Figure 2. Impaired Ciliation and Leftward Flow upon ATP4 Loss of Function(A�F) Flow analysis. (A) Directionality and (B) velocity of fluorescent beads added to GRP explants at stage 17 were drastically reduced in ATP4a morphants or SCH28080-treated specimens, as compared to wild-type, CoMO-injected, or DMSO-treated embryos. n represents number of explants analyzed. (C�F) Frequency distribution of trajectory angles in representative explants injected with (C) CoMO, (D) ATP4aMO, (E) DMSO, and (F) SCH28080. Dashed circles indicate maximum frequency in histogram specified in percent. a, anterior; l, left; n, number of particles above threshold; p, posterior; r, right; v, average velocity of particles; ρ, quality of flow.(G�K) SEM analysis of GRP ciliation and morphology. (G and H) Representative dorsal explants reveal shorter cilia, fewer ciliated cells, and polarization defects in (H) ATP4a morphant (H) as compared to (G) CoMO-injected specimen. Cell boundaries are indicated by dashed orange lines in higher magnification of SEM pictures in (G′) and (H′). Blowups in (G′) and (H′) illustrate a long and posteriorly polarized cilium in (G′), indicated by a green arrowhead, and two short cilia in (H′), of which one emerges in a central position, indicated by a yellow arrowhead. (G� and H�) Evaluation of cilia polarization. Green, posterior; yellow, other; red, no cilium. (I�K) Quantification of (I) ciliation rate and (J) cilia length of GRP cilia in defined areas, as indicated by white squares in (G) and (H). (K) Cilia polarization was assessed in areas of defined size, as indicated by dashed boxes in (G) and (H). ∗∗Highly significant. ∗∗∗Very highly significant. ns, not significant. Numbers indicate number of dorsal explants (in parentheses) or cilia (in brackets) analyzed.See also Figure S2 and Movie S1.
Image published in: Walentek P et al. (2012)
Copyright © 2012. Image reproduced with permission of the publisher and the copyright holder. This is an Open Access article distributed under the terms of the Creative Commons Attribution License.
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