Article

Deficient endocannabinoid signaling in the central amygdala contributes to alcohol dependence-related anxiety-like behavior and excessive alcohol intake

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Abstract

Negative emotional states that are associated with excessive alcohol intake, particularly anxiety-like states, have been linked to opponent processes in the central nucleus of the amygdala (CeA), affecting stress-related transmitters and monoamines. This study extends these observations to include endocannabinoid signaling in alcohol-dependent animals. Rats and mice were exposed to chronic intermittent alcohol with vapor inhalation or liquid diet to induce dependence. In vivo microdialysis was used to estimate interstitial concentrations of endocannabinoids [N-arachidonoylethanolamine (anandamide; AEA) and 2-arachidonoylglycerol (2-AG)] and amino acids (glutamate and GABA) in rat CeA. Additionally, we evaluated the inhibition of endocannabinoids clearance enzymes [monoacylglycerol lipase (MAGL) and fatty acid amide hydrolase] on anxiety-like behavior and alcohol consumption in alcohol-dependent rats and mice. Results revealed that alcohol dependence produced decreases in baseline 2-AG dialysate levels and increases in baseline levels of glutamate and GABA. Acute alcohol abstinence induced an enhancement of these dependence-induced effects and the levels of 2-AG and GABA were restored upon alcohol re-exposure. Additional studies showed that the increased CeA 2-AG levels induced by restraint stress and alcohol self-administration were blunted in alcohol-dependent rats. Pharmacological studies in rats and mice showed that anxiety-like behavior and alcohol consumption were increased in alcohol-dependent animals, and these behavioral effects were attenuated mainly by MAGL inhibitors [MJN110 (10 and 20 mg/kg) in rats and JZL184 (1 and 3 mg/kg) in mice]. The present results suggest a key role for endocannabinoid signaling in motivational neuroadaptations during alcohol dependence, in which a deficiency in CeA 2-AG signaling in alcohol-dependent animals is linked to stress and excessive alcohol consumption.

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Acknowledgements

We thank Dr. George F. Koob for insightful comments on the manuscript and Mr. Michael Arends for editing the manuscript. We dedicate this manuscript in loving memory to our departed friend, colleague, and mentor, LHP. He was an extraordinary researcher in the field of endocannabinoid signaling, stress, and drug addiction.

Funding and disclosure

This research was supported by the following grants: National Institute on Alcohol Abuse and Alcoholism (AA020404, AA022249, and AA024146 to RMF; AA006420 to RMF and MR; AA017447 and AA015566 to MR; and K99-AA025393 to LAN) and National Institutes of Health (K99/R00DA037344 to JS). This research was also supported by the Pearson Center for Alcoholism and Addiction Research, Instituto de Salud Carlos III (ISCIII) and European Regional Development Funds-European Union (ERDF-EU; Subprograma RETICS Red de Trastornos Adictivos RD12/0028/0001), Ministerio de Economía y Competitividad and ISCIII (PI16/01953 and PI17/02026), Ministerio de Sanidad, Servicios Sociales e Igualdad and Plan Nacional sobre Drogas (PND2017/043), and Consejería de Economía, Innovación y Ciencia, Junta de Andalucía, and ERDF-EU (CTS-433). AS and FJP hold a “Miguel Servet” research contract funded by ISCIII and ERDF-EU (CP14/00173 and CP14/00212, respectively). This is article number 29530 from The Scripps Research Institute.

Author information

Author notes

  1. Deceased: Loren H. Parsons.

Affiliations

  1. Department of Neuroscience, The Scripps Research Institute, La Jolla, CA, USA

    • Antonia Serrano
    • , Francisco J. Pavon
    • , Matthew W. Buczynski
    • , Luis A. Natividad
    • , Ilham Y. Polis
    • , David G. Stouffer
    • , Eric P. Zorrilla
    • , Marisa Roberto
    • , Rémi Martin-Fardon
    •  & Loren H. Parsons
  2. Unidad de Gestión Clínica de Salud Mental, Instituto de Investigación Biomédica de Málaga, Hospital Regional Universitario de Málaga, Málaga, Spain

    • Antonia Serrano
    • , Francisco J. Pavon
    •  & Fernando Rodriguez de Fonseca
  3. Department of Pharmacology & Toxicology, Virginia Commonwealth University, Richmond, VA, USA

    • Joel Schlosburg
  4. Department of Chemical Physiology, Skaggs Institute for Chemical Biology, The Scripps Research Institute, La Jolla, CA, USA

    • Benjamin F. Cravatt

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Conflict of interest

The authors declare that they have no conflict of interest.

Corresponding authors

Correspondence to Antonia Serrano or Rémi Martin-Fardon or Fernando Rodriguez de Fonseca.

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