 2007; Plant, Zollner, Mousa, & Oksche, 2006; Pomonis, Rogers, Peters, Ghilardi, & Mantyh, 2001). ET-1 application promotes large increases in capsaicin-evoked currents on mice and rats DRG neurons, an effect that seems to be mediated by ETA receptors and to involve the activation of protein kinase C (PKC) (Kawamata et al., 2008; Plant et al., 2006; Plant et al., 2007). Corroborating these in vitro observations, it was reported that ET-1 induced heat hyperalgesia was attenuated in TRPV1-deficient mice (Ji et al., 2007; Kawamata et al., 2008).
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ET-1 (100 pmol/50 μL) was injected into the upper lip and heat hyperalgesia was evaluated for up to 6 h. Facial heat hyperalgesia induced by ET-1 was assessed in rats pre-treated locally with BQ-123 or BQ-788 (selective ET_A and ET_B receptor antagonists, respectively, 30 nmol/50 μL); BCTC (TRPV1 receptor antagonist; 300 μg/50 μL); anti-NGF (3 μg/50 μL); K252a (TrkA inhibitor, 1 μg/50 μL); or in rats that received intraganglionar resiniferatoxin injection (RTX, 200 ng/10 μL) to promote C-fibers ablation.
ET-1 induced facial heat hyperalgesia that persisted up to 6 h and was prevented by BQ-123, BQ-788 or by intraganglionar RTX injection. Likewise, local pre-treatment with BCTC abolished ET-1 induced facial heat hyperalgesia up to 3 h. Local pre-treatment with anti-NGF or K252a was effective to prevent ET-1 induced heat hyperalgesia.
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Pain mechanismContribution of transient receptor potential vanilloid subfamily 1 to endothelin-1-induced thermal hyperalgesia

Abstract

Section snippets

Animals

Intraplantar ET-1-induced thermal hyperalgesia in WT mice

Discussion

Conclusion

Acknowledgments

Exp Neurol

Pain

Life Sci

Eur J Pharmacol

Neuron

Pain

J Biol Chem

Neuroscience

Eur J Pharmacol

J Chem Neuroanat

Neuron

Neuroscience

Neuroscience

Involvement of endogenous endothelins in thermal and mechanical inflammatory hyperalgesia in mice

Naunyn Schmiedebergs Arch Pharmacol

Protein kinase C mediates up-regulation of tetrodotoxin-resistant, persistent Na+ current in rat and mouse sensory neurones

J Physiol

Protein kinase C phosphorylation sensitizes but does not activate the capsaicin receptor transient receptor potential vanilloid 1 (TRPV1)

Proc Natl Acad Sci U S A

Impaired nociception and pain sensation in mice lacking the capsaicin receptor

Science

Regional haemodynamic effects of endothelin-1 in rat and man: unexpected adverse reaction

J Hypertens

Pain mechanism
Contribution of transient receptor potential vanilloid subfamily 1 to endothelin-1-induced thermal hyperalgesia

Protein kinase C mediates up-regulation of tetrodotoxin-resistant, persistent Na⁺ current in rat and mouse sensory neurones