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Neurosci."],"published-print":{"date-parts":[[2020,10,7]]},"abstract":"<jats:p>Microglia, a type of CNS immune cell, have been shown to contribute to ethanol-activated neuronal death of the stress regulatory proopiomelanocortin (POMC) neuron-producing \u03b2-endorphin peptides in the hypothalamus in a postnatal rat model of fetal alcohol spectrum disorders. We determined whether the microglial extracellular vesicle exosome is involved in the ethanol-induced neuronal death of the \u03b2-endorphin neuron. Extracellular vesicles were prepared from hypothalamic tissues collected from postnatal rats (both males and females) fed daily with 2.5 mg\/kg ethanol or control milk formula for 5 d or from hypothalamic microglia cells obtained from postnatal rats, grown in cultures for several days, and then challenged with ethanol or vehicle for 24 h. Nanoparticle tracking analysis and transmission electron microscopy indicated that these vesicles had the size range and shape of exosomes. Ethanol treatments increased the number and the \u03b2-endorphin neuronal killing activity of microglial exosomes both<jats:italic>in vivo<\/jats:italic>and<jats:italic>in vitro<\/jats:italic>. Proteomics analyses of exosomes of cultured microglial cells identified a large number of proteins, including various complements, which were elevated following ethanol treatment. Proteomics data involving complements were reconfirmed using quantitative protein assays. Ethanol treatments also increased deposition of the complement protein C1q in \u03b2-endorphin neuronal cells in both<jats:italic>in vitro<\/jats:italic>and<jats:italic>in vivo<\/jats:italic>systems. Recombinant C1q protein increased while C1q blockers reduced ethanol-induced C3a\/b, C4, and membrane attack complex\/C5b9 formations; ROS production; and ultimately cellular death of \u03b2-endorphin neurons. These data suggest that the complement system involving C1q-C3-C4-membrane attack complex and ROS regulates exosome-mediated, ethanol-induced \u03b2-endorphin neuronal death.<\/jats:p><jats:p><jats:bold>SIGNIFICANCE STATEMENT<\/jats:bold>Neurotoxic action of alcohol during the developmental period is recognized for its involvement in fetal alcohol spectrum disorders, but the lack of clear understanding of the mechanism of alcohol action has delayed the progress in therapeutic intervention of this disease. Proopiomelanocortin neurons known to regulate stress, energy homeostasis, and immune functions are reported to be killed by developmental alcohol exposure because of activation of microglial immune cells in the brain. While microglia are known to use extracellular vesicles to communicate with neurons for maintaining homeostasis, we show here that ethanol exposure during the developmental period hijacks this system to spread apoptotic factors, including complement protein C1q, to induce the membrane attack complex and reactive super-oxygen species for proopiomelanocortin neuronal killing.<\/jats:p>","DOI":"10.1523\/jneurosci.0284-20.2020","type":"journal-article","created":{"date-parts":[[2020,9,4]],"date-time":"2020-09-04T21:50:16Z","timestamp":1599256216000},"page":"7965-7979","source":"Crossref","is-referenced-by-count":58,"title":["Alcohol Increases Exosome Release from Microglia to Promote Complement C1q-Induced Cellular Death of Proopiomelanocortin Neurons in the Hypothalamus in a Rat Model of Fetal Alcohol Spectrum 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