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Fig. 2. Laser ablation of DFCs alters LR patterning without affecting development
of the midline. Unablated control embryos expressed ntl in DFCs
(arrow) at 60% epiboly (A) and in the notochord at 24 hours post-fertilization
(E). Laser ablation eliminated DFCs (arrow, dorsal view at 60% epiboly) but
did not alter ntl expression in equatorial mesoderm (B) or in
notochord and tailbud at 24 hpf (F). Control embryos (C) and DFC-ablated
embryos (D) were immunostained with anti-acetylated Tubulin antibodies (green)
to detect cilia and anti-Ntl antibodies (red) to detect the notochord (n) and
Kupffers vesicle (KV). Both control and DFC-ablated embryos showed contiguous
Ntl staining in the notochord, even in embryos (n=2/6) that failed to
form KV (D). (G-J) In control embryos, we observed normal expression of
lft1 (arrow) in the left dorsal diencephelon (G) and lft2
(arrow) in the left heart primordia (I). In DFC-ablated embryos, lft1
(arrow) in the diencephalon (H) and lft2 (arrow) in the heart
primordia (J) were frequently reversed. (K-M) Analysis of lft1
expression in the diencephalon (K) and lft2 expression in the heart
primordia (L) in control (n=71) and DFC-ablated (n=17)
embryos. (M) Analysis of lft1 expression plotted against
lft2 expression in DFC-ablated embryos indicated that the predominant
class of DFC-ablated embryos displayed reversal of both brain and heart
markers (yellow bar). L, left; R, right; B, bilateral; A, absent gene
expression.
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