Skip to main content

Main menu

  • Home
  • Articles
    • Current
    • Special Feature Articles - Most Recent
    • Special Features
    • Colloquia
    • Collected Articles
    • PNAS Classics
    • List of Issues
  • Front Matter
    • Front Matter Portal
    • Journal Club
  • News
    • For the Press
    • This Week In PNAS
    • PNAS in the News
  • Podcasts
  • Authors
    • Information for Authors
    • Editorial and Journal Policies
    • Submission Procedures
    • Fees and Licenses
  • Submit
  • Submit
  • About
    • Editorial Board
    • PNAS Staff
    • FAQ
    • Accessibility Statement
    • Rights and Permissions
    • Site Map
  • Contact
  • Journal Club
  • Subscribe
    • Subscription Rates
    • Subscriptions FAQ
    • Open Access
    • Recommend PNAS to Your Librarian

User menu

  • Log in
  • My Cart

Search

  • Advanced search
Home
Home
  • Log in
  • My Cart

Advanced Search

  • Home
  • Articles
    • Current
    • Special Feature Articles - Most Recent
    • Special Features
    • Colloquia
    • Collected Articles
    • PNAS Classics
    • List of Issues
  • Front Matter
    • Front Matter Portal
    • Journal Club
  • News
    • For the Press
    • This Week In PNAS
    • PNAS in the News
  • Podcasts
  • Authors
    • Information for Authors
    • Editorial and Journal Policies
    • Submission Procedures
    • Fees and Licenses
  • Submit
Research Article

Morphine paradoxically prolongs neuropathic pain in rats by amplifying spinal NLRP3 inflammasome activation

View ORCID ProfilePeter M. Grace, Keith A. Strand, Erika L. Galer, Daniel J. Urban, Xiaohui Wang, Michael V. Baratta, Timothy J. Fabisiak, Nathan D. Anderson, Kejun Cheng, Lisa I. Greene, Debra Berkelhammer, Yingning Zhang, Amanda L. Ellis, Hang Hubert Yin, Serge Campeau, Kenner C. Rice, Bryan L. Roth, Steven F. Maier, and Linda R. Watkins
  1. aDepartment of Psychology and Neuroscience, University of Colorado, Boulder, CO 80309;
  2. bThe Center for Neuroscience, University of Colorado, Boulder, CO 80309;
  3. cDiscipline of Pharmacology, School of Medicine, University of Adelaide, Adelaide, SA 5005, Australia;
  4. dDepartment of Pharmacology, University of North Carolina, Chapel Hill, NC 27599;
  5. eChemical Biology Laboratory, Changchun Institute of Applied Chemistry, Chinese Academy of Sciences, Changchun, Jilin 130022, China;
  6. fDepartment of Chemistry and Biochemistry, University of Colorado, Boulder, CO 80309;
  7. gBioFrontiers Institute, University of Colorado, Boulder, CO 80309;
  8. hThe Center for Neuroscience, University of Colorado, Boulder, CO 80309;
  9. iChemical Biology Research Branch, National Institute on Drug Abuse and National Institute on Alcohol Abuse and Alcoholism, Bethesda, MD 20892;
  10. jCenter of Basic Molecular Science, Department of Chemistry, Tsinghua University, Beijing 100082, China

See allHide authors and affiliations

PNAS June 14, 2016 113 (24) E3441-E3450; first published May 31, 2016; https://doi.org/10.1073/pnas.1602070113
Peter M. Grace
aDepartment of Psychology and Neuroscience, University of Colorado, Boulder, CO 80309;
bThe Center for Neuroscience, University of Colorado, Boulder, CO 80309;
cDiscipline of Pharmacology, School of Medicine, University of Adelaide, Adelaide, SA 5005, Australia;
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
  • ORCID record for Peter M. Grace
  • For correspondence: peter.grace@colorado.edu
Keith A. Strand
aDepartment of Psychology and Neuroscience, University of Colorado, Boulder, CO 80309;
bThe Center for Neuroscience, University of Colorado, Boulder, CO 80309;
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Erika L. Galer
aDepartment of Psychology and Neuroscience, University of Colorado, Boulder, CO 80309;
bThe Center for Neuroscience, University of Colorado, Boulder, CO 80309;
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Daniel J. Urban
dDepartment of Pharmacology, University of North Carolina, Chapel Hill, NC 27599;
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Xiaohui Wang
aDepartment of Psychology and Neuroscience, University of Colorado, Boulder, CO 80309;
bThe Center for Neuroscience, University of Colorado, Boulder, CO 80309;
eChemical Biology Laboratory, Changchun Institute of Applied Chemistry, Chinese Academy of Sciences, Changchun, Jilin 130022, China;
fDepartment of Chemistry and Biochemistry, University of Colorado, Boulder, CO 80309;
gBioFrontiers Institute, University of Colorado, Boulder, CO 80309;
hThe Center for Neuroscience, University of Colorado, Boulder, CO 80309;
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Michael V. Baratta
aDepartment of Psychology and Neuroscience, University of Colorado, Boulder, CO 80309;
bThe Center for Neuroscience, University of Colorado, Boulder, CO 80309;
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Timothy J. Fabisiak
aDepartment of Psychology and Neuroscience, University of Colorado, Boulder, CO 80309;
bThe Center for Neuroscience, University of Colorado, Boulder, CO 80309;
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Nathan D. Anderson
aDepartment of Psychology and Neuroscience, University of Colorado, Boulder, CO 80309;
bThe Center for Neuroscience, University of Colorado, Boulder, CO 80309;
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Kejun Cheng
iChemical Biology Research Branch, National Institute on Drug Abuse and National Institute on Alcohol Abuse and Alcoholism, Bethesda, MD 20892;
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Lisa I. Greene
aDepartment of Psychology and Neuroscience, University of Colorado, Boulder, CO 80309;
bThe Center for Neuroscience, University of Colorado, Boulder, CO 80309;
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Debra Berkelhammer
aDepartment of Psychology and Neuroscience, University of Colorado, Boulder, CO 80309;
bThe Center for Neuroscience, University of Colorado, Boulder, CO 80309;
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Yingning Zhang
aDepartment of Psychology and Neuroscience, University of Colorado, Boulder, CO 80309;
bThe Center for Neuroscience, University of Colorado, Boulder, CO 80309;
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Amanda L. Ellis
aDepartment of Psychology and Neuroscience, University of Colorado, Boulder, CO 80309;
bThe Center for Neuroscience, University of Colorado, Boulder, CO 80309;
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Hang Hubert Yin
fDepartment of Chemistry and Biochemistry, University of Colorado, Boulder, CO 80309;
gBioFrontiers Institute, University of Colorado, Boulder, CO 80309;
hThe Center for Neuroscience, University of Colorado, Boulder, CO 80309;
jCenter of Basic Molecular Science, Department of Chemistry, Tsinghua University, Beijing 100082, China
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Serge Campeau
aDepartment of Psychology and Neuroscience, University of Colorado, Boulder, CO 80309;
bThe Center for Neuroscience, University of Colorado, Boulder, CO 80309;
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Kenner C. Rice
iChemical Biology Research Branch, National Institute on Drug Abuse and National Institute on Alcohol Abuse and Alcoholism, Bethesda, MD 20892;
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Bryan L. Roth
dDepartment of Pharmacology, University of North Carolina, Chapel Hill, NC 27599;
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Steven F. Maier
aDepartment of Psychology and Neuroscience, University of Colorado, Boulder, CO 80309;
bThe Center for Neuroscience, University of Colorado, Boulder, CO 80309;
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Linda R. Watkins
aDepartment of Psychology and Neuroscience, University of Colorado, Boulder, CO 80309;
bThe Center for Neuroscience, University of Colorado, Boulder, CO 80309;
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
  1. Edited by David Julius, University of California, San Francisco, CA, and approved April 19, 2016 (received for review February 16, 2016)

  • Article
  • Figures & SI
  • Info & Metrics
  • PDF
Loading

Significance

Pain after disease/damage of the nervous system is predominantly treated with opioids, but without exploration of the long-term consequences. We demonstrate that a short course of morphine after nerve injury doubles the duration of neuropathic pain. Using genetic and pharmacological interventions, and innovative Designer Receptor Exclusively Activated by Designer Drugs disruption of microglia reactivity, we demonstrate that opioid-prolonged neuropathic pain arises from spinal microglia and NOD-like receptor protein 3 inflammasome formation/activation. Inhibiting these processes permanently resets amplified pain to basal levels, an effect not previously reported. These data support the “two-hit hypothesis” of amplification of microglial activation—nerve injury being the first “hit,” morphine the second. The implications of such potent microglial “priming” has fundamental clinical implications for pain and may extend to many chronic neurological disorders.

Abstract

Opioid use for pain management has dramatically increased, with little assessment of potential pathophysiological consequences for the primary pain condition. Here, a short course of morphine, starting 10 d after injury in male rats, paradoxically and remarkably doubled the duration of chronic constriction injury (CCI)-allodynia, months after morphine ceased. No such effect of opioids on neuropathic pain has previously been reported. Using pharmacologic and genetic approaches, we discovered that the initiation and maintenance of this multimonth prolongation of neuropathic pain was mediated by a previously unidentified mechanism for spinal cord and pain—namely, morphine-induced spinal NOD-like receptor protein 3 (NLRP3) inflammasomes and associated release of interleukin-1β (IL-1β). As spinal dorsal horn microglia expressed this signaling platform, these cells were selectively inhibited in vivo after transfection with a novel Designer Receptor Exclusively Activated by Designer Drugs (DREADD). Multiday treatment with the DREADD-specific ligand clozapine-N-oxide prevented and enduringly reversed morphine-induced persistent sensitization for weeks to months after cessation of clozapine-N-oxide. These data demonstrate both the critical importance of microglia and that maintenance of chronic pain created by early exposure to opioids can be disrupted, resetting pain to normal. These data also provide strong support for the recent “two-hit hypothesis” of microglial priming, leading to exaggerated reactivity after the second challenge, documented here in the context of nerve injury followed by morphine. This study predicts that prolonged pain is an unrealized and clinically concerning consequence of the abundant use of opioids in chronic pain.

  • TLR4
  • P2X7R
  • danger signals
  • DAMP
  • opioid-induced hyperalgesia

Footnotes

  • ↵1To whom correspondence should be addressed. Email: peter.grace{at}colorado.edu.
  • Author contributions: P.M.G., X.W., M.V.B., H.H.Y., S.F.M., and L.R.W. designed research; P.M.G., K.A.S., E.L.G., X.W., M.V.B., T.J.F., N.D.A., L.I.G., D.B., Y.Z., A.L.E., and S.C. performed research; D.J.U., K.C., S.C., K.C.R., and B.L.R. contributed new reagents/analytic tools; P.M.G. analyzed data; and P.M.G., X.W., S.C., S.F.M., and L.R.W. wrote the paper.

  • The authors declare no conflict of interest.

  • This article is a PNAS Direct Submission.

  • This article contains supporting information online at www.pnas.org/lookup/suppl/doi:10.1073/pnas.1602070113/-/DCSupplemental.

View Full Text
PreviousNext
Back to top
Article Alerts
Email Article

Thank you for your interest in spreading the word on PNAS.

NOTE: We only request your email address so that the person you are recommending the page to knows that you wanted them to see it, and that it is not junk mail. We do not capture any email address.

Enter multiple addresses on separate lines or separate them with commas.
Morphine paradoxically prolongs neuropathic pain in rats by amplifying spinal NLRP3 inflammasome activation
(Your Name) has sent you a message from PNAS
(Your Name) thought you would like to see the PNAS web site.
CAPTCHA
This question is for testing whether or not you are a human visitor and to prevent automated spam submissions.
Citation Tools
Morphine prolongs neuropathic pain via NLRP3
Peter M. Grace, Keith A. Strand, Erika L. Galer, Daniel J. Urban, Xiaohui Wang, Michael V. Baratta, Timothy J. Fabisiak, Nathan D. Anderson, Kejun Cheng, Lisa I. Greene, Debra Berkelhammer, Yingning Zhang, Amanda L. Ellis, Hang Hubert Yin, Serge Campeau, Kenner C. Rice, Bryan L. Roth, Steven F. Maier, Linda R. Watkins
Proceedings of the National Academy of Sciences Jun 2016, 113 (24) E3441-E3450; DOI: 10.1073/pnas.1602070113

Citation Manager Formats

  • BibTeX
  • Bookends
  • EasyBib
  • EndNote (tagged)
  • EndNote 8 (xml)
  • Medlars
  • Mendeley
  • Papers
  • RefWorks Tagged
  • Ref Manager
  • RIS
  • Zotero
Request Permissions
Share
Morphine prolongs neuropathic pain via NLRP3
Peter M. Grace, Keith A. Strand, Erika L. Galer, Daniel J. Urban, Xiaohui Wang, Michael V. Baratta, Timothy J. Fabisiak, Nathan D. Anderson, Kejun Cheng, Lisa I. Greene, Debra Berkelhammer, Yingning Zhang, Amanda L. Ellis, Hang Hubert Yin, Serge Campeau, Kenner C. Rice, Bryan L. Roth, Steven F. Maier, Linda R. Watkins
Proceedings of the National Academy of Sciences Jun 2016, 113 (24) E3441-E3450; DOI: 10.1073/pnas.1602070113
del.icio.us logo Digg logo Reddit logo Twitter logo CiteULike logo Facebook logo Google logo Mendeley logo
  • Tweet Widget
  • Facebook Like
  • Mendeley logo Mendeley

Article Classifications

  • Biological Sciences
  • Neuroscience
Proceedings of the National Academy of Sciences: 113 (24)
Table of Contents

Submit

Sign up for Article Alerts

Jump to section

  • Article
    • Abstract
    • Results
    • Discussion
    • Materials and Methods
    • SI Materials and Methods
    • Acknowledgments
    • Footnotes
    • References
  • Figures & SI
  • Info & Metrics
  • PDF

You May Also be Interested in

Water from a faucet fills a glass.
News Feature: How “forever chemicals” might impair the immune system
Researchers are exploring whether these ubiquitous fluorinated molecules might worsen infections or hamper vaccine effectiveness.
Image credit: Shutterstock/Dmitry Naumov.
Reflection of clouds in the still waters of Mono Lake in California.
Inner Workings: Making headway with the mysteries of life’s origins
Recent experiments and simulations are starting to answer some fundamental questions about how life came to be.
Image credit: Shutterstock/Radoslaw Lecyk.
Cave in coastal Kenya with tree growing in the middle.
Journal Club: Small, sharp blades mark shift from Middle to Later Stone Age in coastal Kenya
Archaeologists have long tried to define the transition between the two time periods.
Image credit: Ceri Shipton.
Mouse fibroblast cells. Electron bifurcation reactions keep mammalian cells alive.
Exploring electron bifurcation
Jonathon Yuly, David Beratan, and Peng Zhang investigate how electron bifurcation reactions work.
Listen
Past PodcastsSubscribe
Panda bear hanging in a tree
How horse manure helps giant pandas tolerate cold
A study finds that giant pandas roll in horse manure to increase their cold tolerance.
Image credit: Fuwen Wei.

Similar Articles

Site Logo
Powered by HighWire
  • Submit Manuscript
  • Twitter
  • Facebook
  • RSS Feeds
  • Email Alerts

Articles

  • Current Issue
  • Special Feature Articles – Most Recent
  • List of Issues

PNAS Portals

  • Anthropology
  • Chemistry
  • Classics
  • Front Matter
  • Physics
  • Sustainability Science
  • Teaching Resources

Information

  • Authors
  • Editorial Board
  • Reviewers
  • Subscribers
  • Librarians
  • Press
  • Cozzarelli Prize
  • Site Map
  • PNAS Updates
  • FAQs
  • Accessibility Statement
  • Rights & Permissions
  • About
  • Contact

Feedback    Privacy/Legal

Copyright © 2021 National Academy of Sciences. Online ISSN 1091-6490