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Fig. 5. Larval melanocyte regeneration following MoTP treatment is achieved by
cell division. (A-H) Cell division events in melanocyte lineages
during larval melanocyte regeneration were tracked by BrdU incorporation
experiments. Larvae were continuously incubated in BrdU (5 mM) during and
after MoTP treatment, then fixed, paraffin embedded and 5 µm sagittal
sections processed for BrdU immunohistochemistry after melanocyte regeneration
was mostly completed at 10 dpf (5 days post-MoTP treatment). BrdU
incorporation states of pigmented melanocytes were assessed by first
identifying melanocyte nuclei (thinning in melanin, white arrowheads in A and
E) accompanied by bisbenzimide staining (white arrowheads in B and F). These
were then examined for red fluorescence indicative of BrdU incorporation
(white arrowheads in C and G). D is the overlay image of A, B and C,
indicating a melanocyte that did not incorporate BrdU during the BrdU
labeling, whereas in H, the overlay image of D, E and F shows a different
melanocyte that did incorporate BrdU during the BrdU labeling. (I)
Quantitative analyses of BrdU incorporation in larvae exposed to MoTP from 4
to 5 dpf. Black and white bars indicate BrdU incorporation in untreated and
MoTP-treated larvae, respectively. Horizontal green line indicates periods of
BrdU labeling. Asterisk in I indicates the developmental stages at which
larvae were sacrificed for BrdU incorporation analysis. Scale bar in A: 10
µm for A-H.
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