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In some pathological conditions, microglia produce increased levels of cytotoxic and inflammatory mediators, such as tumor necrosis factor alpha (TNF-\u03b1), which can reactivate microglia in a positive feedback mechanism. However, specific molecular mediators that can be effectively targeted to control TNF-\u03b1-mediated microglia overactivation, are yet to be uncovered. In this context, we aim to identify novel TNF-\u03b1-mediated micro(mi)RNAs and to dissect their roles in microglia activation, as well as to explore their impact on the cellular communication with neurons. A miRNA microarray, followed by RT-qPCR validation, was performed on TNF-\u03b1-stimulated primary rat microglia. Gain- and loss-of-function in vitro assays and proteomic analysis were used to dissect the role of miR-342 in microglia activation. Co-cultures of microglia with hippocampal neurons, using a microfluidic system, were performed to understand the impact on neurotoxicity. Stimulation of primary rat microglia with TNF-\u03b1 led to an upregulation of <jats:italic>Nos2<\/jats:italic>, <jats:italic>Tnf<\/jats:italic>, and <jats:italic>Il1b<\/jats:italic> mRNAs. In addition, ph-NF-kB p65 levels were also increased. miRNA microarray analysis followed by RT-qPCR validation revealed that TNF-\u03b1 stimulation induced the upregulation of miR-342. Interestingly, miR-342 overexpression in N9 microglia was sufficient to activate the NF-kB pathway by inhibiting BAG-1, leading to increased secretion of TNF-\u03b1 and IL-1\u03b2. Conversely, miR-342 inhibition led to a strong decrease in the levels of these cytokines after TNF-\u03b1 activation. In fact, both TNF-\u03b1-stimulated and miR-342-overexpressing microglia drastically affected neuron viability. Remarkably, increased levels of nitrites were detected in the supernatants of these co-cultures. Globally, our findings show that miR-342 is a crucial mediator of TNF-\u03b1-mediated microglia activation and a potential target to tackle microglia-driven neuroinflammation.<\/jats:p>","DOI":"10.1038\/s41419-020-2626-6","type":"journal-article","created":{"date-parts":[[2020,6,2]],"date-time":"2020-06-02T10:05:50Z","timestamp":1591092350000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":194,"title":["TNF-alpha-induced microglia activation requires miR-342: impact on NF-kB signaling and neurotoxicity"],"prefix":"10.1038","volume":"11","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-1867-0486","authenticated-orcid":false,"given":"Jo\u00e3o Paulo","family":"Br\u00e1s","sequence":"first","affiliation":[]},{"given":"Joana","family":"Bravo","sequence":"additional","affiliation":[]},{"given":"Jaime","family":"Freitas","sequence":"additional","affiliation":[]},{"given":"M\u00e1rio Adolfo","family":"Barbosa","sequence":"additional","affiliation":[]},{"given":"Susana Gomes","family":"Santos","sequence":"additional","affiliation":[]},{"ORCID":"https:\/\/orcid.org\/0000-0003-2548-6281","authenticated-orcid":false,"given":"Teresa","family":"Summavielle","sequence":"additional","affiliation":[]},{"ORCID":"https:\/\/orcid.org\/0000-0003-2072-8587","authenticated-orcid":false,"given":"Maria In\u00eas","family":"Almeida","sequence":"additional","affiliation":[]}],"member":"297","published-online":{"date-parts":[[2020,6,2]]},"reference":[{"key":"2626_CR1","doi-asserted-by":"publisher","first-page":"22","DOI":"10.1038\/nri.2015.5","volume":"16","author":"AH Miller","year":"2016","unstructured":"Miller, A. 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