Light/dark behavioral assay and neuronal activity in microglia mutants exposed to PFOS. Wild-type (WT) and microglia-deficient irf8st96/st96 larvae dosed with either control or 7μM PFOS were assessed for potential changes in swim behavior and neuronal activity. (A) Light/dark behavioral assay of 5-dpf WT or irf8−/− mutant control or PFOS-exposed larvae. (B) Assessment of the average distance moved of 5-dpf larvae at minute 15, which is the start of the second light-off cycle, at minute 30, which is the end of the second light-off cycle, and the percentage change in distance moved from minute 15 to minute 30. (C) Quantification of neuronal CaMPARI activity in the forebrain of 5-dpf control-treated WT, PFOS-treated WT, control-treated mutant, and PFOS-treated mutant larvae. (D) Quantification of neuronal CaMPARI activity in the optic tectum. (E) Quantification of neuronal CaMPARI activity in the cerebellum. (F) Quantification of neuronal CaMPARI activity in the hindbrain. (G) Quantification of neuronal CaMPARI activity in the whole brain. n=14–22 fish per group for behavior; n=13–16 fish per group for neuroimaging. **p<0.01; ***p<0.001; ****p<0.0001. 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: CaMPARI, calcium-modulated photoactivatable ratiometric integrator; Ce, cerebellum; dpf, days postfertilization; FB, forebrain; HB, hindbrain; OT, optic tectum; PFOS, perfluorooctane sulfonate; WB, whole brain.
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