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Bed Nucleus of the Stria Terminalis (BNST) neurons containing the serotonin 5HT2c receptor modulate operant alcohol self-administration behavior in mice



The serotonin 5HT2c receptor has been widely implicated in the pathophysiology of alcohol use disorder (AUD), particularly alcohol seeking and the affective consequences of chronic alcohol consumption. However, little is known about the brain sites in which 5HT2c exerts its effects on specific alcohol-related behaviors, especially in females. Here, we investigated the effects of site-specific manipulation of the 5HT2c receptor system in the BNST on operant alcohol self-administration behaviors in adult mice of both sexes, including the acquisition and maintenance of fixed-ratio responding, motivation for alcohol (progressive ratio), and quinine-adulterated responding for alcohol on a fixed-ratio schedule (punished alcohol seeking). Knockdown of 5HT2c in the BNST did not affect the acquisition or maintenance of operant alcohol self-administration, nor did it affect progressive ratio responding for alcohol. This manipulation had only a subtle effect on responding for quinine alcohol selectively in females. On the other hand, chemogenetic inhibition of BNST 5HT2c-containing neurons (BNST5HT2c) increased operant alcohol self-administration behavior in both sexes on day 2, but not day 9, of testing. It also increased operant responding for 1000 μM quinine-adulterated alcohol selectively in males. Importantly, chemogenetic inhibition of BNST5HT2c did not alter operant sucrose responding or motivation for sucrose in either sex. We then performed cell-type specific anterograde tracing, which revealed that BNST5HT2c project to similar regions in males and females, many of which have been previously implicated in AUD. We next used chemogenetics and quantification of the immediate early gene cFos to characterize the functional influence of BNST5HT2c inhibition on vlPAG activity. We show that chemogenetic inhibition of BNST5HT2c reduces vlPAG cFos in both sexes, but that this reduction is more robust in males. Together these findings suggest that BNST5HT2c neurons, and to a small extent the BNST 5HT2c receptor, serve to promote aversive responses to alcohol consumption, potentially through sex-dependent disinhibition of vlPAG neurons.

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Fig. 1: Genetic knockdown of the serotonin 5HT2c receptor in the BNST promotes punished alcohol seeking in females.
Fig. 2: Chemogenetic inhibition of BNST5HT2c promotes punished alcohol seeking in males.
Fig. 3: Chemogenetic inhibition of BNST5HT2c does not influence sucrose seeking.
Fig. 4: BNST5HT2c anterograde tracing in both sexes.
Fig. 5: Chemogenetic inhibition of BNST5HT2c reduces BNST and vlPAG activity in a sex-specific manner.

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Figures for this manuscript were created using The authors would like to acknowledge Drs. Alison Roland and Brianna George for their assistance editing this manuscript.


This work was supported by grants from the National Institutes of Health’s (NIH) National Institute of Alcohol Abuse and Alcoholism (NIAAA) (MEF: K99 AA030628-02; TLK: R01 AA019454-12).

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MEF, CG, and TLK conceived of and designed experiments. MEF, CG, MC, WD, and TS performed behavioral experiments. MEF, MC, and TS performed histology. CG and MEF performed intracranial surgeries. MEF analyzed data, and MEF and TLK wrote manuscript with edits from all authors.

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Correspondence to Thomas L. Kash.

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Flanigan, M.E., Gianessi, C., Castle, M. et al. Bed Nucleus of the Stria Terminalis (BNST) neurons containing the serotonin 5HT2c receptor modulate operant alcohol self-administration behavior in mice. Neuropsychopharmacol. (2023).

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