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Fig. 8. Non-specifying adenohypophyseal cells of fgf3 mutants undergo
apoptosis. (A-D,I,J) Nomarski images, (G,H) confocal images; frontal views
onto the anterior border of the head, superimposed with fluorescent images of
Acridine Orange stainings in green (C,D,J), or rhodamine cell labelings in red
(G-J). All images are at same magnification; scale bar shown in (I) = 25
µm. In A-D,I, borders of the adenohypophysis are indicated with white
arrowheads. All embryos were genotyped after evaluation and photography. (A)
Nomarski images at 25 hpf; wild-type sibling; (B) fgf3 mutant,
displaying a normal-sized adenohypophysis. For better contrast, images are not
superimposed with Acridine Orange stainings, which showed no positive cells
for wild-type (A), but few positive cells for mutant (B). (C,D) Acridine
Orange stainings at 28 hpf; (C) wild-type sibling; (D) fgf3 mutant.
(E,F) Summary of single cell-tracing experiments: cartoons showing the
position of single cells from the anterior neural ridge region labeled at the
tailbud stage, relative to mesodermal polster outlined in gray, which served
as a landmark for the injections. A square corresponds to 50 µm x 50
µm. The fate of each cell is indicated by shape and color. Circles mark
cells whose daughter cells were alive 28-30 hpf, crosses indicate cells that
died during the course of the analysis, resulting in labeled cell debris only.
Red circles mark adenohypophyseal clones, blue circles olfactory epithelium
clones, yellow circles facial ectoderm clones, and salmon circles clones
ending up in the head mesenchyme. Mixed clones with descendants in two
different tissues are indicated with two-colored circles. Numbers and arrows
mark cells with descendants shown in G-J. (E) Cells from wild-type siblings;
(F) cells from fgf3 mutants. In F, red crosses represent clones with
debris within and outside the shrunken adenohypophysis. Gray crosses are
clones with debris outside the adenohypophysis only, which had not been
investigated at 24 hpf. Therefore, their initial tissue belongings cannot be
stated; however, they most likely derive from adenohypophyseal cells that had
died early. (G) Descendants of wild-type cell 54 at 29 hpf. (H) Cell debris
deriving from mutant cell 49. (I,J) Time course analyses of mutant cell 84.
(I) 25 hpf; (H) same embryo at 30 hpf, counterstained with Acridine
Orange.
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