Skip to main content
NCBI home page
Lippincott Open Access logoLink to Lippincott Open Access
. 2026 Feb 26;37(2):117–125. doi: 10.1097/FBP.0000000000000876

Naltrexone treatment improves anxiety- and depression-like behavior in alcohol-exposed mice

Shuangyi Yang 1, Liang Tong 1, Le Zhang 1, Rongzhen Cui 1, Xin Lyu 1, Lin He, Hongyan Liu 1,✉
PMCID: PMC12959587  PMID: 41742765

Abstract

Chronic alcohol exposure threatens central nervous system homeostasis, linking to interconnected dysregulated emotional behavior and robust neuroinflammation. Clinically, individuals with alcohol use disorder (AUD) frequently present comorbid anxiety and depression, yet whether naltrexone, an established therapeutic, can ameliorate AUD neuroinflammation remains unclear. This study investigated naltrexone’s effects on anxiety- and depressive-like behaviors and neuroinflammatory responses in chronically alcohol-exposed mice. Modified ‘Drinking-in-the-Dark’ protocol for 4 weeks, which induced alcohol-exposed mice, followed by subcutaneous naltrexone (1 mg/kg/day) or equal-volume saline for 14 days. Open field test, elevated plus maze test, and tail suspension test are used to detect anxiety- and depressive-like behaviors – alcohol exposure-induced anxiety- and depressive-like behaviors, which naltrexone reversed. ELISA and immunofluorescence revealed alcohol-elevated pro-inflammatory cytokines (tumor necrosis factor-α, interleukin-6, and interleukin-1β) in the basolateral amygdala (BLA), prefrontal cortex, and hippocampus, and promoted BLA microglia proliferation. Naltrexone attenuated these neuroinflammatory changes. These findings highlight naltrexone’s dual-action potential in addressing behavioral and neuroinflammatory consequences of chronic alcohol exposure, providing experimental evidence for its use in AUD, particularly with comorbid anxiety and depression.

Keywords: alcohol, anxiety, depression, naltrexone

Introduction

Ethanol, a widely consumed addictive substance, poses serious global health risks due to misuse. Chronic alcohol use contributes to neurological disorders, such as anxiety, depression, and cognitive deficits, with neuroinflammation playing a key role in these behavioral impairments (Lowe et al., 2020). Targeting neuroinflammatory pathways may therefore offer therapeutic potential for alleviating alcohol-related mood symptoms (Li et al., 2025).

Studies have shown that alcohol exposure induces pro-inflammatory cytokines [e.g. tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6), and interleukin-1β (IL-1β)] (Baxter-Potter et al., 2017) and activates the NOD-like receptor pyrin domain-containing 3 inflammasome, modulating amygdalar synaptic function and anxiety-like behavior (Munshi et al., 2023). Microglia – innate immune cells that release inflammatory factors, alter the immune microenvironment (Wang et al., 2023), and drive abnormal synaptic pruning and cognitive impairment with long-term exposure (Aranda et al., 2021; Soares et al., 2025). Alcohol exposure can activate microglia, and activated glial cells, by producing inflammatory cytokines, serve as an important factor in inducing neuroinflammation (Lowe et al., 2020). Microglia are the innate immune cells of the central nervous system (CNS), and they actively respond to challenges while maintaining brain homeostasis (Wei et al., 2022). Upon stimulation, microglia proliferate and become activated, releasing large amounts of inflammatory factors and altering the immune microenvironment (Wang et al., 2023). Long-term alcohol exposure can trigger microglial-mediated synaptic pruning, leading to cognitive impairments (Aranda et al., 2021; Soares et al., 2025).

Naltrexone, as one of the FDA-approved medications for alcohol use disorder (AUD), can address this inflammatory process (Leung et al., 2022). Naltrexone is an opioid antagonist that can enhance alcohol-induced sedation and other negative alcohol effects while reducing the pleasurable effects of alcohol and alcohol cravings in AUD patients (Krystal et al., 2001; Helstrom et al., 2016). It can reduce neurodegeneration and inflammation (Gatta et al., 2022), exerting neuroprotective effects. Naltrexone can reduce traumatic brain injury-mediated neurodegeneration and inflammation in both wild-type (WT) and mu opioid receptor (MOR) knockout mice (Wang et al., 2021). Therefore, naltrexone may also exert anti-neuroinflammatory effects under alcohol exposure.

While naltrexone has shown effects on alcohol-related behaviors and neuroprotection, its anti-inflammatory role in alcohol-exposed contexts remains unclear. Neuroinflammation serves as a critical shared pathological core for various neurological symptoms. Investigating its anti-inflammatory role in neuroinflammation may help address the limitations of current interventions and provide an efficient translational pathway for repurposing existing drugs in the treatment of related diseases. This study hypothesized that naltrexone can inhibit the excessive proliferation of microglia and reduce neuroinflammation in alcohol-exposed mice, as well as anxiety- and depression-like behaviors. To test this hypothesis, we assessed the effects of naltrexone on anxiety- and depression-like behaviors in chronically alcohol-exposed mice. Subsequently, we investigated the ability of naltrexone to inhibit the excessive proliferation of microglia and inflammatory responses induced by alcohol exposure.

Methods

Subjects

WT C57BL/6J male mice (6–8 weeks) were provided by the Department of Laboratory Animals of Kunming Medical University. The use of animals was approved by the Animal Research Committee of Kunming Medical University (Approval Number: KMMU2024117). All animal experiments were conducted in accordance with the National Institutes of Health (NIH) guidelines for the care and use of laboratory animals. All mice were housed in the animal facility under a 12-hour light/dark cycle at a temperature of 25 ± 2°C and 55% humidity, with free access to standard diet and water. Thirty-six mice were randomly divided into three groups: control group (n = 12), alcohol + saline group (n = 12), and alcohol + naltrexone group (n = 12).

We implemented a modified ‘Drinking-in-the-Dark’ protocol to model binge-like alcohol consumption over extended periods (Belmer et al., 2022). In the alcohol-related group (alcohol + saline group, alcohol + naltrexone group), mice had access to 25% (v/v) alcohol daily (8 PM–8 AM) for 4 weeks. Filtered water was provided adlibitum at all other times. Control group filtered water was provided ad libitum. It aimed to induce neuroinflammation and emotional-behavioral abnormalities triggered by chronic alcohol exposure, and 12-h continuous exposure was adopted to better mimic the long-term consumption patterns of clinical alcohol-dependent individuals, thus ensuring the pathological relevance of the model. A stepwise reduction in sucrose concentration (5% → 2% → 0%) was implemented to eliminate the interference of sucrose-induced taste reward, allowing the mice to gradually adapt to alcohol consumption itself and thereby more authentically reflect the drinking behavior under a state of chronic alcohol exposure.

To establish mouse models of three groups, following the final alcohol exposure, the alcohol + naltrexone group received subcutaneous injections of naltrexone at a dose of 1 mg/kg/day for 14 consecutive days (Zhou et al., 2018). Meanwhile, the alcohol + saline group was administered an equal volume of saline via the same subcutaneous route (Fig. 1). To quantify alcohol consumption, the bottles were weighed on a daily basis; meanwhile, alcohol intake was continuously monitored throughout the modeling period, with the aim of verifying the absence of significant intergroup differences between the two alcohol-exposed cohorts.

Fig. 1.

Fig. 1

Open in a new tab

Flowchart of animal model establishment. In terms of modeling time, from day 1 to day 2, alcohol + naltrexone group and alcohol + saline group were given 10% alcohol + 5% sucrose, control group was given 5% sucrose. From day 3 to day 4, alcohol + naltrexone group and alcohol + saline group were given 15% alcohol + 5% sucrose, and control group was given 5% sucrose. From day 5 to day 7, alcohol + naltrexone group and alcohol + saline group were given 25% alcohol + 2% sucrose, control group was given 2% sucrose. From day 8 to day 28, alcohol + naltrexone group and alcohol + saline group were given 20% alcohol, and control group was given pure water. Monitor the daily alcohol consumption of mice to ensure that there is no difference in their alcohol consumption. EPM, elevated plus maze; OFT, open field test; TST, tail suspension test

Behavioral testing

Behavioral testing was conducted 24 h after the last naltrexone treatment. Behaviors were videorecorded using the Smart 3.0 system (Smart Automobile Co., Ltd., Ningbo, China) for automated post-hoc analysis.

Open field test

The open field test (OFT) is an effective initial screening method for anxiety-like behavior (Kraeuter et al., 2019). In this test, mice are individually placed into a circular arena with a diameter of 30 cm, and the experiment lasts for 5 min. The number of squares crossed by the mice with all four paws is recorded as an indicator of locomotor activity. Additionally, an increase in the time spent in the center of the arena, as well as a decrease in rearing and grooming behaviors (such as fur licking), is considered indicative of anxiolytic-like effects. The floor of the open field is cleaned with 10% ethanol between tests.

Elevated plus maze test

The elevated plus maze (EPM) test is one of the most commonly used methods to evaluate the anxiolytic effects of drugs (Dawson and Tricklebank, 1995). The maze consists of a central platform (6 × 6 cm) and two open arms (30 × 6 cm) that are perpendicular to two closed arms (30 × 6 × 16 cm). The open arms have a 1-cm high plexiglass rim to prevent the mice from falling. The entire apparatus is elevated 50 cm above the ground. During the experiment, mice are individually placed in the central platform, facing the open and closed arms, and their spontaneous behavior is recorded for 5 min. The percentage of total entries into the open arms and the total time spent in the open arms are measured. An increase in the number of entries into the open arms and the time spent there is considered an indicator of anxiolytic behavior. The floor of the EPM is cleaned with 10% ethanol between tests.

Tail suspension test

In this test, the distal end of the mouse’s tail is fixed on a horizontal bar that is 30 cm above the ground, with its head facing downward, maintaining an inverted position (Can et al., 2012). The immobility time is then recorded, with a total recording duration of 5 min. The results primarily reflect the depressive-like behavior exhibited by the mice.

ELISA

The basolateral amygdala (BLA), prefrontal cortex (PFC), and hippocampus (HIP) were dissected from C57BL/6J mouse brains using a coronal sectioning approach (Cao et al., 2017). Following tissue collection, these regions were lysed, and sample concentrations were measured with a bicinchoninic acid assay kit (GBCBIO, Guangzhou, China). Absorbance optical density was read at 450 nm using a BioTek microplate reader (USA).

IL-6, TNF-α, and IL-1β levels in the BLA, PFC, and HIP were quantified using ELISA kits (Proteintech, Wuhan, China) according to the manufacturer’s protocol. Sample sizes ranged from n = 6–8 per group.

Immunofluorescence staining

Following behavioral testing, mice were deeply anesthetized with tribromoethanol (300 mg/kg, intraperitoneal) and transcardially perfused with saline, followed by 4% paraformaldehyde (PFA). Brains were postfixed in 4% PFA for 24 h, cryoprotected in 30% sucrose for 24 h, embedded in optimal cutting temperature compound, and sectioned (40 μm).

Sections were blocked (5% BSA + 0.5% Triton X-100 in PBS, 1 h, room temperature) and incubated with anti-ionized calcium-binding adapter molecule 1 (IBA1) primary antibody (1 : 500, Proteintech, China, 4°C overnight), followed by Alexa Fluor 488-conjugated secondary antibody (Thermo Fisher Scientific, Waltham, USA, 1 h, room temperature). Nuclei were counterstained with 4',6-diamidino-2-phenylindole (Vector Laboratories, Newark, USA), and images were acquired using a confocal microscope (FV3000, Olympus, Tokyo, Japan). Fluorescence signals in the HIP were quantified using Image-Pro Plus 5.0.

Statistical analysis

The t-test was selected to analyze the data of the two groups, and one-way ANOVA was used for comparisons of multiple groups – Bonferroni multiple comparisons post-hoc test. All statistical analyses of experiments were performed by SPSS24.0. Significance was defined as P < 0.05.

Results

Naltrexone treatment alleviates anxiety-like behavior in chronically alcohol-exposed mice

Behavioral tests were conducted to evaluate the efficacy of naltrexone in mitigating anxiety-like behaviors in mice subjected to chronic alcohol exposure. In the EPM test, mice in the alcohol + saline group exhibited significantly reduced time spent in the open arms and fewer entries into the open arms compared with the control group (Fig. 2a–c). OFT revealed that the alcohol + saline group had decreased residence time and fewer entries in the central area relative to the control group (Fig. 2d–f), indicating the induction of anxiety-like behavior by chronic alcohol exposure.

Fig. 2.

Fig. 2

Open in a new tab

Naltrexone treatment improves chronic alcohol exposure-induced anxiety and depression-like behaviors in mice. (a) Trajectory heatmaps of the mouse elevated cross maze representative heatmap of each group; (b and c) Analysis of the number of times the mice entries into the open arm (F = 9.70, DF = 17, P = 0.0020) and the time in the open arm (F = 10.63, DF = 17, P < 0.01; n = 12 each group); (d) trajectory heat maps of the mouse OFT representative heatmap of each group; (e–g) analyzing the entries into the central area (F = 5.55, DF = 17, P < 0.01), the time in the central area (F = 10.02, DF = 17, P < 0.01), and the total distance (F = 19.87, DF = 17, P < 0.001; n = 12 each group); (h) analysis of the immobility of the mice in the TST time (F = 44.95, DF = 17, P < 0.001; n = 12 each group).* P < 0.05, ** P < 0.01, ***P < 0.001. OFT, open field test; TST, tail suspension test.

Notably, alcohol + naltrexone group exhibited significantly increased time spent in the open arms and entries into the open arms compared with alcohol + saline group (Fig. 2a–c). Alcohol + naltrexone group had increased residence time and entries in the central area compared with alcohol + saline group (Fig. 2d–f). These anxiety-related behavioral deficits were markedly ameliorated following two weeks of naltrexone treatment.

Naltrexone treatment alleviates depressive-like behavior in chronically alcohol-exposed mice

Behavioral tests were used to explore the role of naltrexone in ameliorating depression-like behavior in chronic alcohol-exposed mice. OFT test, the total distance moved by the mice in the alcohol + saline group was lower than that of the control group (Fig. 2d and g); tail suspension test (TST) experiment, the duration of immobilization of the mice in the alcohol + saline group was higher than that of the control group (Fig. 2h). The mice showed depressive-like behavior. And the above behaviors were improved after 2 weeks of naltrexone treatment.

Naltrexone treatment alleviates the inflammatory response in the basolateral amygdala region of chronically alcohol-exposed mice

ELISA was used to detect the inflammatory response in the BLA region. The levels of TNF-α, IL-6, and IL-1β in mice in the alcohol + saline group were higher than those in the control group (Fig. 3a–c). TNF-α, IL-6, and IL-1β were improved after 2 weeks of naltrexone treatment.

Fig. 3.

Fig. 3

Open in a new tab

Naltrexone treatment ameliorates the inflammatory response in the BLA region of alcohol-exposed mice. (a–c) Levels of TNF-α (F = 41.37, DF = 17, P < 0.001), IL-6 (F = 43.71, DF = 17, P < 0.001), and IL-1β (F = 23.71, DF = 17, P < 0.001) in the BLA were measured using ELISA kits (n = 8 each group). *P < 0.05, **P < 0.01, ***P < 0.001. BLA, basolateral amygdala; TNF, tumor necrosis factor.

Naltrexone treatment alleviates the inflammatory response in the prefrontal cortex region of chronically alcohol-exposed mice

ELISA was used to detect the inflammatory response in the PFC region. The levels of TNF-α, IL-6, and IL-1β in mice in the alcohol + saline group were higher than those in the control group (Fig. 4a–c). Naltrexone effectively inhibited the increase in TNF-α and IL-6 levels, but there was no change in IL-1β levels.

Fig. 4.

Fig. 4

Open in a new tab

Naltrexone treatment ameliorates the inflammatory response in the PFC region of alcohol-exposed mice. (a–c) Levels of TNF-α (F = 37.65, DF = 17, P < 0.001), IL-6 (F = 16.16, DF = 17, P < 0.001), and IL-1β (F = 15.11, DF = 17, P < 0.001) in the PFC were measured using ELISA kits (n = 8 each group). *P < 0.05, **P < 0.01, ***P < 0.001. PFC, prefrontal cortex.

Naltrexone treatment alleviates the inflammatory response in the hippocampus region of chronically alcohol-exposed mice

ELISA was used to detect the inflammatory response in the HIP region. The levels of TNF-α, IL-6, and IL-1β in mice in the alcohol + saline group were higher than those in the control group (Fig. 5a–c). Naltrexone effectively inhibited the increase in TNF-α and IL-6 levels, but there was no change in IL-1β levels.

Fig. 5.

Fig. 5

Open in a new tab

Naltrexone treatment ameliorates the inflammatory response in the HIP region of alcohol-exposed mice. (a–c) Levels of TNF-α (F = 52.69, DF = 17, P < 0.001), IL-6 (F = 6.89, DF = 17, P < 0.001), and IL-1β (F = 37.34, DF = 17, P < 0.001) in the HIP were measured using ELISA kits (n = 8 each group). *P < 0.05, **P < 0.01, ***P < 0.001. HIP, hippocampus; TNF, tumor necrosis factor.

Naltrexone treatment alleviates microgliocytosis in the basolateral amygdala region of chronically alcohol-exposed mice

IBA1 was used to label microglia, and immunofluorescence assays were performed to detect changes in microglial numbers in the BLA region. The number of IBA1-positive microglia was significantly higher in the alcohol + saline group than in the control group (Fig. 6a and b). This alteration was reversed following a 2-week course of naltrexone treatment. In addition, the count of IBA1-negative cells was also elevated.

Fig. 6.

Fig. 6

Open in a new tab

Naltrexone treatment ameliorates microglial activation in alcohol-exposed mice. (a) Immunofluorescence images of microglia labeled with IBA1 (green) in the BLA region (DAPI, blue, for nuclear counterstaining); each image is a representative result from n = 4 independent mice, with images in each treatment group derived from brain tissues of different mice and corresponding coronal sections of the same brain region to ensure intergroup comparability (n = 4 each group). (b) Quantification of microglial cell numbers in the BLA region (F = 10.53, DF = 11, P < 0.01; n = 4 each group) *P < 0.05, **P < 0.01, ***P < 0.001. BLA, basolateral amygdala; DAPI, 4',6-diamidino-2-phenylindole.

Discussion

Chronic alcohol exposure led to anxiety- and depressive-like behaviors, as evidenced by reduced exploration in the EPM and OFT, as well as increased immobility in the TST. These behavioral alterations are consistent with the well documented effects of chronic alcohol use on emotional dysregulation in both preclinical and clinical settings. The findings presented in this study highlight the multifaceted therapeutic potential of naltrexone in addressing the behavioral and neuroinflammatory consequences of chronic alcohol exposure in mice. The results demonstrate that naltrexone not only alleviates anxiety- and depressive-like behaviors but also mitigates the associated inflammatory response in three key brain regions: the BLA, PFC, and HIP. These brain regions are well established as critical hubs for regulating anxiety and depression, which aligns with prior research highlighting their role in emotional dysregulation linked to alcohol use (Dixon et al., 2017; Wang and Sun, 2021; Johnson and Li, 2022). These behavioral alterations are consistent with the known role of naltrexone as an opioid receptor antagonist, which can modulate the dysregulated reward and stress systems associated with chronic alcohol use (Chamorro et al., 2012; Schacht et al., 2017).

In the CNS, IBA1 is predominantly expressed in microglia (both resting and activated states), whereas being absent in neurons, astrocytes, and oligodendrocytes. When microglia become activated IBA1, expression increases. IBA1 is a standard marker in neuroimmunology studies, with extensive validation in both animal models and human postmortem tissue. Microgliocytosis, characterized by an increased number of IBA1-positive microglia in the BLA region, was observed in alcohol-exposed mice. This microglial activation was accompanied by elevated levels of pro-inflammatory cytokines (TNF-α, IL-6, and IL-1β) (Zheng et al., 2021), indicating a robust inflammatory response (Mu et al., 2022). Naltrexone treatment reduced both microgliocytosis and cytokine levels, suggesting that its therapeutic effects may be partially mediated by the attenuation of neuroinflammation (Munshi et al., 2020; Jiang et al., 2022). This holds true given the growing body of evidence linking neuroinflammation to the pathophysiology of anxiety, depression, and AUD. (Bu et al., 2016; Li et al., 2022). We hypothesize that chronic alcohol exposure may induce apoptosis/loss of cells (neurons or glial cells) in the BLA via mechanisms such as neuroinflammation and oxidative stress, leading to a decrease in nuclear density in both the control group and the alcohol + normal saline group. Interestingly, we also observed an elevation in the number of IBA1-negative cells in the alcohol + naltrexone group. Given that microglial overactivation can trigger the release of pro-inflammatory cytokines, which in turn induce neuronal apoptosis and other forms of cell death (Sha et al., 2025; Qian et al., 2026), naltrexone may exert neuroprotective effects precisely by inhibiting the secretion of these inflammatory factors. This finding is consistent with our observation of reduced cytokine levels following naltrexone treatment.

The findings underscore the potential of naltrexone as a dual-action therapeutic agent that addresses both the behavioral and neuroinflammatory aspects of chronic alcohol exposure. Specifically, given that microglia express MOR (Mali and Novotny, 2022), naltrexone, as an MOR antagonist, may directly bind to microglial MOR to inhibit their abnormal proliferation and the release of pro-inflammatory cytokines (TNF-α, IL-6, and IL-1β). This direct regulation of microglial activity via MOR antagonism likely synergizes with other non-MOR pathways to mitigate alcohol-induced neuroinflammation, which in turn contributes to the amelioration of anxiety- and depressive-like behaviors. This mechanism is supported by Mali and Novotny (Green et al., 2022; Mali and Novotny, 2022), who demonstrated that targeting microglial MOR can effectively modulate neuroinflammatory responses in CNS disorders.

However, several questions remain unanswered. For instance, the precise mechanisms by which naltrexone modulates microglial activation and cytokine release warrant further investigation. Additionally, the long-term effects of naltrexone on neuroinflammation and behavior, as well as its efficacy in different stages of AUC, should be explored. Future studies could also investigate whether the lack of motion in the behavioural tests was due to anxiety- and depression-like effects. In future work, limited dose–response investigations will be conducted to evaluate naltrexone’s neuroinflammatory benefits, and in-vitro cell experiments will be performed to elucidate its specific anti-inflammatory mechanisms. It should be noted that naltrexone did not fully ameliorate all alcohol-related changes.

Conclusion

In summary, this study provides valuable insights into the therapeutic potential of naltrexone in alleviating the behavioral and neuroinflammatory consequences of chronic alcohol exposure. By addressing both the emotional and neurobiological sequelae of alcohol use, naltrexone may offer a promising treatment strategy for AUD who comorbid anxiety and depression, especially in individuals with neuroinflammation. Further research is needed to fully elucidate the mechanisms underlying these effects and to optimize the clinical application of naltrexone in this context.

Acknowledgements

This work was supported by the Scientific Research Fund project of the Education Department of Yunnan Province (2024J0314).

H.L. initiated and designed the project; L.H. and H.L. provided advice and drafted the manuscript; L.T. and L.Z. were responsible for animal modeling and experimental validation; and R.C. and X.L. reviewed relevant literature.

The datasets generated or analyzed during this study areavailable from the corresponding author on reasonable request.

Conflicts of interest

There are no conflicts of interest.

Footnotes

*

Lin He contributed equally to the writing of this article.

References

  1. Aranda J, Fernández-Arjona MDM, Alén F, Rivera P, Rubio L, Smith-Fernández I, et al. (2021). Sudden cessation of fluoxetine before alcohol drinking reinstatement alters microglial morphology and tlr4/inflammatory neuroadaptation in the rat brain. Brain Struct Funct 226:2243–2264. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Baxter-Potter LN, Henricks AM, Berger AL, Bieniasz KV, Lugo JM, Mclaughlin RJ. (2017). Alcohol vapor exposure differentially impacts mesocorticolimbic cytokine expression in a sex-, region-, and duration-specific manner. Neuroscience 346:238–246. [DOI] [PubMed] [Google Scholar]
  3. Belmer A, Depoortere R, Beecher K, Newman-Tancredi A, Bartlett SE. (2022). Neural serotonergic circuits for controlling long-term voluntary alcohol consumption in mice. Mol Psychiatry 27:4599–4610. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Bu W, Ren H, Deng Y, Del Mar N, Guley NM, Moore BM, et al. (2016). Mild traumatic brain injury produces neuron loss that can be rescued by modulating microglial activation using a cb2 receptor inverse agonist. Front Neurosci 10:449. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Can A, Dao DT, Terrillion CE, Piantadosi SC, Bhat S, Gould TD. (2012). The tail suspension test. J Vis Exp 59:e3769. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Cao D, Ma X, Zhang WJ, Xie Z. (2017). Dissection and coronal slice preparation of developing mouse pituitary gland. J Vis Exp 129:56356. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Chamorro AJ, Marcos M, Miron-Canelo JA, Pastor I, Gonzalez-Sarmiento R, Laso FJ. (2012). Association of micro-opioid receptor (oprm1) gene polymorphism with response to naltrexone in alcohol dependence: a systematic review and meta-analysis. Addict Biol 17:505–512. [DOI] [PubMed] [Google Scholar]
  8. Dawson GR, Tricklebank MD. (1995). Use of the elevated plus maze in the search for novel anxiolytic agents. Trends Pharmacol Sci 16:33–36. [DOI] [PubMed] [Google Scholar]
  9. Dixon ML, Thiruchselvam R, Todd R, Christoff K. (2017). Emotion and the prefrontal cortex: an integrative review. Psychol Bull 143:1033–1081. [DOI] [PubMed] [Google Scholar]
  10. Gatta E, Camussi D, Auta J, Guidotti A, Pandey SC. (2022). Neurosteroids (allopregnanolone) and alcohol use disorder: from mechanisms to potential pharmacotherapy. Pharmacol Ther 240:108299. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Green JM, Sundman MH, Chou Y. (2022). Opioid-induced microglia reactivity modulates opioid reward, analgesia, and behavior. Neurosci Biobehav Rev 135:104544. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Helstrom AW, Blow FC, Slaymaker V, Kranzler HR, Leong S, Oslin D. (2016). Reductions in alcohol craving following naltrexone treatment for heavy drinking. Alcohol Alcohol 51:562–566. [DOI] [PubMed] [Google Scholar]
  13. Jiang L, Hao J, Yang XL, Zhu JX, Wang Y, Huang YL, et al. (2022). Basolateral amygdala reactive microglia may contribute to synaptic impairment and depressive-like behavior in mice with bone cancer pain. Neurochem Res 47:3454–3463. [DOI] [PubMed] [Google Scholar]
  14. Johnson J, Li Z. (2022). Fuel for emotion: how mitophagy in the BLA can mediate increased anxiety in stressful social situations. Autophagy 18:465–466. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Kraeuter AK, Guest PC, Sarnyai Z. (2019). The open field test for measuring locomotor activity and anxiety-like behavior. Methods Mol Biol 1916:99–103. [DOI] [PubMed] [Google Scholar]
  16. Krystal JH, Cramer JA, Krol WF, Kirk GF, Rosenheck RA; Veterans Affairs Naltrexone Cooperative Study 425 Group (2001). Naltrexone in the treatment of alcohol dependence. N Engl J Med 345:1734–1739. [DOI] [PubMed] [Google Scholar]
  17. Leung JG, Narayanan PP, Markota M, Miller NE, Philbrick KL, Burton MC, Kirchoff RW. (2022). Assessing naltrexone prescribing and barriers to initiation for alcohol use disorder: a multidisciplinary, multisite survey. Front Psychiatry 13:856938. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Li C, Wu H, Sen Ta Na H, Wang L, Zhong C, Deng B, et al. (2022). Neuronal-microglial liver x receptor beta activating decrease neuroinflammation and chronic stress-induced depression-related behavior in mice. Brain Res 1797:148112. [DOI] [PubMed] [Google Scholar]
  19. Li X, Ni Z, Shi W, Zhao K, Zhang Y, Liu L, et al. (2025). Nitrate ameliorates alcohol-induced cognitive impairment via oral microbiota. J Neuroinflammation 22:106. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Lowe PP, Morel C, Ambade A, Iracheta-Vellve A, Kwiatkowski E, Satishchandran A, et al. (2020). Chronic alcohol-induced neuroinflammation involves ccr2/5-dependent peripheral macrophage infiltration and microglia alterations. J Neuroinflammation 17:296. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Mali AS, Novotny J. (2022). Opioid receptor activation suppresses the neuroinflammatory response by promoting microglial m2 polarization. Mol Cell Neurosci 121:103744. [DOI] [PubMed] [Google Scholar]
  22. Mu JL, Liu XD, Dong YH, Fang YY, Qiu SD, Zhang F, Liu KX. (2022). Peripheral interleukin-6-associated microglial quin elevation in basolateral amygdala contributed to cognitive dysfunction in a mouse model of postoperative delirium. Front Med (Lausanne) 9:998397. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Munshi S, Loh MK, Ferrara N, Dejoseph MR, Ritger A, Padival M, et al. (2020). Repeated stress induces a pro-inflammatory state, increases amygdala neuronal and microglial activation, and causes anxiety in adult male rats. Brain Behav Immun 84:180–199. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Munshi S, Albrechet-Souza L, Dos-Santos RC, Stelly CE, Secci ME, Gilpin NW, Tasker JG. (2023). Acute ethanol modulates synaptic inhibition in the basolateral amygdala via rapid nlrp3 inflammasome activation and regulates anxiety-like behavior in rats. J Neurosci 43:7902–7912. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Qian Y, Yu H, Dong J, Liu J, Han J, Zheng T, et al. (2026). Microglial tia1-mediated stress granules promote neuroinflammation and aggravate neuron loss in mice after ischemic stroke by inhibiting igf2 signaling. Theranostics 16:2627–2648. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Schacht JP, Randall PK, Latham PK, Voronin KE, Book SW, Myrick H, Anton RF. (2017). Predictors of naltrexone response in a randomized trial: reward-related brain activation, oprm1 genotype, and smoking status. Neuropsychopharmacology 42:2640–2653. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Sha Z, Dong S, Nie M, Liu T, Wu C, Lv C, et al. (2025). Genetic deletion of g protein-coupled receptor 56 aggravates traumatic brain injury through the microglial ccl3/4/5 upregulation targeted to ccr5. Cell Death Dis 16:175. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Soares AR, Garcia-Rivas V, Fai C, Thomas M, Zheng X, Picciotto MR, Mineur YS. (2025). Sex differences in the microglial response to stress and chronic alcohol exposure in mice. Biol Sex Differ 16:19. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Wang Y, Sun QQ. (2021). A long-range, recurrent neuronal network linking the emotion regions with the somatic motor cortex. Cell Rep 36:109733. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Wang YS, Hung TW, Bae EK, Wu KJ, Hsieh W, Yu SJ. (2021). Naltrexone is neuroprotective against traumatic brain injury in mu opioid receptor knockout mice. CNS Neurosci Ther 27:831–841. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Wang S, Xu C, Liu H, Wei W, Zhou X, Qian H, et al. (2023). Connecting the gut microbiota and neurodegenerative diseases: the role of bile acids. Mol Neurobiol 60:4618–4640. [DOI] [PubMed] [Google Scholar]
  32. Wei W, Wang S, Xu C, Zhou X, Lian X, He L, Li K. (2022). Gut microbiota, pathogenic proteins and neurodegenerative diseases. Front Microbiol 13:959856. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Zheng ZH, Tu JL, Li XH, Hua Q, Liu WZ, Liu Y, et al. (2021). Neuroinflammation induces anxiety- and depressive-like behavior by modulating neuronal plasticity in the basolateral amygdala. Brain Behav Immun 91:505–518. [DOI] [PubMed] [Google Scholar]
  34. Zhou Y, Crowley R, Prisinzano T, Kreek MJ. (2018). Effects of mesyl salvinorin b alone and in combination with naltrexone on alcohol deprivation effect in male and female mice. Neurosci Lett 673:19–23. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Behavioural Pharmacology are provided here courtesy of Wolters Kluwer Health

RESOURCES