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Fig. 3. Fate mapping of the diencephalic progenitor zones. High-resolution
fate mapping by two-photon-based uncaging procedure.
(A,A') Zebrafish embryos (anterior to the left) expressing
GFP under the control of the neurog1 promoter (neurog1::gfp)
were injected with caged dextran-fluorescein tracer dye at the 1-cell stage.
At the 1- to 3-somite stage, the dye was uncaged at the indicated domains
(denoted I, II and III) of the diencephalic anlage. (A) Dorsal view;
(A') lateral view. (B) Schematic summarizing the results of
multiple uncaging experiments showing the final destination of the
fluorescein-labeled cells in 24-hpf neurog1::gfp embryos. The clones
corresponding to each of the uncaged domains are color coded (I, n=6;
II, n=6; III, n=10). (C-D'') Control (WT;
C-C'') and dkk1 mRNA-injected (D-D'') embryos that
underwent uncaging were fixed at 24 hpf, followed by immunofluorescence
staining of the uncaged fluorescein and of TH+ DA neurons.
High-magnification images of a diencephalic area (dashed boxes in C,D)
containing neurog1+ TH+ fluorescein+
triple-positive cells (arrowheads) are shown in
C',C'',D',D''. Dien, diencephalon; Tel, telencephalon;
TG, trigeminal ganglion; VCC, ventrocaudal cluster. Scale bars: 25 µm in
A-D; 50 µm in C',C'',D',D''.
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