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Yet, the mechanism underlying SD-evoked neuroinflammation and trigeminovascular activation remains incompletely understood.<\/jats:p>\n                  <jats:p>We characterized the identity of inflammasome activated following SD-evoked PANX1 opening. Pharmacological inhibitors targeting PANX1 or NLRP3 as well as genetic ablation of Nlrp3 and Il1b were applied to investigate the molecular mechanism of the downstream neuroinflammatory cascades. In addition, we examined whether SD-triggered microglial activation facilitates neuronal NLRP3-mediated inflammatory cascades. Pharmacological inhibition of toll-like receptors TLR2\/4, the potential receptors of the damage-associated molecular pattern HMGB1, was further employed to interrogate the neuron-microglia interplay in SD-induced neuroinflammation.<\/jats:p>\n                  <jats:p>We found that NLRP3 but not NLRP1 or NLRP2 inflammasome was activated following PANX1 opening after single or multiple SDs evoked by either KCl topical application or non-invasively with optogenetics. The SD-evoked NLRP3 inflammasome activation was observed exclusively in neurons but not microglia or astrocytes. Proximity ligation assay demonstrated that the assembly of the NLRP3 inflammasome occurred as early as 15 min after SD. Genetic ablation of Nlrp3 or Il1b or pharmacological inhibition of PANX1 or NLRP3 ameliorated SD-induced neuronal inflammation, middle meningeal artery dilatation, calcitonin gene-related peptide expression in trigeminal ganglion and c-Fos expression in trigeminal nucleus caudalis. Moreover, multiple SDs induced microglial activation subsequent to neuronal NLRP3 inflammasome activation, which in turn orchestrated with neurons to mediate cortical neuroinflammation, as demonstrated by decreased neuronal inflammation after pharmacological inhibition of microglia activation or blockade of the TLR2\/4 receptors.<\/jats:p>\n                  <jats:p>To conclude, single or multiple SDs evoked activation of neuronal NLRP3 inflammasomes and its downstream inflammatory cascades to mediate cortical neuroinflammation and trigeminovascular activation. In the context of multiple SDs, the cortical inflammatory processes could be facilitated by SD-evoked microglia activation. These findings may implicate the potential role of innate immunity in migraine pathogenesis.<\/jats:p>","DOI":"10.1093\/brain\/awad045","type":"journal-article","created":{"date-parts":[[2023,2,15]],"date-time":"2023-02-15T00:50:28Z","timestamp":1676422228000},"page":"2989-3002","source":"Crossref","is-referenced-by-count":68,"title":["Neuronal NLRP3 inflammasome mediates spreading depolarization-evoked trigeminovascular activation"],"prefix":"10.1093","volume":"146","author":[{"given":"Pin-Yu","family":"Chen","sequence":"first","affiliation":[{"name":"Institute of Clinical Medicine, College of Medicine, National Yang Ming Chiao Tung University , Taipei 112 , Taiwan"},{"name":"Faculty of Medicine, College of Medicine, National Yang Ming Chiao Tung University , Taipei 112 , Taiwan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jiin-Cherng","family":"Yen","sequence":"additional","affiliation":[{"name":"Institute of Pharmacology, College of Medicine, National Yang Ming Chiao Tung University , Taipei 112 , Taiwan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Tzu-Ting","family":"Liu","sequence":"additional","affiliation":[{"name":"Institute of Pharmacology, College of Medicine, National Yang Ming Chiao Tung University , Taipei 112 , Taiwan"},{"name":"Brain Research Center, National Yang Ming Chiao Tung University , Taipei 112 , Taiwan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Szu-Ting","family":"Chen","sequence":"additional","affiliation":[{"name":"Institute of Clinical Medicine, College of Medicine, National Yang Ming Chiao Tung University , Taipei 112 , Taiwan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4488-9310","authenticated-orcid":false,"given":"Shuu-Jiun","family":"Wang","sequence":"additional","affiliation":[{"name":"Faculty of Medicine, College of Medicine, National Yang Ming Chiao Tung University , Taipei 112 , Taiwan"},{"name":"Brain Research Center, National Yang Ming Chiao Tung University , Taipei 112 , Taiwan"},{"name":"Department of Neurology, Neurological Institute, Taipei Veterans General Hospital , Taipei 112 , Taiwan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3492-9902","authenticated-orcid":false,"given":"Shih-Pin","family":"Chen","sequence":"additional","affiliation":[{"name":"Institute of Clinical Medicine, College of Medicine, National Yang Ming Chiao Tung University , Taipei 112 , Taiwan"},{"name":"Faculty of Medicine, College of Medicine, National Yang Ming Chiao Tung University , Taipei 112 , Taiwan"},{"name":"Brain Research Center, National Yang Ming Chiao Tung University , Taipei 112 , Taiwan"},{"name":"Department of Neurology, Neurological Institute, Taipei Veterans General Hospital , Taipei 112 , Taiwan"},{"name":"Division of Translational Research, Department of Medical Research, Taipei Veterans General Hospital , Taipei 112 , 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