Spinal TNF-α impedes Fbxo45-dependent Munc13-1 ubiquitination to mediate neuropathic allodynia in rats

Ming Chun Hsieh, Yu Cheng Ho, Cheng Yuan Lai, Dylan Chou, Gin Den Chen, Tzer Bin Lin, Hsien Yu Peng

Research output: Contribution to journalArticle

3 Citations (Scopus)

Abstract

Presynaptic active zone proteins play a crucial role in regulating synaptic plasticity. Although the ubiquitin–proteasome system underlying the degradation of the presynaptic active zone protein is well established, the contribution of this machinery to regulating spinal plasticity during neuropathic pain development remains unclear. Here, using male Sprague Dawley rats, we demonstrated along with behavioral allodynia, neuropathic injury induced a marked elevation in the expression levels of an active zone protein Munc13-1 in the homogenate and synaptic plasma membrane of the ipsilateral dorsal horn. Moreover, nerve injury-increased Munc13-1 expression was associated with an increase in the frequency and amplitude of miniature excitatory postsynaptic currents (mEPSCs) in ipsilateral dorsal horn neurons. This neuropathic injury-induced accumulation of Munc13-1 colocalized with synaptophysin but not homer1 in the dorsal horn. Focal knockdown of spinal Munc13-1 expression attenuated behavioral allodynia and the increased frequency, not the amplitude, of mEPSCs in neuropathic rats. Remarkably, neuropathic injury decreased spinal Fbxo45 expression, Fbxo45-Munc13-1 co-precipitation, and Munc13-1 ubiquitination in the ipsilateral dorsal horn. Conversely, focal knockdown of spinal Fbxo45 expression in naive animals resulted in behavioral allodynia in association with similar protein expression and ubiquitination in the dorsal horn as observed with neuropathic injury rats. Furthermore, both neuropathic insults and intrathecal injection of tumor necrosis factor-α (TNF-α) impeded spinal Fbxo45-dependent Munc13-1 ubiquitination, which was reversed by intrathecal TNF-α-neutralizing antibody. Our data revealed that spinal TNF-α impedes Fbxo45-dependent Munc13-1 ubiquitination that accumulates Munc13-1 in the presynaptic area and hence facilitates the synaptic excitability of nociceptive neurotransmission underlying neuropathic pain.

Original languageEnglish
Article number811
JournalCell Death and Disease
Volume9
Issue number8
DOIs
Publication statusPublished - Aug 1 2018

Fingerprint

Ubiquitination
Hyperalgesia
Tumor Necrosis Factor-alpha
Excitatory Postsynaptic Potentials
Wounds and Injuries
Neuralgia
Proteins
Posterior Horn Cells
Spinal Injections
Spinal Injuries
Synaptic Membranes
Synaptophysin
Neuronal Plasticity
Neutralizing Antibodies
Synaptic Transmission
Sprague Dawley Rats
Cell Membrane
Spinal Cord Dorsal Horn

ASJC Scopus subject areas

  • Immunology
  • Cellular and Molecular Neuroscience
  • Cell Biology
  • Cancer Research

Cite this

Spinal TNF-α impedes Fbxo45-dependent Munc13-1 ubiquitination to mediate neuropathic allodynia in rats. / Hsieh, Ming Chun; Ho, Yu Cheng; Lai, Cheng Yuan; Chou, Dylan; Chen, Gin Den; Lin, Tzer Bin; Peng, Hsien Yu.

In: Cell Death and Disease, Vol. 9, No. 8, 811, 01.08.2018.

Research output: Contribution to journalArticle

Hsieh, Ming Chun ; Ho, Yu Cheng ; Lai, Cheng Yuan ; Chou, Dylan ; Chen, Gin Den ; Lin, Tzer Bin ; Peng, Hsien Yu. / Spinal TNF-α impedes Fbxo45-dependent Munc13-1 ubiquitination to mediate neuropathic allodynia in rats. In: Cell Death and Disease. 2018 ; Vol. 9, No. 8.
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abstract = "Presynaptic active zone proteins play a crucial role in regulating synaptic plasticity. Although the ubiquitin–proteasome system underlying the degradation of the presynaptic active zone protein is well established, the contribution of this machinery to regulating spinal plasticity during neuropathic pain development remains unclear. Here, using male Sprague Dawley rats, we demonstrated along with behavioral allodynia, neuropathic injury induced a marked elevation in the expression levels of an active zone protein Munc13-1 in the homogenate and synaptic plasma membrane of the ipsilateral dorsal horn. Moreover, nerve injury-increased Munc13-1 expression was associated with an increase in the frequency and amplitude of miniature excitatory postsynaptic currents (mEPSCs) in ipsilateral dorsal horn neurons. This neuropathic injury-induced accumulation of Munc13-1 colocalized with synaptophysin but not homer1 in the dorsal horn. Focal knockdown of spinal Munc13-1 expression attenuated behavioral allodynia and the increased frequency, not the amplitude, of mEPSCs in neuropathic rats. Remarkably, neuropathic injury decreased spinal Fbxo45 expression, Fbxo45-Munc13-1 co-precipitation, and Munc13-1 ubiquitination in the ipsilateral dorsal horn. Conversely, focal knockdown of spinal Fbxo45 expression in naive animals resulted in behavioral allodynia in association with similar protein expression and ubiquitination in the dorsal horn as observed with neuropathic injury rats. Furthermore, both neuropathic insults and intrathecal injection of tumor necrosis factor-α (TNF-α) impeded spinal Fbxo45-dependent Munc13-1 ubiquitination, which was reversed by intrathecal TNF-α-neutralizing antibody. Our data revealed that spinal TNF-α impedes Fbxo45-dependent Munc13-1 ubiquitination that accumulates Munc13-1 in the presynaptic area and hence facilitates the synaptic excitability of nociceptive neurotransmission underlying neuropathic pain.",
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