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Fig. 5. Essential neurogenic function of Mash1 at E11.5 and subsequent
stages in the dorsal spinal cord. (A-D) Spinal cords of
Mash1–/– and control embryos were analyzed by
immunohistochemistry at E12.5. (A,B) Differentiated neurons were visualized
using the TuJ1 antibody (green), and YO-PRO1 (red) was used to stain nuclei.
The width of the progenitor domain is indicated. (C,D) In control embryos,
Lbx1+ cells (green) are rarely found in the progenitor domain, marked by the
expression of Pax7 (red). In Mash1–/– embryos,
many Lbx1+ cells can be found in the progenitor domain co-expressing Lbx1 and
Pax7 (yellow). (E) Numbers of Pax7+ cells in the dorsal progenitor
domain were determined in Mash1–/– (white
columns) and control (black columns) embryos at the indicated time points. The
number of progenitor cells is increased in
Mash1–/– at E11.5 and at subsequent stages.
(F) Quantification of newborn neurons at various developmental stages;
to achieve this, BrdU was injected at various time points, and the number of
dorsal BrdU+/NeuN+ neurons in Mash–/– and
control embryos was determined 24 hours later. There is a decrease in the
number of newborn neurons at E11.5 and at subsequent stages in
Mash1–/– embryos. (G) The proliferation
index of progenitor cells (number of dorsal BrdU+ progenitor cells/number of
Pax7+ progenitor cells after a 2-hour BrdU pulse) was determined in
Mash1–/– and control embryos at the indicated
time points. The proliferation index of progenitor cells is not altered in
Mash1–/– embryos. (H) The
differentiation index (number of BrdU+NeuN+ dorsal neurons/total number of
BrdU+ dorsal cells after a 24 hours BrdU pulse) was determined in
Mash1–/– (white columns),
Mash1Ngn2/Mash1Ngn2 (grey columns) and control
(black columns) embryos at E12.5. The thickness of the optical section shown
in A and B is 10 µm; the thickness of the optical sections shown in C and D
is 1.2 µm. The error bars in E-H represent s.d. Scale bars: 100 µm in
A,B; 50 µm in C,D.
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