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Phenotype of neuronal loss in LRRK2 ΔWD40 morphants. (A) Western blot analysis at 2 dpf and 3 dpf confirming the knockdown effect of zLRRK2 wild-type protein expression as well as the decreased TH protein expression in WD40 deletion morphant. Adult brain protein was used as positive control, and the ATG morphant at 3 dpf was used as negative control. The quantified results are shown in Figure S2. (B) WISH analysis at 3 dpf showing that WD40 domain deletion caused decreased TH and DAT expressions in WD40 deletion morphant, but without significant brain developmental retardation. (C) Uninjected (left) and WD40 deleted zebrafish expressing Kaede under the deltaD promoter. While neuron numbers is not much changed at 18 somite stage, there are fewer neurons in the midbrain of these 6 dpf fish, as indicated by the arrowheads. ***P<0.001, n.s.: not significant (unpaired Student′s t-test). (D) Cell apoptosis assay of uninjected (left) and WD40 delete zebrafish embryos.
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