FIGURE

Fig. 2

ID

ZDB-FIG-150617-8

Publication

Yuan et al., 2015 - Intraciliary Calcium Oscillations Initiate Vertebrate Left-Right Asymmetry

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Fig. 2

Cilia-Targeted GECIs Reveal the Presence of ICOs in the LRO of Zebrafish that Are Leftward Biased and Dependent on Pkd2 and Ciliary Motility

(A-F) Spatial mapping of ICOs in the zebrafish LRO.

(A, C, and E) Representative fluorescent live images of the LRO from vehicle-injected (A), pkd2 morphant (MO) (C), and c21orf59 morphant (E) zebrafish embryos at the one-somite stage expressing ratiometric GECI (arl13b-GCaMP6/mCherry) (ratiometric signal is shown and false colored with a rainbow intensometric scale). White arrowheads indicate LRO cilia at resting state, whereas the magenta arrowhead (A) highlights an ICO inducing a cytosolic calcium wave. The scale bars represent 10 µm.

(B, D, and F) Rose diagrams depicting the spatial distribution and mean percentage of cells per embryo displaying ICOs in the total LRO of vehicle (B), pkd2 MO (D), and c21orf59 MO (F) embryos spanning the entire course of LRO development (bud to 16-somite stage). Rings correlate with the mean percentage of cells exhibiting calcium oscillations per embryo from bud to 16-somite stage (n = 10 embryos at all stages for pkd2 and vehicle; n = 5 embryos for c21orf59 and vehicle). Control and experimental samples were acquired in a pairwise manner, and analysis was performed on time-lapse recordings spanning bud to 16-somite stage. Control samples for c21orf59 morphants are not shown, because the distribution and frequency were equivalent to controls for pkd2 morphants. Left-sided calcium, blue; right-sided calcium, orange.

(G) Intraciliary calcium at the LRO exhibits an oscillation-like dynamic. Ratiometric intensity over time plot of a single cilium exhibiting ICOs in vehicle (blue), pkd2 MO (magenta), and c21orf59 MO (green) LROs at the one- to four-somite stage. Each plot was aligned postacquisition to the first detected ICOs (indicated by the black arrowhead) and thresholded (indicated by the black horizontal line) to facilitate analysis of calcium oscillation dynamics.

A, anterior; P, posterior; L, left; R, right. See also Figures S2 and S3 and Movies S1, S2, and S3.

Expression Data

Expression Detail

Antibody Labeling

Phenotype Data

Fish:	WT + MO2-cfap298 WT + MO3-pkd2
Knockdown Reagents:	MO2-cfap298 MO3-pkd2
Observed In:	Kupffer's vesicle calcium-mediated signaling Kupffer's vesicle cilium movement
Stage:	90%-epiboly

Phenotype Detail

Acknowledgments

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