{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,1]],"date-time":"2026-07-01T08:49:05Z","timestamp":1782895745030,"version":"3.54.5"},"reference-count":86,"publisher":"Springer Science and Business Media LLC","issue":"1","license":[{"start":{"date-parts":[[2026,4,30]],"date-time":"2026-04-30T00:00:00Z","timestamp":1777507200000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0"},{"start":{"date-parts":[[2026,7,1]],"date-time":"2026-07-01T00:00:00Z","timestamp":1782864000000},"content-version":"vor","delay-in-days":62,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0"}],"funder":[{"DOI":"10.13039\/100000002","name":"National Institutes of Health","doi-asserted-by":"publisher","award":["K00 AA029955"],"award-info":[{"award-number":["K00 AA029955"]}],"id":[{"id":"10.13039\/100000002","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100000002","name":"National Institutes of Health","doi-asserted-by":"publisher","award":["AA012404 ;  AA020926"],"award-info":[{"award-number":["AA012404 ;  AA020926"]}],"id":[{"id":"10.13039\/100000002","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["link.springer.com"],"crossmark-restriction":false},"short-container-title":["Sci Rep"],"abstract":"<jats:title>Abstract<\/jats:title>\n                  <jats:p>\n                    Alcohol consumption triggers neuroinflammation, potentially creating a feed-forward loop that increases drinking. Previous studies have shown that activation of the toll-like receptor 7 (TLR7) leads to escalated drinking. In this study, we aim to identify cell type-specific transcriptomic patterns underlying TLR7-induced neuroinflammation, potentially leading to escalated drinking. Therefore, male C57BL\/6J mice were treated with the selective TLR7 agonist R848 every-other day for 20 days in total. After 10 treatment-free days, half of the cohort underwent two bottle-choice drinking; the other half was sacrificed and brains were collected for single-nucleus RNA-Sequencing (snRNA-Seq) in the medial prefrontal cortex (mPFC) and central amygdala (AMG). The AMG showed a greater number of differentially expressed genes (DEGs), primarily in inhibitory and excitatory neurons. Among glial cells, AMG astrocytes exhibited the greatest number of DEGs, which were involved in blood-brain barrier (BBB) regulation (e.g.,\n                    <jats:italic>Cldn5<\/jats:italic>\n                    ,\n                    <jats:italic>Mecom<\/jats:italic>\n                    ,\n                    <jats:italic>Nrg1<\/jats:italic>\n                    ), a finding supported by secondary validation using Xenium in situ spatial transcriptomics on AMG-containing sections. BBB-regulatory genes, including those in the Wnt signaling pathway (e.g.,\n                    <jats:italic>Notch3<\/jats:italic>\n                    ,\n                    <jats:italic>Top2a<\/jats:italic>\n                    ,\n                    <jats:italic>Aldoc<\/jats:italic>\n                    ), were altered across multiple cell types in both regions. Together with the alterations observed in neurons, these findings suggest that repeated TLR7 activation induces persistent BBB and neuronal dysregulation, potentially leading to TLR7-induced escalated drinking.\n                  <\/jats:p>","DOI":"10.1038\/s41598-026-50920-5","type":"journal-article","created":{"date-parts":[[2026,4,30]],"date-time":"2026-04-30T12:25:10Z","timestamp":1777551910000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Repeated TLR7 activation induces cell type- and brain region-specific transcriptome changes in male mice"],"prefix":"10.1038","volume":"16","author":[{"given":"Marion M","family":"Friske","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Riccardo","family":"Barchiesi","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Nihal A","family":"Salem","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ruth L","family":"Allard","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Anna C","family":"Dobre","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Nicholas","family":"Rhyan","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Wen","family":"Chen","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"R Dayne","family":"Mayfield","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"297","published-online":{"date-parts":[[2026,4,30]]},"reference":[{"key":"50920_CR1","doi-asserted-by":"publisher","first-page":"922","DOI":"10.1001\/jamapsychiatry.2021.1271","volume":"78","author":"B Han","year":"2021","unstructured":"Han, B., Jones, C. M., Einstein, E. B., Powell, P. A. & Compton, W. M. Use of Medications for Alcohol Use Disorder in the US: Results From the 2019 National Survey on Drug Use and Health. JAMA Psychiatry. 78, 922\u2013924 (2021).","journal-title":"JAMA Psychiatry"},{"key":"50920_CR2","doi-asserted-by":"publisher","first-page":"498","DOI":"10.1111\/acer.14792","volume":"46","author":"I Sohi","year":"2022","unstructured":"Sohi, I. et al. Changes in alcohol use during the COVID-19 pandemic and previous pandemics: A systematic review. Alcohol Clin. Exp. Res. 46, 498\u2013513 (2022).","journal-title":"Alcohol Clin. Exp. Res."},{"key":"50920_CR3","doi-asserted-by":"publisher","first-page":"13","DOI":"10.1016\/B978-0-12-801284-0.00002-6","volume":"118","author":"G Robinson","year":"2014","unstructured":"Robinson, G. et al. Neuroimmune pathways in alcohol consumption: Evidence from behavioral and genetic studies in rodents and humans. Int. Rev. Neurobiol. 118, 13\u201339 (2014).","journal-title":"Int. Rev. Neurobiol."},{"key":"50920_CR4","doi-asserted-by":"publisher","first-page":"2081","DOI":"10.2217\/pgs-2016-0062","volume":"17","author":"A Warden","year":"2016","unstructured":"Warden, A., Erickson, E., Robinson, G., Harris, R. A. & Mayfield, R. D. The neuroimmune transcriptome and alcohol dependence: Potential for targeted therapies. Pharmacogenomics 17, 2081\u20132096 (2016).","journal-title":"Pharmacogenomics"},{"key":"50920_CR5","doi-asserted-by":"publisher","first-page":"376","DOI":"10.1038\/npp.2016.177","volume":"42","author":"J Mayfield","year":"2017","unstructured":"Mayfield, J. & Harris, R. A. The Neuroimmune Basis of Excessive Alcohol Consumption. Neuropsychopharmacology 42, 376 (2017).","journal-title":"Neuropsychopharmacology"},{"key":"50920_CR6","doi-asserted-by":"publisher","first-page":"315","DOI":"10.1016\/B978-0-12-801284-0.00010-5","volume":"118","author":"FT Crews","year":"2014","unstructured":"Crews, F. T. & Vetreno, R. P. Neuroimmune basis of alcoholic brain damage. Int. Rev. Neurobiol. 118, 315\u2013357 (2014).","journal-title":"Int. Rev. Neurobiol."},{"key":"50920_CR7","doi-asserted-by":"publisher","first-page":"939","DOI":"10.1111\/acer.13368","volume":"41","author":"CJ Lawrimore","year":"2017","unstructured":"Lawrimore, C. J., Crews, F. T. & Ethanol TLR3, and TLR4 Agonists Have Unique Innate Immune Responses in Neuron-Like SH-SY5Y and Microglia-Like BV2. Alcohol Clin. Exp. Res. 41, 939\u2013954 (2017).","journal-title":"Alcohol Clin. Exp. Res."},{"key":"50920_CR8","doi-asserted-by":"publisher","first-page":"66","DOI":"10.1016\/j.bbi.2018.12.006","volume":"77","author":"AS Warden","year":"2019","unstructured":"Warden, A. S. et al. Toll-like receptor 3 dynamics in female C57BL\/6J mice: Regulation of alcohol intake. Brain Behav. Immun. 77, 66\u201376 (2019).","journal-title":"Brain Behav. Immun."},{"key":"50920_CR9","doi-asserted-by":"publisher","first-page":"173491","DOI":"10.1016\/j.pbb.2022.173491","volume":"222","author":"EK Grantham","year":"2023","unstructured":"Grantham, E. K., Barchiesi, R., Salem, N. A. & Mayfield, R. D. Neuroimmune pathways as targets to reduce alcohol consumption. Pharmacol. Biochem. Behav. 222, 173491 (2023).","journal-title":"Pharmacol. Biochem. Behav."},{"key":"50920_CR10","doi-asserted-by":"publisher","first-page":"22","DOI":"10.1186\/s12974-017-0799-4","volume":"14","author":"LG Coleman Jr","year":"2017","unstructured":"Coleman, L. G. Jr, Zou, J. & Crews, F. T. Microglial-derived miRNA let-7 and HMGB1 contribute to ethanol-induced neurotoxicity via TLR7. J. Neuroinflammation. 14, 22 (2017).","journal-title":"J. Neuroinflammation"},{"key":"50920_CR11","doi-asserted-by":"publisher","first-page":"2547","DOI":"10.3390\/ijms22052547","volume":"22","author":"L Qin","year":"2021","unstructured":"Qin, L. et al. TRAIL Mediates Neuronal Death in AUD: A Link between Neuroinflammation and Neurodegeneration. Int. J. Mol. Sci. 22, 2547 (2021).","journal-title":"Int. J. Mol. Sci."},{"key":"50920_CR12","doi-asserted-by":"publisher","first-page":"602","DOI":"10.1016\/j.biopsych.2012.09.030","volume":"73","author":"FT Crews","year":"2013","unstructured":"Crews, F. T., Qin, L., Sheedy, D., Vetreno, R. P. & Zou, J. High mobility group box 1\/Toll-like receptor danger signaling increases brain neuroimmune activation in alcohol dependence. Biol. Psychiatry. 73, 602\u2013612 (2013).","journal-title":"Biol. Psychiatry"},{"key":"50920_CR13","doi-asserted-by":"publisher","first-page":"516","DOI":"10.1111\/acer.13316","volume":"41","author":"YA Blednov","year":"2017","unstructured":"Blednov, Y. A. et al. Ethanol consumption in mice lacking CD14, TLR2, TLR4, or MyD88. Alcohol Clin. Exp. Res. 41, 516\u2013530 (2017).","journal-title":"Alcohol Clin. Exp. Res."},{"key":"50920_CR14","doi-asserted-by":"publisher","first-page":"245","DOI":"10.1016\/j.bbi.2014.12.019","volume":"45","author":"F Corrigan","year":"2015","unstructured":"Corrigan, F. et al. Alcohol-induced sedation and synergistic interactions between alcohol and morphine: A key mechanistic role for Toll-like receptors and MyD88-dependent signaling. Brain Behav. Immun. 45, 245\u2013252 (2015).","journal-title":"Brain Behav. Immun."},{"key":"50920_CR15","doi-asserted-by":"publisher","first-page":"e12932","DOI":"10.1111\/adb.12932","volume":"26","author":"YA Blednov","year":"2021","unstructured":"Blednov, Y. A., Da Costa, A., Mayfield, J., Harris, R. A. & Messing, R. O. Deletion of Tlr3 reduces acute tolerance to alcohol and alcohol consumption in the intermittent access procedure in male mice. Addict. Biol. 26, e12932 (2021).","journal-title":"Addict. Biol."},{"key":"50920_CR16","doi-asserted-by":"publisher","first-page":"437","DOI":"10.1016\/j.bbi.2024.03.021","volume":"118","author":"GA Dilly","year":"2024","unstructured":"Dilly, G. A. et al. Knockdown of Tlr3 in dorsal striatum reduces ethanol consumption and acute functional tolerance in male mice. Brain Behav. Immun. 118, 437\u2013448 (2024).","journal-title":"Brain Behav. Immun."},{"key":"50920_CR17","doi-asserted-by":"publisher","first-page":"423","DOI":"10.1016\/j.bbi.2020.07.029","volume":"89","author":"EK Grantham","year":"2020","unstructured":"Grantham, E. K. et al. Role of toll-like receptor 7 (TLR7) in voluntary alcohol consumption. Brain Behav. Immun. 89, 423\u2013432 (2020).","journal-title":"Brain Behav. Immun."},{"key":"50920_CR18","doi-asserted-by":"publisher","first-page":"e13176","DOI":"10.1111\/adb.13176","volume":"27","author":"DF Lovelock","year":"2022","unstructured":"Lovelock, D. F. et al. The Toll-like receptor 7 agonist imiquimod increases ethanol self-administration and induces expression of Toll-like receptor related genes. Addict. Biol. 27, e13176 (2022).","journal-title":"Addict. Biol."},{"key":"50920_CR19","doi-asserted-by":"publisher","first-page":"578","DOI":"10.1038\/s41397-017-0012-2","volume":"18","author":"EK Erickson","year":"2018","unstructured":"Erickson, E. K., Farris, S. P., Blednov, Y. A., Mayfield, R. D. & Harris, R. A. Astrocyte-specific transcriptome responses to chronic ethanol consumption. Pharmacogenomics J. 18, 578\u2013589 (2018).","journal-title":"Pharmacogenomics J."},{"key":"50920_CR20","doi-asserted-by":"publisher","first-page":"10949","DOI":"10.1038\/s41598-019-47454-4","volume":"9","author":"EK Erickson","year":"2019","unstructured":"Erickson, E. K., Blednov, Y. A., Harris, R. A. & Mayfield, R. D. Glial gene networks associated with alcohol dependence. Sci. Rep. 9, 10949 (2019).","journal-title":"Sci. Rep."},{"key":"50920_CR21","doi-asserted-by":"publisher","first-page":"34","DOI":"10.1016\/j.pbb.2018.12.007","volume":"177","author":"EK Erickson","year":"2019","unstructured":"Erickson, E. K., Grantham, E. K., Warden, A. S. & Harris, R. A. Neuroimmune signaling in alcohol use disorder. Pharmacol. Biochem. Behav. 177, 34\u201360 (2019).","journal-title":"Pharmacol. Biochem. Behav."},{"key":"50920_CR22","doi-asserted-by":"publisher","first-page":"89","DOI":"10.1038\/s41398-019-0384-y","volume":"9","author":"M Kapoor","year":"2019","unstructured":"Kapoor, M. et al. Analysis of whole genome-transcriptomic organization in brain to identify genes associated with alcoholism. Transl Psychiatry. 9, 89 (2019).","journal-title":"Transl Psychiatry"},{"key":"50920_CR23","doi-asserted-by":"publisher","first-page":"275","DOI":"10.3390\/brainsci10050275","volume":"10","author":"SP Farris","year":"2020","unstructured":"Farris, S. P. et al. Transcriptome Analysis of Alcohol Drinking in Non-Dependent and Dependent Mice Following Repeated Cycles of Forced Swim Stress Exposure. Brain Sci. 10, 275 (2020).","journal-title":"Brain Sci."},{"key":"50920_CR24","doi-asserted-by":"publisher","first-page":"1760","DOI":"10.1111\/acer.14410","volume":"44","author":"AS Warden","year":"2020","unstructured":"Warden, A. S. et al. Inbred substrain differences influence neuroimmune response and drinking behavior. Alcohol Clin. Exp. Res. 44, 1760\u20131768 (2020).","journal-title":"Alcohol Clin. Exp. Res."},{"key":"50920_CR25","doi-asserted-by":"publisher","first-page":"910","DOI":"10.1016\/j.biopsych.2020.05.011","volume":"88","author":"AS Warden","year":"2020","unstructured":"Warden, A. S. et al. Microglia control escalation of drinking in alcohol-dependent mice: Genomic and synaptic drivers. Biol. Psychiatry. 88, 910\u2013921 (2020).","journal-title":"Biol. Psychiatry"},{"key":"50920_CR26","doi-asserted-by":"publisher","first-page":"760","DOI":"10.1016\/S2215-0366(16)00104-8","volume":"3","author":"GF Koob","year":"2016","unstructured":"Koob, G. F. & Volkow, N. D. Neurobiology of addiction: A neurocircuitry analysis. Lancet Psychiatry. 3, 760\u2013773 (2016).","journal-title":"Lancet Psychiatry"},{"key":"50920_CR27","doi-asserted-by":"publisher","DOI":"10.1038\/s41380-024-02719-x","author":"MM Friske","year":"2024","unstructured":"Friske, M. M. et al. A systematic review and meta-analysis on the transcriptomic signatures in alcohol use disorder. Mol. Psychiatry 2024 September. https:\/\/doi.org\/10.1038\/s41380-024-02719-x (2024).","journal-title":"Mol. Psychiatry 2024 September"},{"key":"50920_CR28","doi-asserted-by":"publisher","first-page":"a039339","DOI":"10.1101\/cshperspect.a039339","volume":"11","author":"M Roberto","year":"2021","unstructured":"Roberto, M., Kirson, D. & Khom, S. The role of the central amygdala in alcohol dependence. Cold Spring Harb Perspect. Med. 11, a039339 (2021).","journal-title":"Cold Spring Harb Perspect. Med."},{"key":"50920_CR29","doi-asserted-by":"publisher","first-page":"926","DOI":"10.1111\/acer.13616","volume":"42","author":"YA Blednov","year":"2018","unstructured":"Blednov, Y. A. et al. Apremilast alters behavioral responses to ethanol in mice: I. reduced consumption and preference. Alcohol Clin. Exp. Res. 42, 926\u2013938 (2018).","journal-title":"Alcohol Clin. Exp. Res."},{"key":"50920_CR30","doi-asserted-by":"publisher","first-page":"1938","DOI":"10.1111\/j.1530-0277.2011.01545.x","volume":"35","author":"LS Hwa","year":"2011","unstructured":"Hwa, L. S. et al. Persistent escalation of alcohol drinking in C57BL\/6J mice with intermittent access to 20% ethanol. Alcohol Clin. Exp. Res. 35, 1938\u20131947 (2011).","journal-title":"Alcohol Clin. Exp. Res."},{"key":"50920_CR31","doi-asserted-by":"publisher","first-page":"4169","DOI":"10.1038\/s41467-019-12054-3","volume":"10","author":"A Bhattacherjee","year":"2019","unstructured":"Bhattacherjee, A. et al. Cell type-specific transcriptional programs in mouse prefrontal cortex during adolescence and addiction. Nat. Commun. 10, 4169 (2019).","journal-title":"Nat. Commun."},{"key":"50920_CR32","doi-asserted-by":"publisher","first-page":"1880","DOI":"10.1038\/s41593-023-01455-9","volume":"26","author":"A Bhattacherjee","year":"2023","unstructured":"Bhattacherjee, A. et al. Spatial transcriptomics reveals the distinct organization of mouse prefrontal cortex and neuronal subtypes regulating chronic pain. Nat. Neurosci. 26, 1880\u20131893 (2023).","journal-title":"Nat. Neurosci."},{"key":"50920_CR33","doi-asserted-by":"publisher","first-page":"2237","DOI":"10.1038\/s41593-023-01469-3","volume":"26","author":"H Hochgerner","year":"2023","unstructured":"Hochgerner, H. et al. Neuronal types in the mouse amygdala and their transcriptional response to fear conditioning. Nat. Neurosci. 26, 2237\u20132249 (2023).","journal-title":"Nat. Neurosci."},{"key":"50920_CR34","doi-asserted-by":"publisher","first-page":"317","DOI":"10.1038\/s41586-023-06812-z","volume":"624","author":"Z Yao","year":"2023","unstructured":"Yao, Z. et al. A high-resolution transcriptomic and spatial atlas of cell types in the whole mouse brain. Nature 624, 317\u2013332 (2023).","journal-title":"Nature"},{"key":"50920_CR35","doi-asserted-by":"publisher","first-page":"289","DOI":"10.1038\/s41398-022-02063-0","volume":"12","author":"GA Dilly","year":"2022","unstructured":"Dilly, G. A., Kittleman, C. W., Kerr, T. M., Messing, R. O. & Mayfield, R. D. Cell-type specific changes in PKC-delta neurons of the central amygdala during alcohol withdrawal. Transl Psychiatry. 12, 289 (2022).","journal-title":"Transl Psychiatry"},{"key":"50920_CR36","doi-asserted-by":"publisher","first-page":"338","DOI":"10.1016\/j.bbi.2019.09.004","volume":"82","author":"KA Michaelis","year":"2019","unstructured":"Michaelis, K. A. et al. Persistent Toll-like receptor 7 stimulation induces behavioral and molecular innate immune tolerance. Brain Behav. Immun. 82, 338\u2013353 (2019).","journal-title":"Brain Behav. Immun."},{"key":"50920_CR37","doi-asserted-by":"publisher","first-page":"100639","DOI":"10.1016\/j.ynstr.2024.100639","volume":"31","author":"RL Allard","year":"2024","unstructured":"Allard, R. L., Mayfield, J., Barchiesi, R., Salem, N. A. & Mayfield, R. D. Toll-like receptor 7: A novel neuroimmune target to reduce excessive alcohol consumption. Neurobiol. Stress. 31, 100639 (2024).","journal-title":"Neurobiol. Stress"},{"key":"50920_CR38","doi-asserted-by":"publisher","first-page":"106361","DOI":"10.1016\/j.nbd.2023.106361","volume":"190","author":"NA Salem","year":"2024","unstructured":"Salem, N. A. et al. Cell-type brain-region specific changes in prefrontal cortex of a mouse model of alcohol dependence. Neurobiol. Dis. 190, 106361 (2024).","journal-title":"Neurobiol. Dis."},{"key":"50920_CR39","doi-asserted-by":"publisher","first-page":"377","DOI":"10.1038\/s41586-023-06816-9","volume":"625","author":"A Cui","year":"2024","unstructured":"Cui, A. et al. Dictionary of immune responses to cytokines at single-cell resolution. Nature 625, 377\u2013384 (2024).","journal-title":"Nature"},{"key":"50920_CR40","doi-asserted-by":"publisher","first-page":"163","DOI":"10.1016\/j.bbi.2022.02.016","volume":"102","author":"C Acioglu","year":"2022","unstructured":"Acioglu, C., Heary, R. F. & Elkabes, S. Roles of neuronal toll-like receptors in neuropathic pain and central nervous system injuries and diseases. Brain Behav. Immun. 102, 163\u2013178 (2022).","journal-title":"Brain Behav. Immun."},{"key":"50920_CR41","doi-asserted-by":"publisher","unstructured":"Warden, A. S. et al. Integrative genomics approach identifies glial transcriptomic dysregulation and risk in the cortex of individuals with Alcohol Use Disorder. Biol. Psychiatry 2025 Febr. 28 https:\/\/doi.org\/10.1016\/j.biopsych.2025.02.895 (2025).","DOI":"10.1016\/j.biopsych.2025.02.895"},{"key":"50920_CR42","doi-asserted-by":"publisher","first-page":"1634","DOI":"10.1038\/s41467-026-68351-1","volume":"17","author":"CY Lee","year":"2026","unstructured":"Lee, C. Y. et al. Central amygdala single-nucleus atlas reveals chromatin and gene transcription dynamics in human alcohol use disorder. Nat. Commun. 17, 1634 (2026).","journal-title":"Nat. Commun."},{"key":"50920_CR43","doi-asserted-by":"publisher","first-page":"1236923","DOI":"10.3389\/fimmu.2023.1236923","volume":"14","author":"W Ma","year":"2023","unstructured":"Ma, W., Huang, G., Wang, Z., Wang, L. & Gao, Q. IRF7: Role and regulation in immunity and autoimmunity. Front. Immunol. 14, 1236923 (2023).","journal-title":"Front. Immunol."},{"key":"50920_CR44","doi-asserted-by":"publisher","first-page":"eadf6484","DOI":"10.1126\/science.adf6484","volume":"382","author":"BR Lee","year":"2023","unstructured":"Lee, B. R. et al. Signature morphoelectric properties of diverse GABAergic interneurons in the human neocortex. Science 382, eadf6484 (2023).","journal-title":"Science"},{"key":"50920_CR45","doi-asserted-by":"publisher","first-page":"R909","DOI":"10.1016\/j.cub.2023.07.059","volume":"33","author":"J Zou","year":"2023","unstructured":"Zou, J. & Hires, S. A. Inhibitory neurons: VIP neurons expect rewards. Curr. Biol. 33, R909\u2013R911 (2023).","journal-title":"Curr. Biol."},{"key":"50920_CR46","doi-asserted-by":"publisher","first-page":"e12889","DOI":"10.1111\/adb.12889","volume":"26","author":"AS Warden","year":"2021","unstructured":"Warden, A. S. et al. Microglia depletion and alcohol: Transcriptome and behavioral profiles. Addict. Biol. 26, e12889 (2021).","journal-title":"Addict. Biol."},{"key":"50920_CR47","doi-asserted-by":"publisher","first-page":"101574","DOI":"10.1016\/j.arr.2022.101574","volume":"75","author":"K Ahn","year":"2022","unstructured":"Ahn, K., Lee, S-J. & Mook-Jung, I. White matter-associated microglia: New players in brain aging and neurodegenerative diseases. Ageing Res. Rev. 75, 101574 (2022).","journal-title":"Ageing Res. Rev."},{"key":"50920_CR48","doi-asserted-by":"publisher","first-page":"38","DOI":"10.1016\/j.conb.2015.09.003","volume":"36","author":"T Goldmann","year":"2016","unstructured":"Goldmann, T., Blank, T. & Prinz, M. Fine-tuning of type I IFN-signaling in microglia\u2013implications for homeostasis, CNS autoimmunity and interferonopathies. Curr. Opin. Neurobiol. 36, 38\u201342 (2016).","journal-title":"Curr. Opin. Neurobiol."},{"key":"50920_CR49","doi-asserted-by":"publisher","first-page":"500","DOI":"10.1038\/s41386-020-0721-0","volume":"46","author":"EK Erickson","year":"2021","unstructured":"Erickson, E. K. et al. Cortical astrocytes regulate ethanol consumption and intoxication in mice. Neuropsychopharmacology 46, 500\u2013508 (2021).","journal-title":"Neuropsychopharmacology"},{"key":"50920_CR50","doi-asserted-by":"publisher","first-page":"e1008142","DOI":"10.1371\/journal.ppat.1008142","volume":"15","author":"Z Luo","year":"2019","unstructured":"Luo, Z. et al. EV71 infection induces neurodegeneration via activating TLR7 signaling and IL-6 production. PLoS Pathog. 15, e1008142 (2019).","journal-title":"PLoS Pathog"},{"key":"50920_CR51","doi-asserted-by":"publisher","first-page":"576969","DOI":"10.1016\/j.jneuroim.2019.576969","volume":"333","author":"MD Rizzo","year":"2019","unstructured":"Rizzo, M. D. et al. Imiquimod and interferon-alpha augment monocyte-mediated astrocyte secretion of MCP-1, IL-6 and IP-10 in a human co-culture system. J. Neuroimmunol. 333, 576969 (2019).","journal-title":"J. Neuroimmunol."},{"key":"50920_CR52","doi-asserted-by":"publisher","first-page":"191","DOI":"10.1124\/jpet.119.260661","volume":"371","author":"MD Rizzo","year":"2019","unstructured":"Rizzo, M. D. et al. \u22069-Tetrahydrocannabinol Suppresses Monocyte-Mediated Astrocyte Production of Monocyte Chemoattractant Protein 1 and Interleukin-6 in a Toll-Like Receptor 7-Stimulated Human Coculture. J. Pharmacol. Exp. Ther. 371, 191\u2013201 (2019).","journal-title":"J. Pharmacol. Exp. Ther."},{"key":"50920_CR53","doi-asserted-by":"publisher","first-page":"740","DOI":"10.1016\/j.bbi.2020.10.007","volume":"91","author":"L Li","year":"2021","unstructured":"Li, L., Acioglu, C., Heary, R. F. & Elkabes, S. Role of astroglial toll-like receptors (TLRs) in central nervous system infections, injury and neurodegenerative diseases. Brain Behav. Immun. 91, 740\u2013755 (2021).","journal-title":"Brain Behav. Immun."},{"key":"50920_CR54","doi-asserted-by":"publisher","first-page":"627","DOI":"10.1534\/g3.111.000976","volume":"1","author":"G Joslyn","year":"2011","unstructured":"Joslyn, G. et al. Glypican gene GPC5 participates in the behavioral response to ethanol: Evidence from humans, mice, and fruit flies. G3 (Bethesda). 1, 627\u2013635 (2011).","journal-title":"G3 (Bethesda)"},{"key":"50920_CR55","doi-asserted-by":"publisher","first-page":"111","DOI":"10.1016\/j.neuint.2019.02.009","volume":"125","author":"MA Kashem","year":"2019","unstructured":"Kashem, M. A., Sultana, N., Pow, D. V. & Balcar, V. J. GLAST (GLutamate and ASpartate Transporter) in human prefrontal cortex; interactome in healthy brains and the expression of GLAST in brains of chronic alcoholics. Neurochem Int. 125, 111\u2013116 (2019).","journal-title":"Neurochem Int."},{"key":"50920_CR56","doi-asserted-by":"publisher","first-page":"105877","DOI":"10.1016\/j.phrs.2021.105877","volume":"174","author":"FE Mowry","year":"2021","unstructured":"Mowry, F. E., Peaden, S. C., Stern, J. E. & Biancardi, V. C. TLR4 and AT1R mediate blood-brain barrier disruption, neuroinflammation, and autonomic dysfunction in spontaneously hypertensive rats. Pharmacol. Res. 174, 105877 (2021).","journal-title":"Pharmacol. Res."},{"key":"50920_CR57","doi-asserted-by":"publisher","first-page":"25","DOI":"10.1186\/s12974-023-02712-1","volume":"20","author":"I Tsioti","year":"2023","unstructured":"Tsioti, I. et al. Endothelial Toll-like receptor 4 is required for microglia activation in the murine retina after systemic lipopolysaccharide exposure. J. Neuroinflammation. 20, 25 (2023).","journal-title":"J. Neuroinflammation"},{"key":"50920_CR58","first-page":"100081","volume":"8","author":"JA Sheng","year":"2024","unstructured":"Sheng, J. A., Christenson, J. R., Schwerdtfeger, L. A. & Tobet, S. A. Maternal immune activation by toll-like receptor 7 agonist during mid-gestation increases susceptibility to blood-brain barrier leakage after puberty. Brain Behav. Immun. Integr. 8, 100081 (2024).","journal-title":"Brain Behav. Immun. Integr."},{"key":"50920_CR59","doi-asserted-by":"publisher","first-page":"421","DOI":"10.1038\/s41593-024-01576-9","volume":"27","author":"C Greene","year":"2024","unstructured":"Greene, C. et al. Blood-brain barrier disruption and sustained systemic inflammation in individuals with long COVID-associated cognitive impairment. Nat. Neurosci. 27, 421\u2013432 (2024).","journal-title":"Nat. Neurosci."},{"key":"50920_CR60","doi-asserted-by":"publisher","first-page":"204173142412265","DOI":"10.1177\/20417314241226551","volume":"15","author":"L Qi","year":"2024","unstructured":"Qi, L. et al. Recent advances in tissue repair of the blood-brain barrier after stroke. J. Tissue Eng. 15, 20417314241226550 (2024).","journal-title":"J. Tissue Eng."},{"key":"50920_CR61","doi-asserted-by":"publisher","first-page":"513","DOI":"10.1016\/j.conb.2013.01.024","volume":"23","author":"J Mayfield","year":"2013","unstructured":"Mayfield, J., Ferguson, L. & Harris, R. A. Neuroimmune signaling: A key component of alcohol abuse. Curr. Opin. Neurobiol. 23, 513\u2013520 (2013).","journal-title":"Curr. Opin. Neurobiol."},{"key":"50920_CR62","doi-asserted-by":"publisher","first-page":"109400","DOI":"10.1016\/j.cbi.2021.109400","volume":"337","author":"J Wei","year":"2021","unstructured":"Wei, J. et al. Blood-brain barrier integrity is the primary target of alcohol abuse. Chem. Biol. Interact. 337, 109400 (2021).","journal-title":"Chem. Biol. Interact."},{"key":"50920_CR63","first-page":"722","volume":"57","author":"I Cetin","year":"2022","unstructured":"Cetin, I., Yazla, E., Akmese, B. & Kayadibi, H. A Preliminary Study on Investigation of Blood-Brain Barrier Damage Markers in Patients with Alcohol Use Disorder Before and After Therapy. Alcohol Alcohol. 57, 722\u2013726 (2022).","journal-title":"Alcohol Alcohol"},{"key":"50920_CR64","doi-asserted-by":"publisher","first-page":"1103","DOI":"10.1111\/adb.12376","volume":"22","author":"A Rubio-Araiz","year":"2017","unstructured":"Rubio-Araiz, A. et al. Disruption of blood-brain barrier integrity in postmortem alcoholic brain: Preclinical evidence of TLR4 involvement from a binge-like drinking model. Addict. Biol. 22, 1103\u20131116 (2017).","journal-title":"Addict. Biol."},{"key":"50920_CR65","doi-asserted-by":"publisher","first-page":"3205","DOI":"10.1242\/dev.033910","volume":"136","author":"R van Amerongen","year":"2009","unstructured":"van Amerongen, R. & Nusse, R. Towards an integrated view of Wnt signaling in development. Development 136, 3205\u20133214 (2009).","journal-title":"Development"},{"key":"50920_CR66","doi-asserted-by":"publisher","first-page":"3825","DOI":"10.1172\/JCI76431","volume":"124","author":"Y Zhou","year":"2014","unstructured":"Zhou, Y. et al. Canonical WNT signaling components in vascular development and barrier formation. J. Clin. Invest. 124, 3825\u20133846 (2014).","journal-title":"J. Clin. Invest."},{"key":"50920_CR67","doi-asserted-by":"publisher","first-page":"3","DOI":"10.1038\/s41392-021-00762-6","volume":"7","author":"J Liu","year":"2022","unstructured":"Liu, J. et al. Wnt\/\u03b2-catenin signalling: Function, biological mechanisms, and therapeutic opportunities. Signal. Transduct. Target. Ther. 7, 3 (2022).","journal-title":"Signal. Transduct. Target. Ther."},{"key":"50920_CR68","doi-asserted-by":"publisher","first-page":"103","DOI":"10.3389\/fncel.2013.00103","volume":"7","author":"SB Rosso","year":"2013","unstructured":"Rosso, S. B. & Inestrosa, N. C. WNT signaling in neuronal maturation and synaptogenesis. Front. Cell. Neurosci. 7, 103 (2013).","journal-title":"Front. Cell. Neurosci."},{"key":"50920_CR69","doi-asserted-by":"publisher","first-page":"107558","DOI":"10.1016\/j.nlm.2021.107558","volume":"187","author":"RF Narvaes","year":"2022","unstructured":"Narvaes, R. F. & Furini, C. R. G. Role of Wnt signaling in synaptic plasticity and memory. Neurobiol. Learn. Mem. 187, 107558 (2022).","journal-title":"Neurobiol. Learn. Mem."},{"key":"50920_CR70","doi-asserted-by":"publisher","first-page":"6539","DOI":"10.3390\/ijms23126539","volume":"23","author":"A Mart\u00edn-Medina","year":"2022","unstructured":"Mart\u00edn-Medina, A. et al. TLR\/WNT: A Novel Relationship in Immunomodulation of Lung Cancer. Int. J. Mol. Sci. 23, 6539 (2022).","journal-title":"Int. J. Mol. Sci."},{"key":"50920_CR71","doi-asserted-by":"publisher","first-page":"6126","DOI":"10.4049\/jimmunol.1203002","volume":"190","author":"C Oderup","year":"2013","unstructured":"Oderup, C., LaJevic, M. & Butcher, E. C. Canonical and noncanonical Wnt proteins program dendritic cell responses for tolerance. J. Immunol. 190, 6126\u20136134 (2013).","journal-title":"J. Immunol."},{"key":"50920_CR72","doi-asserted-by":"publisher","first-page":"1377501","DOI":"10.3389\/fphar.2024.1377501","volume":"15","author":"A Du","year":"2024","unstructured":"Du, A. et al. Analysis of microRNAs and the microRNA-messengerRNA regulatory network in chronic alcohol exposure. Front. Pharmacol. 15, 1377501 (2024).","journal-title":"Front. Pharmacol."},{"key":"50920_CR73","doi-asserted-by":"publisher","first-page":"103","DOI":"10.1016\/j.alcohol.2016.11.003","volume":"60","author":"R Cervera-Juanes","year":"2017","unstructured":"Cervera-Juanes, R., Wilhelm, L. J., Park, B., Grant, K. A. & Ferguson, B. Genome-wide analysis of the nucleus accumbens identifies DNA methylation signals differentiating low\/binge from heavy alcohol drinking. Alcohol 60, 103\u2013113 (2017).","journal-title":"Alcohol"},{"key":"50920_CR74","doi-asserted-by":"publisher","first-page":"e0202063","DOI":"10.1371\/journal.pone.0202063","volume":"14","author":"KM Mignogna","year":"2019","unstructured":"Mignogna, K. M., Bacanu, S. A., Riley, B. P., Wolen, A. R. & Miles, M. F. Cross-species alcohol dependence-associated gene networks: Co-analysis of mouse brain gene expression and human genome-wide association data. PLoS One. 14, e0202063 (2019).","journal-title":"PLoS One"},{"key":"50920_CR75","doi-asserted-by":"publisher","first-page":"315","DOI":"10.3233\/JAD-215508","volume":"86","author":"M Liu","year":"2022","unstructured":"Liu, M. et al. Chronic Alcohol Exposure Alters Gene Expression and Neurodegeneration Pathways in the Brain of Adult Mice. J. Alzheimers Dis. 86, 315\u2013331 (2022).","journal-title":"J. Alzheimers Dis."},{"key":"50920_CR76","doi-asserted-by":"publisher","first-page":"e25118","DOI":"10.1371\/journal.pone.0025118","volume":"6","author":"RG Govindaraj","year":"2011","unstructured":"Govindaraj, R. G., Manavalan, B., Basith, S. & Choi, S. Comparative analysis of species-specific ligand recognition in Toll-like receptor 8 signaling: A hypothesis. PLoS One. 6, e25118 (2011).","journal-title":"PLoS One"},{"issue":"Suppl 1","key":"50920_CR77","doi-asserted-by":"publisher","first-page":"S92","DOI":"10.1016\/j.bbi.2011.01.008","volume":"25","author":"YA Blednov","year":"2011","unstructured":"Blednov, Y. A. et al. Activation of inflammatory signaling by lipopolysaccharide produces a prolonged increase of voluntary alcohol intake in mice. Brain Behav. Immun. 25 (Suppl 1), S92\u2013S105 (2011).","journal-title":"Brain Behav. Immun."},{"key":"50920_CR78","doi-asserted-by":"publisher","first-page":"293","DOI":"10.1038\/s41587-023-01767-y","volume":"42","author":"Y Hao","year":"2024","unstructured":"Hao, Y. et al. Dictionary learning for integrative, multimodal and scalable single-cell analysis. Nat. Biotechnol. 42, 293\u2013304 (2024).","journal-title":"Nat. Biotechnol."},{"key":"50920_CR79","doi-asserted-by":"publisher","first-page":"296","DOI":"10.1186\/s13059-019-1874-1","volume":"20","author":"C Hafemeister","year":"2019","unstructured":"Hafemeister, C. & Satija, R. Normalization and variance stabilization of single-cell RNA-seq data using regularized negative binomial regression. Genome Biol. 20, 296 (2019).","journal-title":"Genome Biol."},{"key":"50920_CR80","doi-asserted-by":"publisher","first-page":"69","DOI":"10.1186\/s13059-021-02281-7","volume":"22","author":"H Guo","year":"2021","unstructured":"Guo, H. & Li, J. scSorter: Assigning cells to known cell types according to marker genes. Genome Biol. 22, 69 (2021).","journal-title":"Genome Biol."},{"key":"50920_CR81","doi-asserted-by":"publisher","first-page":"4720","DOI":"10.1093\/bioinformatics\/btac582","volume":"38","author":"B Phipson","year":"2022","unstructured":"Phipson, B. et al. Propeller: Testing for differences in cell type proportions in single cell data. Bioinformatics 38, 4720\u20134726 (2022).","journal-title":"Bioinformatics"},{"key":"50920_CR82","doi-asserted-by":"publisher","first-page":"5692","DOI":"10.1038\/s41467-021-25960-2","volume":"12","author":"JW Squair","year":"2021","unstructured":"Squair, J. W. et al. Confronting false discoveries in single-cell differential expression. Nat. Commun. 12, 5692 (2021).","journal-title":"Nat. Commun."},{"key":"50920_CR83","doi-asserted-by":"publisher","first-page":"9588","DOI":"10.1038\/s41598-018-27903-2","volume":"8","author":"KM Cahill","year":"2018","unstructured":"Cahill, K. M., Huo, Z., Tseng, G. C., Logan, R. W. & Seney, M. L. Improved identification of concordant and discordant gene expression signatures using an updated rank-rank hypergeometric overlap approach. Sci. Rep. 8, 9588 (2018).","journal-title":"Sci. Rep."},{"key":"50920_CR84","doi-asserted-by":"publisher","first-page":"100498","DOI":"10.1016\/j.crmeth.2023.100498","volume":"3","author":"S Morabito","year":"2023","unstructured":"Morabito, S., Reese, F., Rahimzadeh, N., Miyoshi, E. & Swarup, V. hdWGCNA identifies co-expression networks in high-dimensional transcriptomics data. Cell. Rep. Methods. 3, 100498 (2023).","journal-title":"Cell. Rep. Methods"},{"key":"50920_CR85","doi-asserted-by":"publisher","first-page":"3573","DOI":"10.1016\/j.cell.2021.04.048","volume":"184","author":"Y Hao","year":"2021","unstructured":"Hao, Y. et al. Integrated analysis of multimodal single-cell data. Cell 184, 3573\u20133587e29 (2021).","journal-title":"Cell"},{"key":"50920_CR86","doi-asserted-by":"publisher","first-page":"3223","DOI":"10.1093\/bioinformatics\/bty332","volume":"34","author":"Z Miao","year":"2018","unstructured":"Miao, Z., Deng, K., Wang, X. & Zhang, X. DEsingle for detecting three types of differential expression in single-cell RNA-seq data. Bioinformatics 34, 3223\u20133224 (2018).","journal-title":"Bioinformatics"}],"container-title":["Scientific Reports"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.nature.com\/articles\/s41598-026-50920-5","content-type":"text\/html","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/www.nature.com\/articles\/s41598-026-50920-5.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/www.nature.com\/articles\/s41598-026-50920-5.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,7,1]],"date-time":"2026-07-01T08:31:23Z","timestamp":1782894683000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.nature.com\/articles\/s41598-026-50920-5"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,4,30]]},"references-count":86,"journal-issue":{"issue":"1","published-online":{"date-parts":[[2026,12]]}},"alternative-id":["50920"],"URL":"https:\/\/doi.org\/10.1038\/s41598-026-50920-5","relation":{},"ISSN":["2045-2322"],"issn-type":[{"value":"2045-2322","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,4,30]]},"assertion":[{"value":"25 November 2025","order":1,"name":"received","label":"Received","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"24 April 2026","order":2,"name":"accepted","label":"Accepted","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"30 April 2026","order":3,"name":"first_online","label":"First Online","group":{"name":"ArticleHistory","label":"Article History"}},{"order":1,"name":"Ethics","group":{"name":"EthicsHeading","label":"Declarations"}},{"value":"The authors declare no competing interests.","order":2,"name":"Ethics","group":{"name":"EthicsHeading","label":"Competing interests"}}],"article-number":"20115"}}