Exposure to copper is correlated with increased neutrophils in the peripheral and central nervous system. (A–C) Ventral views of whole mount adult brains from Tg(mpx:GFP). Scale bars: A-C; A’-C’= 100 μm. (A, A’) Control with neutrophils found only in OO (arrowhead; A’). (B, B’, B”) After four-hour exposure to copper, there was an increase in the number of neutrophils in the OO (B, arrowhead, B’, B”). In addition, neutrophils were observed in the ventral OB, along the ventricle (V) and in the ventral telencephalon (B, arrows) (C, C’) One day post treatment neutrophils were still present in OO (C’), the OB (arrows) and the ventral telencephalon. (D) Neutrophils appear over time in an anterior to posterior spatial pattern: olfactory nerve (ON), olfactory bulb (OB), posterior Telencephalon (pTel), and anterior diencephalon; (the numbers for OO are not plotted because number is out of range (average ~1,500, see E). (E) Copper exposure was correlated with increased neutrophils in OE and ventral CNS. ****: Two-way ANOVA, Tukey’s multiple comparison test, p < 0.05). (****P, 0.0001). (A–C, A’–C’) Preparations were selected for imaging based on whether they were intact and the signal to noise of the labeling. (D) 1 brain was analyzed per timepoint. (E) 3 brains were examined per treatment.
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