Reduction of TRPV1 expression in the trigeminal system by botulinum neurotoxin type-A

Toshihiko Shimizu, Mamoru Shibata, Haruki Toriumi, Tatsuo Iwashita, Megumi Funakubo, Hitoshi Sato, Toshiya Kuroi, Taeko Ebine, Kenzo Koizumi, Norihiro Suzuki

Research output: Contribution to journalArticle

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Abstract

Botulinum neurotoxin type-A (BoNT-A) is clinically used for patients with pain disorders and dystonia. The precise mechanism whereby BoNT-A controls pain remains elusive. Here, we studied how BoNT-A affects the expression of the transient receptor potential vanilloid subfamily member 1 (TRPV1), a cation channel critically implicated in nociception, in the trigeminal system. Histological studies revealed that subcutaneous BoNT-A injection (0.25, 0.5, or 5. ng/kg) into the face targeted the ophthalmic division of trigeminal ganglion (TG) neurons and decreased TRPV1-immunoreactive neurons in the TG and TRPV1-immunoreactive fibers in rat trigeminal terminals. Of note, TG neurons that received projections from the dura mater, a principal site of headache generation, had reduced TRPV1 expression. BoNT-A-induced cleavage of SNAP25 (synaptosomal-associated protein of 25-kDa) in the TG became obvious 2. days after BoNT-A administration and persisted for at least 14. days. Quantitative real-time RT-PCR (reverse transcription-polymerase chain reaction) data indicated that the TRPV1-decreasing effects of BoNT-A were not mediated by transcriptional downregulation. By employing a surface protein biotin-labeling assay, we demonstrated that BoNT-A inhibited TRPV1 trafficking to the plasma membrane in primary TG neurons. Moreover, Y200F-mutated TRPV1, which is incapable of trafficking to the plasma membrane, was expressed in PC12 cells by transfection, and pharmacological studies revealed that TRPV1 in the cytoplasm was more predisposed to proteasome-mediated proteolysis than plasma membrane-located TRPV1. We conclude that the mechanism by which BoNT-A reduces TRPV1 expression involves the inhibition of TRPV1 plasma membrane trafficking and proteasome-mediated degradation in the cytoplasm. This paradigm seems to explain how BoNT-A alleviates TRPV1-mediated pain. Our data reveal a likely molecular mechanism whereby BoNT-A treatment reduces TRPV1 expression in the trigeminal system and provide important clues to novel therapeutic measures for ameliorating craniofacial pain.

Original languageEnglish
Pages (from-to)367-378
Number of pages12
JournalNeurobiology of Disease
Volume48
Issue number3
DOIs
Publication statusPublished - 2012 Dec

Fingerprint

TRPV Cation Channels
Type A Botulinum Toxins
Trigeminal Ganglion
Cell Membrane
Neurons
Proteasome Endopeptidase Complex
Cytoplasm
Synaptosomal-Associated Protein 25
Pain
Dura Mater
Facial Pain
Somatoform Disorders
Nociception
Dystonia
PC12 Cells
Biotin
Proteolysis
Reverse Transcription
Transfection
Headache

Keywords

  • Botulinum neurotoxin type-A
  • Craniofacial pain
  • Exocytosis
  • Migraine
  • Trigeminal ganglion
  • TRPV1

ASJC Scopus subject areas

  • Neurology

Cite this

Reduction of TRPV1 expression in the trigeminal system by botulinum neurotoxin type-A. / Shimizu, Toshihiko; Shibata, Mamoru; Toriumi, Haruki; Iwashita, Tatsuo; Funakubo, Megumi; Sato, Hitoshi; Kuroi, Toshiya; Ebine, Taeko; Koizumi, Kenzo; Suzuki, Norihiro.

In: Neurobiology of Disease, Vol. 48, No. 3, 12.2012, p. 367-378.

Research output: Contribution to journalArticle

Shimizu, T, Shibata, M, Toriumi, H, Iwashita, T, Funakubo, M, Sato, H, Kuroi, T, Ebine, T, Koizumi, K & Suzuki, N 2012, 'Reduction of TRPV1 expression in the trigeminal system by botulinum neurotoxin type-A', Neurobiology of Disease, vol. 48, no. 3, pp. 367-378. https://doi.org/10.1016/j.nbd.2012.07.010
Shimizu, Toshihiko ; Shibata, Mamoru ; Toriumi, Haruki ; Iwashita, Tatsuo ; Funakubo, Megumi ; Sato, Hitoshi ; Kuroi, Toshiya ; Ebine, Taeko ; Koizumi, Kenzo ; Suzuki, Norihiro. / Reduction of TRPV1 expression in the trigeminal system by botulinum neurotoxin type-A. In: Neurobiology of Disease. 2012 ; Vol. 48, No. 3. pp. 367-378.
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