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Figure 5 | Neural Development

Figure 5

From: A negative modulatory role for rho and rho-associated kinase signaling in delamination of neural crest cells

Figure 5

Inhibition of either RhoA or RhoB activities enhances neural crest (NC) delamination in ovo. (A-C) Transverse sections at a dissociated somite level showing the emigration of control green fluorescent protein (GFP)-expressing (A,A'), N19-RhoB-expressing (B,B') and GAP-RhoB-expressing (C,C') NC cells (green). Note in (A',B',C') that most delaminating progenitors are also bromo-deoxyuridine (BrdU)+ (red; arrows pointing at double-labeled cells). (D,D') Transverse section at a similar level as in (A-C) of a hemi-NT that received N19-RhoB (green) and was further stained for HNK-1 (red in (D)) and FoXD3 mRNA (D' and inset). Transfected emigrating cells are positive both for HNK-1 and FoXD3 (arrows). (E, F) Two transverse sections of embryos that received N19-RhoA (green) and were further processed for HNK-1 immunostaining (E, red) or FoXD3 in situ hybridization (F and inset). Note that delaminating cells that received N19-RhoA also express HNK-1 or FoxD3 (arrows). Note as well that the transfected hemi-NT cells lost their normal pseudostratified appearance and are rounded. (G-I) Transverse sections through the caudal segmental plate (cSP) level (G), rostral segmental plate (rSP) level (H), and recently formed epithelial somite (ES) (I) of embryos that received N19-RhoA. Note premature delamination of NC progenitors expressing N19-rhoA/GFP (arrows) on a background of Hoechst nuclear stain (blue) or co-expressing Snail2 mRNA (inset in (I)). No delamination from these axial levels is observed upon transfection of control GFP (not shown). Note that at these very caudal levels of the axis, electroporation was predominantly dorsal due to positioning of the electrodes at a slightly more rostral level in order not to damage the gastrulating area of the axis. (J) Quantification of NC delamination (*p < 0.05, **p < 0.01, ***p = 0.0001). DS, dissociating somite; ES, epithelial somite; s.d., standard deviation; SP, segmental plate. Bar: 45 μM (A-F); 53 μM (G-I); 83 μM, insets in (D,E,I).

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