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Fig. 1. Neural plate olig2:EGFP+ precursors generate PMNs
and KA' interneurons at the same time. (A) Dorsal view
of the posterior neural plate at 11.5 hpf (five-somite stage) and (B-F)
lateral view of 6- to 12-somite regions of spinal cord at 25 hpf in
Tg(olig2:egfp) zebrafish embryos. (A) Heterogeneous primary neuronal
populations within the EGFP+ domain. Asterisks and arrowheads mark
EGFP+ Hu+ Isl+ PMNs and EGFP+
Hu+ Isl- interneurons, respectively. Isl protein is
nuclear, whereas Hu is cytoplasmic. (B-F) Embryos were incubated with BrdU at
successive timepoints (as shown to the left of each panel) and labeled with
anti-GABA (red) and anti-BrdU (blue) antibodies. Arrows and arrowheads mark
BrdU+ and BrdU- KA' interneurons, respectively.
(B) Four KA' interneurons were formed from EGFP+ precursors
that underwent S phase at 10 hfp (arrows). Arrowhead marks BrdU-
KA' interneuron, indicating that the postmitotic cell was formed before
or after 10 hpf. (C) Two BrdU+ KA' interneurons (arrows) and
two BrdU- KA' interneurons (arrowheads) were detected in the
embryos that were incubated with BrdU at 11 hpf. (D,E,F) Embryos incubated
with BrdU at 12, 14 and 18 hpf. No BrdU+ KA' interneurons
were evident. (G) Average of all KA' cells (squares) versus
BrdU+ KA' cells (triangles) (n=4 animals each). S
phase for the last KA' interneurons produced occurs between 10 and 12
hpf. Error bars represent s.e.m. Scale bar: 25 µm.
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