Figure 5

Cardiac neural crest cells are ciliated during their migration and initial migration is normal in cbs/cbs mutants. (A-J) An antibody recognizing p75 was used to label migratory cardiac NCC in transverse sections of E10.5 wild type (+/+, A,B,E-G) and cbs/cbs (C,D,H-J) embryos. (A,B,C,D,F,G,I,J) In each paired panel (for example, A,B) a four-channel labeling is split into two panels for ease of interpretation. (A,C) Initial migration of cohorts of cardiac NCC lateral to the dorsal caudal hindbrain is seen in both wild type and cbs/cbs embryos. (B,D) Colabeling of the cells seen in (A,B) with antibodies recognizing Ift88 (red) to label the ciliary axoneme and anti-γ tubulin (white) to label the basal body demonstrate cilia born by p75-positive cells in both wild type and cbs/cbs embryos. (A-D) Arrowheads mark cilia in (B,D) and are superimposed upon the p75/DAPI stain in (A,C). Dorsal is to the left, lateral to the top. (E,H) Arrowheads indicate p75-positive (green) cardiac NCC migrating along the third pharyngeal artery in both wild type and cbs/cbs embryos. ‘*’: The lumen of the the pharyngeal artery with nucleated red blood cells. (F,G,I,J) High magnification view of migrating cardiac NCC in the same area shown in (E,H). (G,J) Colabeling of the cells seen in (F,I) with antibodies recognizing Ift88 (red) to label the ciliary axoneme and anti-γ tubulin (white) to label the basal body demonstrate cilia born by p75-positive cells in wild type embryos but isolated, non-cilium-associated basal bodies in cbs/cbs embryos. (F,G,I,J) Arrowheads mark cilia in (G,J) and are superimposed upon the p75/DAPI stain in (F,I). (E-J) Dorsal is to the top, lateral to the right. (A,C,E,F,H,I) Sections were stained with DAPI to label nuclei. Scale bars: A-D,F,G,I,J, 10 μm; E,H, 100 μm. cbs, cobblestone; DAPI, 4’-6-diamidino-2-phenylindole; E10.5,embryonic day 10.5; NCC, neural crest cells.