Fig. 3

The effect of H₂O₂ on GFP and TH1 expression in Tg(pink1:EGFP) larval fish.

A-C. GFP-ir at 7 dpf in the zebrafish larvae. GFP-ir increases in the H₂O₂-treated fish (B) as compared to the control fish (A). The effect is rescued in the LGR treated fish (C).

D. Mean fluorescence intensity values of the WT, H₂O₂-treated group and the H₂O₂+LGR treated group. A significant change in the H₂O₂-treated group was observed (p < 0.01).

E-G. A three-dimensional image to perform volume rendering was constructed using the Imaris MeasurementPro module in Imaris software for the above-mentioned fish at a similar threshold intensity. The changes indicated as the changed total volume of the representative images for control (E), H₂O₂ (F) and LGR-treated fish (G). Scale bar represents 80 μm.

H-J. TH-ir is unchanged among the control (H), H₂O₂-treated (I) and H₂O₂ together with LGR (J) groups of larval fish. No change in the pretectal (7) and preoptic, thalamic or pre-tectal nucleus (5,6,11) cell populations was seen. Other visible groups in the olfactory bulb and ventral telencephalic nuclei (1,2), periventricular hypothalamus and posterior tuberculum (13), and the locus coeruleus (14) also displayed no change in TH-ir.

Scale bar represents 100 μm. A-C and H-J. H2O2 – Hydrogen Peroxide, GFP – Green fluorescent protein, ir – immunoreactivity, LGR – L-glutathione reduced, PINK1 – PTEN-induced putative kinase 1, TH – tyrosine hydroxylase.