Abstract
It has been proposed that ciliated cells that produce a leftward fluid flow mediate left-right patterning in many vertebrate embryos. The cilia on these cells combine features of primary sensory and motile cilia, but how this cilia subtype is specified is unknown. We address this issue by analyzing the Xenopus and zebrafish homologs of Foxj1, a forkhead transcription factor necessary for ciliogenesis in multiciliated cells of the mouse. We show that the cilia that underlie left-right patterning on the Xenopus gastrocoel roof plate (GRP) and zebrafish Kupffer's vesicle are severely shortened or fail to form in Foxj1 morphants. We also show that misexpressing Foxj1 is sufficient to induce ectopic GRP-like cilia formation in frog embryos. Microarray analysis indicates that Xenopus Foxj1 induces the formation of cilia by upregulating the expression of motile cilia genes. These results indicate that Foxj1 is a critical determinant in the specification of cilia used in left-right patterning.
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Acknowledgements
The authors thank members of the Kintner laboratory for comments on the manuscript, M. Wood for technical assistance with the TEM and B. Mitchell for assistance with high-speed photography in collaboration with C. Yu, P. Taborek and F. Huisman in the Department of Physics at University of California, Irvine. The work was supported by grants from the Kanae Foundation to I.O., from the G. Harold and Leila Y. Mathers and Cellex MEC foundations and US National Institutes of Health to J.C.I.B. and from the US NIH to C.K.
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J.L.S., I.O., J.C.I.B. and C.K. designed the study; J.L.S. and I.O. carried out zebrafish experiments; J.L.S. and C.K. carried out Xenopus experiments; all authors contributed to the interpretation of results and the writing of the manuscript.
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Supplementary Text and Figures
Supplementary Figures 1–9 and Supplementary Tables 1 and 2 (PDF 920 kb)
Supplementary Movie 1
Example of an ectopic cilium induced by Foxj1 with a rotational beat pattern. The movie is 0.035 seconds in duration, and the cilium is beating at 25 Hz. (AVI 2751 kb)
Supplementary Movie 2
Example of an ectopic cilium induced by Foxj1 with a wipe-like beat pattern. The movie is 0.070 seconds in duration, and the cilium is beating at ∼15 Hz. (AVI 1585 kb)
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Stubbs, J., Oishi, I., Izpisúa Belmonte, J. et al. The forkhead protein Foxj1 specifies node-like cilia in Xenopus and zebrafish embryos. Nat Genet 40, 1454–1460 (2008). https://doi.org/10.1038/ng.267
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DOI: https://doi.org/10.1038/ng.267
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