XB-IMG-123890
Xenbase Image ID: 123890
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Figure 2. FGF signaling controls cilia length and directional fluid flow in Kupffer’s Vesicle(a-b) Confocal images of 10 SS embryos, KV labeled with antibodies against aPKC (red) and acetylated tubulin (green). Control and fgfr1 morphants had similar KV structure, but cilia were shorter in fgfr1 morphants (compare insets in a and b). (c) Cilia lengths were significantly different (p< 2.88e-06) in fgfr1 morphants (688 cilia; 18 embryos) versus Control morphants (437 cilia; 9 embryos). Cilia length was similar in WT uninjected (533 cilia; 10 embryos) and Control morphant (p< 0.93), cilia numbers per KV were similar in Control and fgfr1 morphants (p< 0.26). Cilia length defects in fgfr1 morphants were rescued by Xenopus FGFR1 (xFGFR1) mRNA (p< 4.70e-05; 807 cilia; 21 embryos). Injection of xFGFR1 mRNA alone had no affect on cilia length (p<0.73; 526 cilia, 14 embryos). (d) Embryos treated with SU5402 during shield stage (248 cilia; 12 embryos) had shorter cilia compared to DMSO control embryos (p<3.26e-06; 686 cilia; 15 embryos). (e) Cilia were shorter in transgenic DN-FGFR embryos that were heat shocked at 60% epiboly (656 cilia; 19 embryos) compared to heat shocked non-transgenic siblings (p<6.94e-03; 375 cilia; 10 embryos) and non-heat-shocked siblings (p<6.99e-03; 910 cilia; 16 embryos). (f) There was no difference in cilia length (p<0.28) in fgf24 morphants (455 cilia; 10 embryos) versus Control morphants (481 cilia; 10 embryos). However, cilia were shorter when both FGF8 and FGF24 ligands were diminished (fgf24 MO in ace mutants; 12 embryos; 244 cilia), compared to single ligand knockdown (FGF8/ace mutants: p< 1.39e-04; 10 embryos; 480 cilia; fgf24 MO in ace sibs: p< 3.44e-04; 15 embryos; 643 cilia) and WT ace siblings (p<3.63e-07; 13 embryos; 626 cilia). (g-h) DIC images of bead-injected KVs in Control and fgfr1 morphants injected with fluorescent beads. (i-j) Bead paths tracked by Metamorph software. Directional KV fluid flow was absent in fgfr1 morphants (j; p< 6.4e-15; 44 beads, 9 embryos) compared to counterclockwise flow in Control morphants (i; 39 beads, 8 embryos). Error bars are standard error of the mean (s.e.m). Image published in: Neugebauer JM et al. (2009) Image downloaded from an Open Access article in PubMed Central. Image reproduced on Xenbase with permission of the publisher and the copyright holder. Larger Image Printer Friendly View |