Microglia response to brain injury following optogenetic silencing of neurons in PFOS-exposed larvae. (A) Confocal micrograph of a 3-dpf control-treated larval brain with fluorescently labeled microglia in green and halorhodopsin+ neurons in magenta. Image taken 4 hpi without light stimulation. (B) Isolated microglia from (A), pseudocolored in black and white. Area of microglia response 4 hpi shaded with magenta. (C) Confocal micrograph of a 3-dpf control-treated larval brain with fluorescently labeled microglia in green and halorhodopsin+ neurons in magenta. Image taken 4 hpi following stimulation with 589-nm light. (D) Isolated microglia from (C), pseudocolored in black and white. (E) Confocal micrograph of a 3-dpf 28μM PFOS-treated larval brain with fluorescently labeled microglia in green and halorhodopsin+ neurons in magenta. Image taken 4 hpi without light stimulation. (F) Isolated microglia from (E), pseudocolored in black and white. (G) Confocal micrograph of a 3-dpf 28μM-treated larval brain with fluorescently labeled microglia in green and halorhodopsin+ neurons in magenta. Image taken 4 hpi following stimulation with 589-nm light. (H) Isolated microglia from (G), pseudocolored in black and white. (I) Quantification of the area of microglia response around the injury site at 4 hpi, as shown by the magenta overlays. n=14–20 fish per group. Error bars represent standard deviation. Box plot limits represent 25th to 75th percentile, with the midline representing the median. See Excel Table S1 for additional statistical details. Note: dpf, days postfertilization; hpi, hours post-injury; ns, not significant; PFOS, perfluorooctane sulfonate.
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