{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,11]],"date-time":"2026-09-11T12:08:30Z","timestamp":1789128510069,"version":"build-2803163510"},"reference-count":50,"publisher":"MDPI AG","issue":"10","license":[{"start":{"date-parts":[[2020,5,20]],"date-time":"2020-05-20T00:00:00Z","timestamp":1589932800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100004837","name":"Ministerio de Ciencia e Innovaci\u00f3n","doi-asserted-by":"publisher","award":["BFU2014-53641-P and BFU2017-85723-P"],"award-info":[{"award-number":["BFU2014-53641-P and BFU2017-85723-P"]}],"id":[{"id":"10.13039\/501100004837","id-type":"DOI","asserted-by":"publisher"}]},{"name":"Junta de Extremadura.","award":["GR18185, AA-17-0099-1 and IDA1-18-0041-3"],"award-info":[{"award-number":["GR18185, AA-17-0099-1 and IDA1-18-0041-3"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>3-Nitropropionic acid (NPA) administration to rodents produces degeneration of the striatum, accompanied by neurological disturbances that mimic Huntington\u2019s disease (HD) motor neurological dysfunctions. It has been shown that inflammation mediates NPA-induced brain degeneration, and activated microglia secreting cytokines interleukin-1\u03b1 (IL-1\u03b1) and tumor necrosis factor \u03b1 (TNF\u03b1) can induce a specific type of reactive neurotoxic astrocytes, named A1, which have been detected in post-mortem brain samples of Huntington\u2019s, Alzheimer\u2019s, and Parkinson\u2019s diseases. In this work we used an experimental model based on the intraperitoneal (i.p.) administration of NPA to adult Wistar rats at doses that can elicit extensive brain degeneration, and brain samples were taken before and after extensive brain damage monitored using 2,3,5-triphenyltetrazolium chloride (TTC) staining. Western blots and immunohistochemistry of brain slices show that i.p. NPA injections elicit significant increase in the expression levels of C3\u03b1 subunit, a marker of generation of neurotoxic A1 astrocytes, and of cytokines IL-1\u03b1, TNF\u03b1, and C1q within the striatum, hippocampus, and cerebellum before the appearance of the HD-related neurological dysfunctions and neuronal death induced by NPA. Noteworthy, NPA administration primarily induces the generation of A1 astrocytes in the more recent phylogenetic area of the rat cerebellum. We conclude that the activation of complement C3 protein in the brain from Wistar rats is an early event in NPA-induced brain neurodegeneration.<\/jats:p>","DOI":"10.3390\/ijms21103609","type":"journal-article","created":{"date-parts":[[2020,5,20]],"date-time":"2020-05-20T10:37:38Z","timestamp":1589971058000},"page":"3609","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":32,"title":["Early Reactive A1 Astrocytes Induction by the Neurotoxin 3-Nitropropionic Acid in Rat Brain"],"prefix":"10.3390","volume":"21","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-3296-2071","authenticated-orcid":false,"given":"Carmen","family":"Lopez-Sanchez","sequence":"first","affiliation":[{"name":"Instituto de Biomarcadores de Patologias Moleculares, Universidad de Extremadura, 06006 Badajoz, Spain"},{"name":"Departamento de Anatomia y Embriologia Humana, Facultad de Medicina, Universidad de Extremadura, 06006 Badajoz, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7760-1985","authenticated-orcid":false,"given":"Virginio","family":"Garcia-Martinez","sequence":"additional","affiliation":[{"name":"Instituto de Biomarcadores de Patologias Moleculares, Universidad de Extremadura, 06006 Badajoz, Spain"},{"name":"Departamento de Anatomia y Embriologia Humana, Facultad de Medicina, Universidad de Extremadura, 06006 Badajoz, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Joana","family":"Poejo","sequence":"additional","affiliation":[{"name":"Instituto de Biomarcadores de Patologias Moleculares, Universidad de Extremadura, 06006 Badajoz, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1489-7296","authenticated-orcid":false,"given":"Virginio","family":"Garcia-Lopez","sequence":"additional","affiliation":[{"name":"Instituto de Biomarcadores de Patologias Moleculares, Universidad de Extremadura, 06006 Badajoz, Spain"},{"name":"Departamento de Anatomia y Embriologia Humana, Facultad de Medicina, Universidad de Extremadura, 06006 Badajoz, Spain"},{"name":"Departamento de Enfermeria, Facultad de Medicina, Universidad de Extremadura, 06006 Badajoz, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jairo","family":"Salazar","sequence":"additional","affiliation":[{"name":"Instituto de Biomarcadores de Patologias Moleculares, Universidad de Extremadura, 06006 Badajoz, Spain"},{"name":"Departamento de Quimica, Universidad Nacional Autonoma de Nicaragua-Leon, Leon 21000, Nicaragua"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3673-7007","authenticated-orcid":false,"given":"Carlos","family":"Gutierrez-Merino","sequence":"additional","affiliation":[{"name":"Instituto de Biomarcadores de Patologias Moleculares, Universidad de Extremadura, 06006 Badajoz, Spain"},{"name":"Departamento de Bioquimica y Biolog\u00eda Molecular y Genetica, Facultad de Ciencias, Universidad de Extremadura, 06006 Badajoz, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2020,5,20]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"3767","DOI":"10.1073\/pnas.74.9.3767","article-title":"3-Nitropropionate, the toxic substance of Indigofera, is a suicide inactivator of succinate dehydrogenase","volume":"74","author":"Alston","year":"1977","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_2","first-page":"161","article-title":"Studies on the epidemiology and etiology of moldy sugarcane poisoning in China","volume":"5","author":"Liu","year":"1992","journal-title":"Biomed. Environ. Sci."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"363","DOI":"10.3109\/15563659509028924","article-title":"Moldy Sugarcane Poisoning\u2014A Case Report with a Brief Review","volume":"33","author":"Ming","year":"1995","journal-title":"J. Toxicol. Clin. Toxicol."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"214","DOI":"10.1016\/j.yrtph.2003.09.002","article-title":"Production of toxic metabolites in Aspergillus niger, Aspergillus oryzae, and Trichoderma reesei: Justification of mycotoxin testing in food grade enzyme preparations derived from the three fungi","volume":"39","author":"Blumenthal","year":"2004","journal-title":"Regul. Toxicol. Pharmacol."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"4181","DOI":"10.1523\/JNEUROSCI.13-10-04181.1993","article-title":"Neurochemical and histologic characterization of striatal excitotoxic lesions produced by the mitochondrial toxin 3-nitropropionic acid","volume":"13","author":"Beal","year":"1993","journal-title":"J. Neurosci."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"542","DOI":"10.1097\/00019052-199412000-00012","article-title":"Neurochemistry and toxin models in Huntington\u02bcs disease","volume":"7","author":"Beal","year":"1994","journal-title":"Curr. Opin. Neurol."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"356","DOI":"10.1111\/j.1471-4159.1993.tb05859.x","article-title":"Age-Dependent Vulnerability of the Striatum to the Mitochondrial Toxin 3-Nitropropionic Acid","volume":"60","author":"Brouillet","year":"1993","journal-title":"J. Neurochem."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"427","DOI":"10.1016\/S0301-0082(99)00005-2","article-title":"Replicating Huntington\u2019s disease phenotype in experimental animals","volume":"59","author":"Brouillet","year":"1999","journal-title":"Prog. Neurobiol."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"1769","DOI":"10.1097\/00001756-200106130-00050","article-title":"Topological analysis of striatal lesions induced by 3-nitropropionic acid in the Lewis rat","volume":"12","author":"Blum","year":"2001","journal-title":"NeuroReport"},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"473","DOI":"10.1111\/j.1471-4159.2009.06331.x","article-title":"Kaempferol protects against rat striatal degeneration induced by 3-nitropropionic acid","volume":"111","author":"Lagoa","year":"2009","journal-title":"J. Neurochem."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"559","DOI":"10.1097\/00005072-198511000-00003","article-title":"Neuropathological classification of Huntington\u2019s disease","volume":"44","author":"Vonsattel","year":"1985","journal-title":"J. Neuropathol. Exp. Neurol."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"1702","DOI":"10.1212\/01.WNL.0000098878.47789.BD","article-title":"Profile of cognitive progression in early Huntington\u2019s disease","volume":"61","author":"Ho","year":"2003","journal-title":"Neurology"},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"1491","DOI":"10.1002\/mds.21971","article-title":"The current clinical management of Huntington\u2019s disease","volume":"23","author":"Phillips","year":"2008","journal-title":"Mov. Disord."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"147","DOI":"10.1111\/j.1750-3639.1999.tb00216.x","article-title":"Oxidative stress in Huntington\u2019s disease","volume":"9","author":"Browne","year":"1999","journal-title":"Brain Pathol."},{"key":"ref_15","doi-asserted-by":"crossref","unstructured":"Menze, E., Esmat, A., Tadros, M.G., Abdel-Naim, A.B., and Khalifa, A.E. (2015). Genistein Improves 3-NPA-Induced Memory Impairment in Ovariectomized Rats: Impact of Its Antioxidant, Anti-Inflammatory and Acetylcholinesterase Modulatory Properties. PLoS ONE, 10.","DOI":"10.1371\/journal.pone.0117223"},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"1261","DOI":"10.1007\/s11064-008-9904-5","article-title":"Metabonomic Characterization of the 3-Nitropropionic Acid Rat Model of Huntington\u2019s Disease","volume":"34","author":"Tsang","year":"2009","journal-title":"Neurochem. Res."},{"key":"ref_17","doi-asserted-by":"crossref","unstructured":"Peraza, A.V., Guzm\u00e1n, D.C., Brizuela, N.O., Herrera, M.O., Olguin, H.J., Silva, M.L., Tapia, B.J., and Mej\u00eda, G.B. (2018). Riboflavin and pyridoxine restore dopamine levels and reduce oxidative stress in brain of rats. BMC Neurosci., 19.","DOI":"10.1186\/s12868-018-0474-4"},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"396","DOI":"10.1002\/ana.25413","article-title":"Cerebellar degeneration correlates with motor symptoms in Huntington disease","volume":"85","author":"Mehrabi","year":"2019","journal-title":"Ann. Neurol."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"89","DOI":"10.1016\/S0197-0186(02)00229-2","article-title":"Influence of cytosolic and mitochondrial Ca2+, ATP, mitochondrial membrane potential, and calpain activity on the mechanism of neuron death induced by 3-nitropropionic acid","volume":"43","author":"Nasr","year":"2003","journal-title":"Neurochem. Int."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"1220","DOI":"10.1046\/j.1471-4159.2003.02250.x","article-title":"Mitochondrial calcium, oxidative stress and apoptosis in a neurodegenerative disease model induced by 3-nitropropionic acid","volume":"88","author":"Rosenstock","year":"2004","journal-title":"J. Neurochem."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"1521","DOI":"10.1111\/j.1471-4159.2005.03515.x","article-title":"3-Nitropropionic acid: A mitochondrial toxin to uncover physiopathological mechanisms underlying striatal degeneration in Huntington\u2019s disease","volume":"95","author":"Brouillet","year":"2005","journal-title":"J. Neurochem."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"2803","DOI":"10.1111\/j.1460-9568.2008.06310.x","article-title":"Mechanisms of neurodegeneration in Huntington\u2019s disease","volume":"27","author":"Gil","year":"2008","journal-title":"Eur. J. Neurosci."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"455","DOI":"10.1046\/j.1471-4159.1995.64010455.x","article-title":"NMDA receptor involvement in toxicity to dopamine neurons in vitro caused by the succinate dehydrogenase inhibitor 3-nitropropionic acid","volume":"64","author":"Zeevalk","year":"1995","journal-title":"J. Neurochem."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"503","DOI":"10.1016\/S0306-4522(97)00389-8","article-title":"3-Nitropropionic acid exacerbates N-methyl-d-aspartate toxicity in striatal culture by multiple mechanisms","volume":"84","author":"Greene","year":"1998","journal-title":"Neuroscience"},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"5110","DOI":"10.1523\/JNEUROSCI.21-14-05110.2001","article-title":"Inhibition of Mitochondrial Complex II Induces a Long-Term Potentiation of NMDA-Mediated Synaptic Excitation in the Striatum Requiring Endogenous Dopamine","volume":"21","author":"Calabresi","year":"2001","journal-title":"J. Neurosci."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"5020","DOI":"10.1523\/JNEUROSCI.23-12-05020.2003","article-title":"Calpain Is a Major Cell Death Effector in Selective Striatal Degeneration Induced In Vivo by 3-Nitropropionate: Implications for Huntington\u2019s Disease","volume":"23","author":"Bizat","year":"2003","journal-title":"J. Neurosci."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"77","DOI":"10.1016\/j.neulet.2007.05.038","article-title":"3-Nitropropionic acid activates calpain\/cdk5 pathway in rat striatum","volume":"421","author":"Camins","year":"2007","journal-title":"Neurosci. Lett."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"333","DOI":"10.1016\/0892-0362(94)00076-P","article-title":"3-Nitropropionic acid produces indirect excitotoxic damage to rat striatum","volume":"17","author":"Fu","year":"1995","journal-title":"Neurotoxicol. Teratol."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"694","DOI":"10.1046\/j.1471-4159.2000.0750694.x","article-title":"Astrocyte-Derived Nitric Oxide Causes Both Reversible and Irreversible Damage to the Neuronal Mitochondrial Respiratory Chain","volume":"75","author":"Stewart","year":"2002","journal-title":"J. Neurochem."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"121","DOI":"10.1016\/S0969-9961(03)00002-0","article-title":"Microglial activation and cell death induced by the mitochondrial toxin 3-nitropropionic acid: In vitro and in vivo studies","volume":"12","author":"Ryu","year":"2003","journal-title":"Neurobiol. Dis."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"10","DOI":"10.1111\/cns.12189","article-title":"Quercetin Improves Behavioral Deficiencies, Restores Astrocytes and Microglia, and Reduces Serotonin Metabolism in 3-Nitropropionic Acid-Induced Rat Model of Huntington\u2019s Disease","volume":"20","author":"Chakraborty","year":"2013","journal-title":"CNS Neurosci. Ther."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"5942","DOI":"10.1038\/s41598-018-24385-0","article-title":"Morphological characterization of NG2 glia and their association with neuroglial cells in the 3-nitropropionic acid\u2013lesioned striatum of rat","volume":"8","author":"Jin","year":"2018","journal-title":"Sci. Rep."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"301","DOI":"10.4329\/wjr.v6.i6.301","article-title":"Neuroimaging in Huntington\u2019s disease","volume":"6","author":"Niccolini","year":"2014","journal-title":"World J. Radiol."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"S2","DOI":"10.5213\/inj.1632604.302","article-title":"Pathophysiological Role of Neuroinflammation in Neurodegenerative Diseases and Psychiatric Disorders","volume":"20","author":"Hong","year":"2016","journal-title":"Int. Neurourol. J."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"21362","DOI":"10.1074\/jbc.M600504200","article-title":"Tumor Necrosis Factor-\u03b1 Induces Neurotoxicity via Glutamate Release from Hemichannels of Activated Microglia in an Autocrine Manner","volume":"281","author":"Takeuchi","year":"2006","journal-title":"J. Biol. Chem."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"364","DOI":"10.1016\/j.it.2015.04.007","article-title":"The choreography of neuroinflammation in Huntington\u2019s disease","volume":"36","author":"Crotti","year":"2015","journal-title":"Trends Immunol."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"343","DOI":"10.1016\/S0168-0102(97)01170-X","article-title":"Acute 3-nitropropionic acid intoxication induces striatal astrocytic cell death and dysfunction of the blood-brain barrier: Involvement of dopamine toxicity","volume":"27","author":"Nishino","year":"1997","journal-title":"Neurosci. Res."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"481","DOI":"10.1038\/nature21029","article-title":"Neurotoxic reactive astrocytes are induced by activated microglia","volume":"541","author":"Liddelow","year":"2017","journal-title":"Nature"},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"11929","DOI":"10.1523\/JNEUROSCI.1860-14.2014","article-title":"An RNA-sequencing transcriptome and splicing database of glia, neurons, and vascular cells of the cerebral cortex","volume":"34","author":"Zhang","year":"2014","journal-title":"J. Neurosci."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"E1738","DOI":"10.1073\/pnas.1525528113","article-title":"New tools for studying microglia in the mouse and human CNS","volume":"113","author":"Bennett","year":"2016","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"362","DOI":"10.1006\/exnr.1999.7170","article-title":"Increased Complement Biosynthesis By Microglia and Complement Activation on Neurons in Huntington\u2019s Disease","volume":"159","author":"Singhrao","year":"1999","journal-title":"Exp. Neurol."},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"644","DOI":"10.1002\/jnr.21875","article-title":"C1q, the recognition subcomponent of the classical pathway of complement, drives microglial activation","volume":"87","author":"Cheung","year":"2009","journal-title":"J. Neurosci. Res."},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"265","DOI":"10.1006\/exnr.1995.1029","article-title":"DNA End Labeling (TUNEL) in Huntington\u2019s Disease and Other Neuropathological Conditions","volume":"133","author":"Thomas","year":"1995","journal-title":"Exp. Neurol."},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"35","DOI":"10.1016\/j.neulet.2006.09.037","article-title":"Occurrence of complement protein C3 in dying pyramidal neurons in rat hippocampus after systemic administration of kainic acid","volume":"409","author":"Morita","year":"2006","journal-title":"Neurosci. Lett."},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"276","DOI":"10.1186\/s12974-016-0742-0","article-title":"Complement component 3 (C3) expression in the hippocampus after excitotoxic injury: Role of C\/EBP\u03b2","volume":"13","author":"Gine","year":"2016","journal-title":"J. Neuroinflamm."},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"250","DOI":"10.1016\/0006-8993(92)91004-X","article-title":"Microglial-produced nitric oxide and reactive nitrogen oxides mediate neuronal cell death","volume":"587","author":"Boje","year":"1992","journal-title":"Brain Res."},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"1662","DOI":"10.3389\/fncel.2015.00018","article-title":"Interleukin-1 and acute brain injury","volume":"9","author":"Murray","year":"2015","journal-title":"Front. Cell. Neurosci."},{"key":"ref_48","doi-asserted-by":"crossref","first-page":"244","DOI":"10.1186\/s12974-015-0458-6","article-title":"Ligand engagement of Toll-like receptors regulates their expression in cortical microglia and astrocytes","volume":"12","author":"Marinelli","year":"2015","journal-title":"J. Neuroinflamm."},{"key":"ref_49","doi-asserted-by":"crossref","first-page":"521","DOI":"10.1016\/S0306-4522(00)00020-8","article-title":"Major strain differences in response to chronic systemic administration of the mitochondrial toxin 3-nitropropionic acid in rats: Implications for neuroprotection studies","volume":"97","author":"Ouary","year":"2000","journal-title":"Neuroscience"},{"key":"ref_50","doi-asserted-by":"crossref","first-page":"82","DOI":"10.1016\/j.jneumeth.2005.05.008","article-title":"Transfemoral selective \u201cintraluminal wiring\u201d technique for transient middle cerebral artery occlusion in rats","volume":"149","author":"Sun","year":"2005","journal-title":"J. Neurosci. Methods"}],"container-title":["International Journal of Molecular Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1422-0067\/21\/10\/3609\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T09:30:47Z","timestamp":1760175047000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1422-0067\/21\/10\/3609"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2020,5,20]]},"references-count":50,"journal-issue":{"issue":"10","published-online":{"date-parts":[[2020,5]]}},"alternative-id":["ijms21103609"],"URL":"https:\/\/doi.org\/10.3390\/ijms21103609","relation":{},"ISSN":["1422-0067"],"issn-type":[{"value":"1422-0067","type":"electronic"}],"subject":[],"published":{"date-parts":[[2020,5,20]]}}}