Article | Published:

Lineage specificity of primary cilia in the mouse embryo

Nature Cell Biology volume 17, pages 113122 (2015) | Download Citation

Abstract

Primary cilia are required for vertebrate cells to respond to specific intercellular signals. Here we define when and where primary cilia appear in the mouse embryo using a transgenic line that expresses ARL13B–mCherry in cilia and Centrin 2–GFP in centrosomes. Primary cilia first appear on cells of the epiblast at E6.0 and are subsequently present on all derivatives of the epiblast. In contrast, extraembryonic cells of the visceral endoderm and trophectoderm lineages have centrosomes but no cilia. Stem cell lines derived from embryonic lineages recapitulate the in vivo pattern: epiblast stem cells are ciliated, whereas trophoblast stem cells and extraembryonic endoderm (XEN) stem cells lack cilia. Basal bodies in XEN cells are mature and can form cilia when the AURKA–HDAC6 cilium disassembly pathway is inhibited. The lineage-dependent distribution of cilia is stable throughout much of gestation, defining which cells in the placenta and yolk sac are able to respond to Hedgehog ligands.

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Acknowledgements

We thank H. Bazzi and S. Goetz for comments on the manuscript and M. Distinti for help with manuscript preparation. We thank the MSKCC Mouse Genetics Facility for help making the transgenic mice, and the Molecular Cytology Facility and the Rockefeller University Bio-Imaging Resource Center for help with imaging. This work was supported by National Institutes of Health grants NS044385 and HD035455 to K.V.A., a NYSTEM postdoctoral fellowship to F.K.B., NYSTEM grant 029568 to A-K.H. and an MSKCC Cancer Center Support Grant (P30 CA008748).

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Affiliations

  1. Developmental Biology Program, Sloan Kettering Institute, Memorial Sloan Kettering Cancer Center, 1275 York Avenue New York, New York 10065, USA

    • Fiona K. Bangs
    • , Nadine Schrode
    • , Anna-Katerina Hadjantonakis
    •  & Kathryn V. Anderson

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Contributions

F.K.B. and K.V.A. designed the experiments and wrote the article. F.K.B. carried out the experiments. N.S. and A-K.H. provided advice on stem cell derivation and culture. A-K.H. provided reagents and mice.

Competing interests

The authors declare no competing financial interests.

Corresponding author

Correspondence to Kathryn V. Anderson.

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Videos

  1. 1.

    3D reconstruction of confocal imaging of a 32 cell blastocyst expressing ARL13B-mCherry.

    Centrioles marked with Centrin2-GFP (green) are present in ICM and TE cells but primary cilia are not present in the embryo at this stage.

  2. 2.

    3D reconstruction of confocal imaging of a 72 cell blastocyst expressing ARL13B-mCherry and Centrin2-GFP.

    On rare occasions (2% of cells), ARL13B-mCherry expression can be detected adjacent to centrosomes in cells of the ICM but not in trophectoderm cells.

  3. 3.

    3D reconstruction of confocal imaging of an e5.5 embryo after cavitation, expressing ARL13B-mCherry and Centrin2-GFP.

    Some ARL13b-mCherry expression can be detected adjacent to centrosomes in the epiblast but not the extraembryonic visceral endoderm or trophoectoderm lineages.

  4. 4.

    3D reconstruction of confocal imaging of an e6.0 embryo expressing ARL13B-mCherry (red) and Centrin2-GFP.

    Primary cilia are only present on epiblast cells and not on cells of the visceral endoderm.

  5. 5.

    3D reconstruction of confocal imaging of the distal region and node of an 8.0 embryo.

    Primary cilia expressing ARL13B-mCherry (red) are broadly distributed in both the long cilia of the node (the pit at the centre of the Video) and in cells of all three germ layers. Centrioles are marked with Centrin2-GFP (green).

  6. 6.

    3D reconstruction of confocal imaging of an early bud embryo (e7.5), AFP-GFP (green) is expressed in visceral endoderm cells.

    ARL13B-mCherry (red) labels primary cilia, which are only present on non-GFP expressing definitive endoderm cells.

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DOI

https://doi.org/10.1038/ncb3091

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