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Fig. 2. Coordinated expression of pac1, fezl and Otp in the NPO and PT
areas. (A-D,F) Sagittal paraffin sections (6 µm) through
the NPO and PT (lateral view, anterior to the left). At the indicated stages
of development, embryos were subjected to whole-mount in situ hybridization
with probes directed against pac1 (A,B), fezl (C,D) or
isotocin (it; F). Thereafter, specimens were sectioned and
immunostained with an anti-Otp antibody. Schemes of fezl expression
domain (dark purple) at the plane of the section are shown in C and D. Labeled
RNA probes were detected in the cytoplasm, whereas Otp antibody exclusively
labeled the nuclei. (E) Projected confocal Z-stack images of
embryos (lateral view, anterior to the left) that were subjected to double
immunofluorescence staining with monoclonal and polyclonal antibodies directed
against tyrosine hydroxylase (TH) and Otp, respectively. The two prominent
clusters of dopaminergic neurons, group 2 (Gr. 2) and groups 3-6 (Gr. 3-6),
are indicated. hpf, hours post fertilization; NPO, neurosecretory preoptic
area; PT, posterior tuberculum; vDien, ventral diencephalon. Scale bars: 20
µm in A-D; 40 µm in E,F.
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