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Fig. 3. Overexpression of c-Gcm1 promotes premature neuronal
differentiation. Electroporation with control or c-Gcm1
expression vectors in truncal neural tube
(A-B',E-J') or telencephalon (C,D) of
E1.5 embryos. Immunolabeling with ßIII-tubulin (Tuj1 in A-E), HuC/D
(G-H') and Lim1/2 (I-J') neuronal markers were performed 24 hours
after electroporation. (A-H') Electroporation of control vector (green
in A,C,G,I) does not modify the expression pattern of ßIII-tubulin (red
in A,A',C) or HuC/D (red in G,G'), whereas c-Gcm1
overexpression (green in B,D,E,H,J) leads to ectopic differentiation of
Tuj1-positive (red in B,B',D; blue in E, arrows) and HuC/D-positive (red
in H,H') neurons in the neuroepithelium. Inserts in B and H are high
magnifications of c-Gcm1-overexpressing cells that coexpress
ßIII-tubulin and HuC/D, respectively. (E) ßIII-tubulin (blue) and
Pax7 (red) coimmunolabeling shows that c-Gcm1-overexpressing cells
coexpress ßIII-tubulin but not Pax7 (arrows). (I-J') Compared with
control (I,I'), c-Gcm1 overexpression induces the generation of
Lim1/2-positive neurons within the ventricular zone (J,J'). (F)
Percentage of transfected cells expressing ßIII-tubulin (Tuj1) and Lim1/2
after electroporation of control or c-Gcm1 expression vectors.
Differences between the percentage of c-Gcm1
transfected cells expressing neuronal markers versus controls are statistically
significant (asterisks, P<0.001). Scale bars: 40µm in A-D,G-J'; 10µm in
insert in B,H; 20µm in E.
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