Mitochondrial calcium uniporter contributes to morphine tolerance through pCREB and CPEB1 in rat spinal cord dorsal horn

K. Takahashi, H. Yi, J. Gu, D. Ikegami, S. Liu, T. Iida, Y. Kashiwagi, C. Dong, T. Kunisawa, Shuanglin Hao

Research output: Contribution to journalArticle

Abstract

Background: The long-term use of opioid analgesics is limited by the development of unwanted side-effects, such as tolerance. The molecular mechanisms of morphine anti-nociceptive tolerance are still unclear. The mitochondrial calcium uniporter (MCU) is involved in painful hyperalgesia, but the role of MCU in morphine tolerance has not been uncharacterised. Methods: Rats received intrathecal injection of morphine for 7 days to induce morphine tolerance. The mechanical withdrawal threshold was measured using von Frey filaments, and thermal latency using the hotplate test. The effects of an MCU inhibitor, antisense oligodeoxynucleotide against cyclic adenosine monophosphate response element (CRE)-binding protein (CREB) or cytoplasmic polyadenylation element-binding protein 1 (CPEB1) in morphine tolerance were examined. Results: Spinal morphine tolerance was associated with an increased expression of neuronal MCU, phospho-CREB (pCREB), and CPEB1 in the spinal cord dorsal horn. MCU inhibition increased the mechanical threshold and thermal latency, and reduced the accumulation of mitochondrial calcium in morphine tolerance. Intrathecal antisense oligodeoxynucleotide against CREB or CPEB1 restored the anti-nociceptive effects of morphine compared with mismatch oligodeoxynucleotide in von Frey test and hotplate test. Chromatin immunoprecipitation with quantitative PCR assay showed that CREB knockdown reduced the interaction of pCREB with the ccdc109a gene (encoding MCU expression) promoter and decreased the MCU mRNA transcription. RNA immunoprecipitation assay suggested that CPEB1 binds to the MCU mRNA 3′ untranslated region. CPEB1 knockdown decreased the expression of MCU protein. Conclusions: These findings suggest that spinal MCU is regulated by pCREB and CPEB1 in morphine tolerance, and that inhibition of MCU, pCREB, or CPEB1 may be useful in preventing the development of opioid tolerance.

Original languageEnglish (US)
JournalBritish Journal of Anaesthesia
DOIs
StatePublished - Jan 1 2019

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Polyadenylation
Morphine
Carrier Proteins
Oligodeoxyribonucleotides
Opioid Analgesics
Spinal Cord Dorsal Horn
mitochondrial calcium uniporter
Hot Temperature
Spinal Injections
Messenger RNA
Chromatin Immunoprecipitation
Hyperalgesia
Response Elements
3' Untranslated Regions
Immunoprecipitation
Cyclic AMP
RNA

Keywords

  • calcium
  • CPEB
  • CREB
  • mitochondrial calcium uniporter
  • morphine
  • opioid tolerance
  • opioids

ASJC Scopus subject areas

  • Anesthesiology and Pain Medicine

Cite this

Mitochondrial calcium uniporter contributes to morphine tolerance through pCREB and CPEB1 in rat spinal cord dorsal horn. / Takahashi, K.; Yi, H.; Gu, J.; Ikegami, D.; Liu, S.; Iida, T.; Kashiwagi, Y.; Dong, C.; Kunisawa, T.; Hao, Shuanglin.

In: British Journal of Anaesthesia, 01.01.2019.

Research output: Contribution to journalArticle

Takahashi, K. ; Yi, H. ; Gu, J. ; Ikegami, D. ; Liu, S. ; Iida, T. ; Kashiwagi, Y. ; Dong, C. ; Kunisawa, T. ; Hao, Shuanglin. / Mitochondrial calcium uniporter contributes to morphine tolerance through pCREB and CPEB1 in rat spinal cord dorsal horn. In: British Journal of Anaesthesia. 2019.
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AU - Yi, H.

AU - Gu, J.

AU - Ikegami, D.

AU - Liu, S.

AU - Iida, T.

AU - Kashiwagi, Y.

AU - Dong, C.

AU - Kunisawa, T.

AU - Hao, Shuanglin

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AB - Background: The long-term use of opioid analgesics is limited by the development of unwanted side-effects, such as tolerance. The molecular mechanisms of morphine anti-nociceptive tolerance are still unclear. The mitochondrial calcium uniporter (MCU) is involved in painful hyperalgesia, but the role of MCU in morphine tolerance has not been uncharacterised. Methods: Rats received intrathecal injection of morphine for 7 days to induce morphine tolerance. The mechanical withdrawal threshold was measured using von Frey filaments, and thermal latency using the hotplate test. The effects of an MCU inhibitor, antisense oligodeoxynucleotide against cyclic adenosine monophosphate response element (CRE)-binding protein (CREB) or cytoplasmic polyadenylation element-binding protein 1 (CPEB1) in morphine tolerance were examined. Results: Spinal morphine tolerance was associated with an increased expression of neuronal MCU, phospho-CREB (pCREB), and CPEB1 in the spinal cord dorsal horn. MCU inhibition increased the mechanical threshold and thermal latency, and reduced the accumulation of mitochondrial calcium in morphine tolerance. Intrathecal antisense oligodeoxynucleotide against CREB or CPEB1 restored the anti-nociceptive effects of morphine compared with mismatch oligodeoxynucleotide in von Frey test and hotplate test. Chromatin immunoprecipitation with quantitative PCR assay showed that CREB knockdown reduced the interaction of pCREB with the ccdc109a gene (encoding MCU expression) promoter and decreased the MCU mRNA transcription. RNA immunoprecipitation assay suggested that CPEB1 binds to the MCU mRNA 3′ untranslated region. CPEB1 knockdown decreased the expression of MCU protein. Conclusions: These findings suggest that spinal MCU is regulated by pCREB and CPEB1 in morphine tolerance, and that inhibition of MCU, pCREB, or CPEB1 may be useful in preventing the development of opioid tolerance.

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