Fig. 1

Figure 1. Expression of Dendra- α-SYN in transgenic zebrafish. (A) Representative brightfield and fluorescence images of Dendra-α-SYN positive offspring from stable UAS:Dendra-α-SYN-wt and A53T fish lines crossed to the PanN:Gal4VP16 driver fish at 1, 2 and 3 d.p.f. Expression localised to the CNS remains strong from 1 to 3 d.p.f. and does not result in morphological defects. Scale bar represents 1 mm. (B) Quantification by qPCR of the relative expression levels of Dendra/GAPDH in offspring from selected Dendra-α-SYN-wt (black) and A53T founder fish (white) showing comparable mRNA levels at 1, 2 and 3 d.p.f. when outcrossed to pan-neuronal or ubiquitous GAL4 driver lines (PanN:GAL4-VP16 and ubb:GAL4-VP16). Both groups showed reduced Dendra mRNA levels over time normalised by GAPDH. Data represents mean ± s.d. (N = 3 independent clutches for each age and genotype) (C) Western blot for total α-SYN and phosphorylated α-SYN at residue S129 (pS129) in whole fish lysates from 3 d.p.f. fish with pan-neuronal expression of Dendra-α-SYN-wt and A53T compared to tubulin levels as loading control. (D) Densitometry of total α-SYN and pS129 immunoblots in (C). The two transgenic lines had equivalent α-SYN levels relative to tubulin, whereas the levels of pS129 α-SYN were significantly increased in Dendra-α-SYN-A53T mutant fish, compared to α-SYN-wt (** p < 0.01 vs. α-SYN-wt).