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Fig. 4
Zebrafish GDNF overexpression prevented human 4R-tau-GFP and zebrafish 3R-tau-GFP induced Neuronal death. a GFP-labeled neuronal cells and axons were observed at 24 and 48 hpf in embryos co-injected with pGFAP-zGDNF-HA and pHuC-z3R-tau-GFP (b) or pHuC-h4R-tau-GFP (d). For comparison, embryos co-injected with pGFAP-zBDNF-HA (panels a and c) were used as the control. The boxed regions are enlarged (a′-d′′) to show the GFP-labeled neuronal cells in 24 to 48 hpf embryos from the lateral view. Scale bars: 100 µm. The protection effect of GDNF against neuronal death induced by human tau-GFP or zebrafish tau-GFP was presented in panel e to show higher percentage, 55 % and 53 % of zebrafish embryos expressing GDNF with more neuronal cells, compared to 20 % and 19 % without GDNF. b Double immunostaining of h4R-tau-GFP (GFP antibody, panel a) and GDNF-HA (HA antibody, panel a′) in spinal cord neurons of the aforementioned zebrafish embryos. The phosphorylation state of h4R-tau-GFP was detected using antibody pT212 (panel b) and antibody AT8 (panel b′). Scale bar: 50 µm