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Alterations in microbiome composition and metabolic byproducts drive behavioral and transcriptional responses to morphine


Recent evidence has demonstrated that the gut microbiome has marked effects on neuronal function and behavior. Disturbances to microbial populations within the gut have been linked to myriad models of neuropsychiatric disorders. However, the role of the microbiome in substance use disorders remains understudied. Here we show that male mice with their gut microbiome depleted by nonabsorbable antibiotics (Abx) exhibit decreased formation of morphine conditioned place preference across a range of doses (2.5–15 mg/kg), have decreased locomotor sensitization to morphine, and demonstrate marked changes in gene expression within the nucleus accumbens (NAc) in response to high-dose morphine (20 mg/kg × 7 days). Replacement of short-chain fatty acid (SCFA) metabolites, which are reduced by microbiome knockdown, reversed the behavioral and transcriptional effects of microbiome depletion. This identifies SCFA as the crucial mediators of microbiome–brain communication responsible for the effects on morphine reward caused by microbiome knockdown. These studies add important new behavioral, molecular, and mechanistic insight to the role of gut–brain signaling in substance use disorders.

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Fig. 1: Oral Abx alters the microbiome.
Fig. 2: Microbiome knockdown reduces locomotor sensitization and morphine place preference.
Fig. 3: Morphine and Abx alter the NAc transcriptome.
Fig. 4: Microbiome knockdown alters the NAc transcriptional response to morphine.
Fig. 5: Replenishment of SCFAs reverses reward deficit and gene expression changes caused by microbiome depletion.

Data availability

All RNA-sequencing files will be uploaded to the publicly available Gene Expression Omnibus server and will be linked to this publication. All data in this paper will be made available upon reasonable request.


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Morphine sulfate was provided by the NIDA Drug Supply Program. We acknowledge the Microbial Culture & Metabolomics Core of the PennCHOP Microbiome Program for performing targeted metabolomics analyses. Parts of Figs. 1, 2, 3, and 5 were created with with full permission to publsih.

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DDK and RSH designed the experiments. RSH, NLM, TJE, KRM, ATO, and DDK performed experiments. RSH, TJE, KRM, ATO, and DDK analyzed data. RSH and DDK wrote the paper. All authors provided critical edits and feedback of the finalized paper.

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Correspondence to Drew D. Kiraly.

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Hofford, R.S., Mervosh, N.L., Euston, T.J. et al. Alterations in microbiome composition and metabolic byproducts drive behavioral and transcriptional responses to morphine. Neuropsychopharmacol. (2021).

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