Figure 6

Heat shock induced Wnt8a overexpression affects hypothalamic DC5 and DC6 DA neurons in a stage dependent manner. (a–l) Expression of DA neuron marker th as detected by WISH in WT (a,c,e,g,i,k) and in heterozygous transgenic Tg(hsp70l:wnt8a-GFP) siblings (b,d,f,h,j,l) subjected to heat-shock treatments as indicated on the left and subsequently fixed at 80 hpf. Dorsal views, images are Z-projections of image stacks. The th expressing DC groups 2, 4, 5 and 6 are indicated in (a), (b), (c) and (d). The arrowheads in (d), (f), (h) and (j) point to an increase in th expression in DC5 and DC6 DA neuron groups. Scale bar in (b) is 100 µm for all images. Numbers N/N in bottom right corner of each image indicate number of representative phenotypes as shown in image versus total embryos analyzed for this condition. (m) A separate experiment was performed for quantification of TH-immunoreactive cells in WT embryos compared to heterozygous transgenic Tg(hsp70l:wnt8a-GFP) siblings heat shocked as indicated and fixed at 82 hpf. Bar charts show cell count numbers and the mean of TH-immunoreactive cells for each indicated DA neuron groups. Error bars depict standard deviations of the mean. Asterisks indicate significant differences compared to non-transgenic sibling control (**p = 0.001 for 15 hpf and p = 0.0009 for 18 hpf; Mann–Whitney U test; n = 14 for 15 hpf, n = 9 for 18 hpf and n = 8 for 25 hpf. Software: Prism 6.0f. from GraphPad Software Inc.).