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Fig. 2. `Comma-shaped' neurogenic areas in the juvenile crayfish brain. (A) Diagram
of the juvenile crayfish brain identifying the neuropils and cell clusters
[modified from Sandeman et al. (Sandeman
et al., 1992)] as well as the neurogenic areas in the
deutocerebrum (this study). Primary sensory inputs from olfactory receptor
neurons (ORN) in olfactory organs enter the olfactory lobe (OL). The cell
bodies of the olfactory local and projection interneurons comprise cell
clusters 9 and 10, respectively. Groups of BrdU+ cells (green) appear in cell
clusters 9 and 10 and lines of BrdU+ cells, surrounded with a strand-like
structure (violet), meet at a patch of cells (blue) near the posterior
accessory lobe (AL). 9, cell cluster 9; 10, cell cluster 10; AMPN, anterior
median protocerebral neuropil; AN, antennal neuropil; DCN, deutocerebral
commissure neuropil; LAN, lateral antennular neuropil; PMPN, posterior median
protocerebral neuropil. (B) Juvenile crayfish brain showing robust cell
proliferation in cell clusters 9 and 10. The image is taken with the camera
attached to the epifluorescence microscope. Each dotted area (orange for
cluster 9 head and white for cluster 10 head) indicates regions where a ball
of BrdU+ cells is present within the cluster boundary. Each broken area
indicates a region around the LAN (orange for cluster 9 tail) or around the
posterior AL (white for cluster 10 tail) where `tails' of BrdU+ cells are
located. Scale bar, 50 µm. (C–F) Collapsed stacks of confocal images
taken at 1.0 µm intervals in (C) and 0.5 µm intervals in (D–F).
The entire stack for each image was approximately 60 µm. (C) A strand
labeled with anti-tubulin (violet, arrows), surrounds lines of BrdU+ cells
(green) from clusters 9 and 10, and a patch of cells (arrowhead). Scale bar,
50 µm. (D) High-resolution image of the patch of cells (green, anti-BrdU;
blue, Hoechst, nuclear staining; violet, anti-tubulin) indicated by an
arrowhead in (C). The patch of cells and a few BrdU+ cells are surrounded by a
tubulin+ strand. Scale bar, 50 µm. (E) Lines of BrdU+ cells (green) in the
tail of cluster 9 are surrounded by a strand faintly labeled with anti-H3P
(red, arrows). BrdU+ cells outside the strand (arrowheads) were not included
in our counts of labeled cells. Asterisks indicate H3P+ (mitotic) cells. Scale
bar, 50 µm. (F) Lines of BrdU+ cells (green) in the tail of cluster 10 are
surrounded by a strand faintly labeled with anti-H3P (red, arrows). A BrdU+
cell on the border of AL (arrowhead) was not included in our counts of labeled
cell. Asterisk indicates H3P+ (mitotic) cells. Scale bar, 50 µm.
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