Alcohol is a widely consumed substance in our society with great addictive potential (Caslin et al., 2021). Excessive alcohol consumption is a primary risk factor for the development of alcohol use disorders (AUDs), a highly prevalent psychiatric disease with tremendous medical and socioeconomic burdens (Carvalho et al., 2019). Both clinical and animal studies found that there exists individual diversity in alcohol preference (George and Koob, 2017; Gray and MacKillop, 2014; Hisler et al., 2022; Schuckit, 1994). In human beings, individuals who exhibit a lower preference for alcohol (LP) are less likely to engage in excessive drinking or develop alcohol use disorder (AUD) (Gizer et al., 2011; Hendershot et al., 2011), partially due to the negative reactions caused by alcohol accumulation, such as facial flushing, nausea, and increased heart rate (Freeborn et al., 2024; Gross et al., 2015). On the contrary, individuals with higher alcohol preference (HP) obtain greater rewarding effects from alcohol, which increases the risk of AUD (Schuckit, 1994).

Previous studies have reported several brain regions and associated neural circuits involved in alcohol drinking behaviors, providing valuable insights into the effects of alcohol preference on the brain. For example, the mesocorticolimbic circuit has been demonstrated to underpin the reinforcing effects of alcohol consumption (Koob and Volkow, 2016), including ventral tegmental area (VTA) (You et al., 2018), striatum (Clarke and Adermark, 2015; Salinas et al., 2021), and prefrontal cortex (PFC) (Li et al., 2024; Xie et al., 2023). The glutamatergic transmission and gene expression in these regions were altered in high alcohol preferring mouse lines (Bauer et al., 2024; Grecco et al., 2021; Marballi et al., 2016). Additionally, the projection from the central nucleus of the amygdala (CeA) to the bed nucleus of the stria terminalis (BNST) has been confirmed to be involved in the dependence-related escalation of alcohol drinking (de Guglielmo et al., 2019). The orbitofrontal cortex (OFC), medial prefrontal cortex (mPFC), and the dorsal striatum are also implicated in critical neural mechanisms underlying compulsive drinking behavior (Blackwood et al., 2019; Renteria et al., 2018; Timme et al., 2022). However, the differences of neural networks between LP and HP populations remain unknown, which is crucial for understanding the development of AUDs.

In the current study, we aim to provide a detailed comparative analysis of neural network characterizations in mice with different alcohol preference. We parsed the mice into LP group and HP group by two-bottle choice paradigm, and systematically examined the c-Fos profiling in alcohol-related brain areas. Graph theory-based network analysis was used to provide a framework for quantifying the organization of brain networks, identifying key regions (nodes) and their connections (edges) based on measures such as centrality, clustering coefficients, and modularity. As a result, we viewed and distinguished the neural network activity in LP and HP mice holistically and suggested specific brain regions and related neural circuits that could serve as potential targets for pharmacotherapy.