Spontaneous and Naloxone-Precipitated Withdrawal Behaviors From Chronic Opiates are Accompanied by Changes in N-Oleoylglycine and N-Oleoylalanine Levels in the Brain and Ameliorated by Treatment With These Mediators

Samantha M. Ayoub, Fabiana Piscitelli, Cristoforo Silvestri, Cheryl L. Limebeer, Erin M. Rock, Reem Smoum, Mathew Farag, Hannah de Almeida, Megan T. Sullivan, Sébastien Lacroix, Besma Boubertakh, Nayudu Nallabelli, Aron H. Lichtman, Francesco Leri, Raphael Mechoulam, Vincenzo Di Marzo, Linda A. Parker*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

7 Scopus citations


Rationale: The endocannabinoidome mediators, N-Oleoylglycine (OlGly) and N-Oleoylalanine (OlAla), have been shown to reduce acute naloxone-precipitated morphine withdrawal affective and somatic responses. Objectives: To determine the role and mechanism of action of OlGly and OlAla in withdrawal responses from chronic exposure to opiates in male Sprague-Dawley rats. Methods: Opiate withdrawal was produced: 1) spontaneously 24 h following chronic exposure to escalating doses of morphine over 14 days (Experiments 1 and 2) and steady-state exposure to heroin by minipumps for 12 days (Experiment 3), 2) by naloxone injection during steady-state heroin exposure (Experiment 4), 3) by naloxone injection during operant heroin self-administration (Experiment 5). Results: In Experiment 1, spontaneous morphine withdrawal produced somatic withdrawal reactions. The behavioral withdrawal reactions were accompanied by suppressed endogenous levels of OlGly in the nucleus accumbens, amygdala, and prefrontal cortex, N-Arachidonylglycerol and OlAla in the amygdala, 2-arachidonoylglycerol in the nucleus accumbens, amygdala and interoceptive insular cortex, and by changes in colonic microbiota composition. In Experiment 2, treatment with OlAla, but not OlGly, reduced spontaneous morphine withdrawal responses. In Experiment 3, OlAla attenuated spontaneous steady-state heroin withdrawal responses at both 5 and 20 mg/kg; OlGly only reduced withdrawal responses at the higher dose of 20 mg/kg. Experiment 4 demonstrated that naloxone-precipitated heroin withdrawal from steady-state exposure to heroin (7 mg/kg/day for 12 days) is accompanied by tissue-specific changes in brain or gut endocannabinoidome mediator, including OlGly and OlAla, levels and colonic microbiota composition, and that OlAla (5 mg/kg) attenuated behavioural withdrawal reactions, while also reversing some of the changes in brain and gut endocannabinoidome and gut microbiota induced by naloxone. Experiment 5 demonstrated that although OlAla (5 mg/kg) did not interfere with operant heroin self-administration on its own, it blocked naloxone-precipitated elevation of heroin self-administration behavior. Conclusion: These results suggest that OlAla and OlGly are two endogenous mediators whose brain concentrations respond to chronic opiate treatment and withdrawal concomitantly with changes in colon microbiota composition, and that OlAla may be more effective than OlGly in suppressing chronic opiate withdrawal responses.

Original languageAmerican English
Article number706703
JournalFrontiers in Pharmacology
StatePublished - 15 Sep 2021

Bibliographical note

Funding Information:
This research was supported by research funding to 1) LAP from the Natural Sciences and Engineering Research Council (NSERC 03629), the Canadian Institutes of Health Research (CIHR-388239) and PlantExt. 2) VD from the Canada Research Excellence Chair on the Microbiome-Endocannabinoidome

Publisher Copyright:
© Copyright © 2021 Ayoub, Piscitelli, Silvestri, Limebeer, Rock, Smoum, Farag, de Almeida, Sullivan, Lacroix, Boubertakh, Nallabelli, Lichtman, Leri, Mechoulam, Di Marzo and Parker.


  • CB
  • PPARalpha
  • endocannabinoidome
  • heroin self-administration
  • oleoyl alanine
  • oleoyl glycine
  • opiate withdrawal
  • somatic withdrawal
  • CB1


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