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. 2017 Sep 7;7:10751. doi: 10.1038/s41598-017-10822-z

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© The Author(s) 2017

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Ciliary beating direction and basal structure orientation are initially random and then become aligned. (A) Swimming trajectories of embryos. Ten images acquired at 0.3 second intervals are superimposed. hps, hours post fertilization. Scale, 0.5 mm. (B) Mean swimming velocities of embryos of different ages. N = 45–88 from 3–5 embryos. (C) Quantitative comparison of ciliary beating directions. A, anterior; P, posterior. N = 158 (14 h), 125 (24 h) from 8–9 embryos. *p < 0.001. (D) Schematic of angular analysis plotted in E and F. A vector drawn from the cilia transition zone (green) towards the centriole (magenta) gives the direction of ciliary basal structure (black) with respect to the anterior (A) - posterior (P) embryonic axis. A typical immunofluorescence image is shown. (E,F) Phase contrast (left), immunostaining of ciliary basal structures (middle) and circular histograms (right) in two representative embryos at 14 hpf (E) and 24 hpf (F). Yellow arrows indicate the direction in which ciliary basal structures are extended. Scale, 50 μm (left), 10 μm (middle). Circular histograms show the orientation of ciliary basal structures for 23 (14 hpf) and 22 (24 hpf) cilia. CSD, circular standard deviation. (G) Immunoblot of whole embryo proteins with anti-calaxin antibody.