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. 2009 Aug 31;106(37):15774–15779. doi: 10.1073/pnas.0906072106

Fig. 4.

Fig. 4.

Influence of the activity of Bmp antagonists in CNC on Fgf8 expression and brain development. (A) Two-step procedure for CNC transfection. Nucleic acids, injected in the neural groove of quail neurula, are bilaterally transfected into CNC by electroporation before CNC bilateral transplantation into un-transfected chicken embryo. (B) Co-injection and co-electroporation (C) with rhodamine dextran (Invitrogen) enables visualization of the transfected CNC (C; dotted lines) before transplantation. Fgf8 expression at E2 (D-I) and E4 morphology (J-L) of control (D, E, and J), Gremlin Noggin-dsRNA-treated (F, G, and K), and Gremlin Noggin-RCAS-treated (H, I, and L) embryos. Loss of function of Bmp antagonists in CNC alters Fgf8 expression in ANR (F and G; arrowheads). Inversely, Gremlin Noggin over-expression in CNC increases the FgF8 expression in the ANR (H and I; arrowheads) and isthmus, and induces transcript accumulation along the dorsal midline (I; stars). Normal morphology of E4 control embryo (J). (K) The loss of function results in the underdevelopment of cephalic vesicles and eyes. (L) Dorsal expansion of cephalic vesicles resulting from the overexpression of Gremlin and Noggin in CNC (M and N). Whole-mount E8 pre-otic brains: lateral (M) and ventral (N) views show a normal brain (Left) and the hypertrophied morphology (Right) induced by Gremlin Noggin overexpression in CNC.