{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,22]],"date-time":"2026-10-22T06:39:16Z","timestamp":1792651156992,"version":"4.5.0"},"reference-count":36,"publisher":"MDPI AG","issue":"11","license":[{"start":{"date-parts":[[2024,5,22]],"date-time":"2024-05-22T00:00:00Z","timestamp":1716336000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"National Natural Science Foundation of China","award":["32070161"],"award-info":[{"award-number":["32070161"]}]},{"name":"National Natural Science Foundation of China","award":["2022YFD1800100"],"award-info":[{"award-number":["2022YFD1800100"]}]},{"name":"National Natural Science Foundation of China","award":["2020GXNSFAA297212"],"award-info":[{"award-number":["2020GXNSFAA297212"]}]},{"name":"National Natural Science Foundation of China","award":["31902311"],"award-info":[{"award-number":["31902311"]}]},{"name":"National Key Research and Development Program of China","award":["32070161"],"award-info":[{"award-number":["32070161"]}]},{"name":"National Key Research and Development Program of China","award":["2022YFD1800100"],"award-info":[{"award-number":["2022YFD1800100"]}]},{"name":"National Key Research and Development Program of China","award":["2020GXNSFAA297212"],"award-info":[{"award-number":["2020GXNSFAA297212"]}]},{"name":"National Key Research and Development Program of China","award":["31902311"],"award-info":[{"award-number":["31902311"]}]},{"name":"Natural Science Foundation of Guangxi Province, China","award":["32070161"],"award-info":[{"award-number":["32070161"]}]},{"name":"Natural Science Foundation of Guangxi Province, China","award":["2022YFD1800100"],"award-info":[{"award-number":["2022YFD1800100"]}]},{"name":"Natural Science Foundation of Guangxi Province, China","award":["2020GXNSFAA297212"],"award-info":[{"award-number":["2020GXNSFAA297212"]}]},{"name":"Natural Science Foundation of Guangxi Province, China","award":["31902311"],"award-info":[{"award-number":["31902311"]}]},{"name":"National Natural Science Foundation of China","award":["32070161"],"award-info":[{"award-number":["32070161"]}]},{"name":"National Natural Science Foundation of China","award":["2022YFD1800100"],"award-info":[{"award-number":["2022YFD1800100"]}]},{"name":"National Natural Science Foundation of China","award":["2020GXNSFAA297212"],"award-info":[{"award-number":["2020GXNSFAA297212"]}]},{"name":"National Natural Science Foundation of China","award":["31902311"],"award-info":[{"award-number":["31902311"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>Rabies virus (RABV) is a neurotropic virus that causes fatal neurological disease, raising serious public health issues and attracting extensive attention in society. To elucidate the molecular mechanism of RABV-induced neuronal damage, we used hematoxylin\u2013eosin staining, transmission electron microscopy, transcriptomics analysis, and immune response factor testing to investigate RABV-infected neurons. We successfully isolated the neurons from murine brains. The specificity of the isolated neurons was identified by a monoclonal antibody, and the viability of the neurons was 83.53\u201395.0%. We confirmed that RABV infection induced serious damage to the neurons according to histochemistry and transmission electron microscope (TEM) scanning. In addition, the transcriptomics analysis suggested that multiple genes related to the pyroptosis pathway were significantly upregulated, including gasdermin D (Gsdmd), Nlrp3, caspase-1, and IL-1\u03b2, as well as the chemokine genes Ccl2, Ccl3, Ccl4, Ccl5, Ccl7, Ccl12, and Cxcl10. We next verified this finding in the brains of mice infected with the rRC-HL, GX074, and challenge virus standard strain-24 (CVS-24) strains of RABV. Importantly, we found that the expression level of the Gsdmd protein was significantly upregulated in the neurons infected with different RABV strains and ranged from 691.1 to 5764.96 pg\/mL, while the basal level of mock-infected neurons was less than 100 pg\/mL. Taken together, our findings suggest that Gsdmd-induced pyroptosis is involved in the neuron damage caused by RABV infection.<\/jats:p>","DOI":"10.3390\/ijms25115616","type":"journal-article","created":{"date-parts":[[2024,5,22]],"date-time":"2024-05-22T07:56:11Z","timestamp":1716364571000},"page":"5616","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":15,"title":["Rabies Virus Infection Causes Pyroptosis of Neuronal Cells"],"prefix":"10.3390","volume":"25","author":[{"given":"Dongling","family":"Yu","sequence":"first","affiliation":[{"name":"College of Animal Science and Veterinary Medicine, Guangxi University, Nanning 530004, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Rong","family":"Jin","sequence":"additional","affiliation":[{"name":"College of Animal Science and Veterinary Medicine, Guangxi University, Nanning 530004, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jundan","family":"Liu","sequence":"additional","affiliation":[{"name":"College of Animal Science and Veterinary Medicine, Guangxi University, Nanning 530004, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Chuanliang","family":"Zhang","sequence":"additional","affiliation":[{"name":"College of Animal Science and Veterinary Medicine, Guangxi University, Nanning 530004, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Chenxing","family":"Duan","sequence":"additional","affiliation":[{"name":"College of Animal Science and Veterinary Medicine, Guangxi University, Nanning 530004, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xi","family":"Luo","sequence":"additional","affiliation":[{"name":"College of Animal Science and Veterinary Medicine, Guangxi University, Nanning 530004, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Wenhao","family":"Yang","sequence":"additional","affiliation":[{"name":"College of Animal Science and Veterinary Medicine, Guangxi University, Nanning 530004, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Cheng","family":"Liu","sequence":"additional","affiliation":[{"name":"College of Animal Science and Veterinary Medicine, Guangxi University, Nanning 530004, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4623-9864","authenticated-orcid":false,"given":"Jingjing","family":"Liang","sequence":"additional","affiliation":[{"name":"College of Animal Science and Veterinary Medicine, Guangxi University, Nanning 530004, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xiaoning","family":"Li","sequence":"additional","affiliation":[{"name":"College of Animal Science and Veterinary Medicine, Guangxi University, Nanning 530004, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Tingrong","family":"Luo","sequence":"additional","affiliation":[{"name":"College of Animal Science and Veterinary Medicine, Guangxi University, Nanning 530004, China"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2024,5,22]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"1389","DOI":"10.1016\/S0140-6736(13)62707-5","article-title":"Current Status of Rabies and Prospects for Elimination","volume":"384","author":"Fooks","year":"2014","journal-title":"Lancet"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"61","DOI":"10.1016\/j.antiviral.2013.01.003","article-title":"Current and Future Approaches to the Therapy of Human Rabies","volume":"99","author":"Jackson","year":"2013","journal-title":"Antivir. Res."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"485","DOI":"10.1016\/0042-6822(90)90433-R","article-title":"Molecular Cloning and Complete Nucleotide Sequence of the Attenuated Rabies Virus SAD B19","volume":"175","author":"Conzelmann","year":"1990","journal-title":"Virology"},{"key":"ref_4","doi-asserted-by":"crossref","unstructured":"Koraka, P., Martina, B.E.E., van den Ham, H.-J., Zaaraoui-Boutahar, F., van IJcken, W., Roose, J., van Amerongen, G., Andeweg, A., and Osterhaus, A.D.M.E. (2018). Analysis of Mouse Brain Transcriptome After Experimental Duvenhage Virus Infection Shows Activation of Innate Immune Response and Pyroptotic Cell Death Pathway. Front. Microbiol., 9.","DOI":"10.3389\/fmicb.2018.00397"},{"key":"ref_5","doi-asserted-by":"crossref","unstructured":"Hampson, K., Coudeville, L., Lembo, T., Sambo, M., Kieffer, A., Attlan, M., Barrat, J., Blanton, J.D., Briggs, D.J., and Cleaveland, S. (2015). Estimating the Global Burden of Endemic Canine Rabies. PLoS Negl. Trop. Dis., 9.","DOI":"10.1371\/journal.pntd.0003786"},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"629","DOI":"10.1080\/14760584.2019.1627205","article-title":"Current Status of Human Rabies Prevention: Remaining Barriers to Global Biologics Accessibility and Disease Elimination","volume":"18","author":"Rupprecht","year":"2019","journal-title":"Expert Rev. Vaccines"},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"146","DOI":"10.1186\/s12974-014-0146-y","article-title":"Met-CCL5 Represents an Immunotherapy Strategy to Ameliorate Rabies Virus Infection","volume":"11","author":"Huang","year":"2014","journal-title":"J. Neuroinflamm."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"169","DOI":"10.1016\/j.virusres.2005.08.004","article-title":"Multiple Amino Acids in the Glycoprotein of Rabies Virus Are Responsible for Pathogenicity in Adult Mice","volume":"115","author":"Ito","year":"2006","journal-title":"Virus Res."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"8926","DOI":"10.1128\/JVI.00787-10","article-title":"Molecular Basis of Neurovirulence of Flury Rabies Virus Vaccine Strains: Importance of the Polymerase and the Glycoprotein R333Q Mutation","volume":"84","author":"Tao","year":"2010","journal-title":"J. Virol."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"89","DOI":"10.1111\/j.1348-0421.2009.00192.x","article-title":"Amino Acid Substitutions at Positions 242, 255 and 268 in Rabies Virus Glycoprotein Affect Spread of Viral Infection","volume":"54","author":"Ito","year":"2010","journal-title":"Microbiol. Immunol."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"e00810-22","DOI":"10.1128\/jvi.00810-22","article-title":"The Amino Acid at Position 95 in the Matrix Protein of Rabies Virus Is Involved in Antiviral Stress Granule Formation in Infected Cells","volume":"96","author":"Kojima","year":"2022","journal-title":"J. Virol."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"418","DOI":"10.1002\/ame2.12273","article-title":"Regulation of Innate Immune Responses by Rabies Virus","volume":"5","author":"Zhang","year":"2022","journal-title":"Anim. Models Exp. Med."},{"key":"ref_13","doi-asserted-by":"crossref","unstructured":"Feige, L., Kozaki, T., Dias De Melo, G., Guillemot, V., Larrous, F., Ginhoux, F., and Bourhy, H. (2022). Susceptibilities of CNS Cells towards Rabies Virus Infection Is Linked to Cellular Innate Immune Responses. Viruses, 15.","DOI":"10.3390\/v15010088"},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"1704","DOI":"10.1080\/15548627.2016.1196315","article-title":"Wild-Type Rabies Virus Induces Autophagy in Human and Mouse Neuroblastoma Cell Lines","volume":"12","author":"Peng","year":"2016","journal-title":"Autophagy"},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"637","DOI":"10.1016\/j.it.2015.08.002","article-title":"Immune Surveillance of the CNS Following Infection and Injury","volume":"36","author":"Russo","year":"2015","journal-title":"Trends Immunol."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"11808","DOI":"10.1128\/JVI.01346-09","article-title":"The Roles of Chemokines in Rabies Virus Infection: Overexpression May Not Always Be Beneficial","volume":"83","author":"Zhao","year":"2009","journal-title":"J. Virol."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"18","DOI":"10.1016\/j.virusres.2009.03.014","article-title":"Role of Chemokines in the Enhancement of BBB Permeability and Inflammatory Infiltration after Rabies Virus Infection","volume":"144","author":"Kuang","year":"2009","journal-title":"Virus Res."},{"key":"ref_18","doi-asserted-by":"crossref","unstructured":"Scott, T.P., and Nel, L.H. (2016). Subversion of the Immune Response by Rabies Virus. Viruses, 8.","DOI":"10.3390\/v8080231"},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"486","DOI":"10.1038\/s41418-017-0012-4","article-title":"Molecular Mechanisms of Cell Death: Recommendations of the Nomenclature Committee on Cell Death 2018","volume":"25","author":"Galluzzi","year":"2018","journal-title":"Cell Death Differ."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"245","DOI":"10.1016\/j.tibs.2016.10.004","article-title":"Pyroptosis: Gasdermin-Mediated Programmed Necrotic Cell Death","volume":"42","author":"Shi","year":"2017","journal-title":"Trends Biochem. Sci."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"212","DOI":"10.1080\/01652176.2017.1343516","article-title":"Rabies\u2014Epidemiology, Pathogenesis, Public Health Concerns and Advances in Diagnosis and Control: A Comprehensive Review","volume":"37","author":"Singh","year":"2017","journal-title":"Vet. Q."},{"key":"ref_22","first-page":"749","article-title":"Comparative Analysis of the Pathogenic Mechanisms of Street Rabies Virus Strains with Different Virulence Levels","volume":"27","author":"Yin","year":"2014","journal-title":"Biomed. Environ. Sci."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"154","DOI":"10.1111\/j.1865-1682.2011.01255.x","article-title":"Pathological, Immunological and Molecular Diagnosis of Rabies in Clinically Suspected Animals of Different Species Using Four Detection Techniques in Jordan: Pathological, Immunological and Molecular Diagnosis of Rabies","volume":"59","author":"Faizee","year":"2012","journal-title":"Transbound. Emerg. Dis."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"630","DOI":"10.1177\/0300985810370013","article-title":"Immunohistochemical Study of Rabies Virus within the Central Nervous System of Domestic and Wildlife Species","volume":"47","author":"Stein","year":"2010","journal-title":"Vet. Pathol."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"518","DOI":"10.1080\/135502801753248105","article-title":"Silver-Haired Bat Rabies Virus Variant Does Not Induce Apoptosis in the Brain of Experimentally Infected Mice","volume":"7","author":"Yan","year":"2001","journal-title":"J. Neurovirol."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"2163","DOI":"10.1007\/s11250-020-02239-0","article-title":"Molecular Characterization and Diagnostic Investigations of Rabies Encephalitis in Camels (Camelus Dromedaries) in Oman: A Retrospective Study","volume":"52","author":"Ahmed","year":"2020","journal-title":"Trop. Anim. Health Prod."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"157","DOI":"10.1186\/s12985-020-01429-4","article-title":"Inflammatory Response and MAPK and NF-\u03baB Pathway Activation Induced by Natural Street Rabies Virus Infection in the Brain Tissues of Dogs and Humans","volume":"17","author":"Liu","year":"2020","journal-title":"Virol. J."},{"key":"ref_28","doi-asserted-by":"crossref","unstructured":"Liu, J., Li, W., Yu, D., Jin, R., Hou, H., Ling, X., Kiflu, A.B., Wei, X., Yang, X., and Li, X. (2023). Transcriptomic Analysis of mRNA Expression Profiles in the Microglia of Mouse Brains Infected with Rabies Viruses of Varying Virulence. Viruses, 15.","DOI":"10.3390\/v15061223"},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"942","DOI":"10.1128\/CMR.17.4.942-964.2004","article-title":"Role of Microglia in Central Nervous System Infections","volume":"17","author":"Rock","year":"2004","journal-title":"Clin. Microbiol. Rev."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"4681","DOI":"10.1016\/j.vaccine.2018.04.002","article-title":"Inhibition of Caspase-1 Prolongs Survival of Mice Infected with Rabies Virus","volume":"37","author":"Koraka","year":"2019","journal-title":"Vaccine"},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"4701","DOI":"10.1016\/j.vaccine.2018.04.003","article-title":"The Effect of Combined Drugs Therapy on the Course of Clinical Rabies Infection in a Murine Model","volume":"37","author":"Smreczak","year":"2019","journal-title":"Vaccine"},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"4736","DOI":"10.1016\/j.vaccine.2018.05.065","article-title":"Combination Drug Treatment Prolongs Survival of Experimentally Infected Mice with Silver-Haired Bat Rabies Virus","volume":"37","author":"Martina","year":"2019","journal-title":"Vaccine"},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"51","DOI":"10.1111\/j.1348-0421.2001.tb01274.x","article-title":"A Comparison of Complete Genome Sequences of the Attenuated RC-HL Strain of Rabies Virus Used for Production of Animal Vaccine in Japan, and the Parental Nishigahara Strain","volume":"45","author":"Ito","year":"2001","journal-title":"Microbiol. Immunol."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"561","DOI":"10.17221\/63\/2018-VETMED","article-title":"The N and P Genes Facilitate Pathogenicity of the Rabies Virus G Gene","volume":"63","author":"Wei","year":"2018","journal-title":"Vet. Med."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"463","DOI":"10.7883\/yoken.64.463","article-title":"Gene expression analysis of host innate immune responses in the central nervous system following lethal CVS-11 infection in mice","volume":"64","author":"Sugiura","year":"2011","journal-title":"Jpn. J. Infect. Dis."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"30529","DOI":"10.1038\/srep30529","article-title":"Viperin Inhibits Rabies Virus Replication via Reduced Cholesterol and Sphingomyelin and Is Regulated Upstream by TLR4","volume":"6","author":"Tang","year":"2016","journal-title":"Sci. Rep."}],"container-title":["International Journal of Molecular Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1422-0067\/25\/11\/5616\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,10]],"date-time":"2025-10-10T14:46:07Z","timestamp":1760107567000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1422-0067\/25\/11\/5616"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2024,5,22]]},"references-count":36,"journal-issue":{"issue":"11","published-online":{"date-parts":[[2024,6]]}},"alternative-id":["ijms25115616"],"URL":"https:\/\/doi.org\/10.3390\/ijms25115616","relation":{},"ISSN":["1422-0067"],"issn-type":[{"value":"1422-0067","type":"electronic"}],"subject":[],"published":{"date-parts":[[2024,5,22]]}}}