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Priming, initiation and synchronization of the segmentation clock by deltaD and deltaC

Nature Cell Biology volume 9, pages 523–530 (2007)Cite this article

Abstract

Zebrafish somitogenesis is governed by a segmentation clock that generates oscillations in expression of several Notch pathway genes, including her1, her7 and deltaC1,2,3,4,5,6. Using a combination of pharmacological inhibition and Mendelian genetics, we show that DeltaD and DeltaC, two Notch ligands, represent functionally distinct signals within the segmentation clock. Using high-resolution fluorescent in situ hybridization7, the oscillations were divided into phases based on eight distinct subcellular patterns of mRNA localization for 140,000 cells. her1, her7 and deltaC expression was examined in wild-type, deltaD−/− and deltaC−/− embryos. We identified areas within the tailbud where the clock is set up in the progenitor cells (priming), where the clock starts running (initiation), and where the clocks of neighbouring cells are entrained (synchronization). We find that the clocks of motile cells are primed by deltaD in a progenitor zone in the posterior tailbud and that deltaD is required for cells to initiate oscillations on exiting this zone. Oscillations of adjacent cells are synchronized and amplified by deltaC in the posterior presomitic mesoderm as cell movement subsides and cells maintain stable neighbour relationships.

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Figure 1: A segmentation-recovery assay.
Figure 2: Eight phases of oscillating gene transcription are defined by subcellular mRNA localization patterns.
Figure 3: Motile somite-precursor cells remain in the progenitor zone for varying amounts of time and do not exhibit oscillation in her1, her7 and deltaC expression until migrating into the initiation zone.
Figure 4: Oscillations are driven by deltaD and synchronized by deltaC.
Figure 5: Oscillation phase distributions are differentially altered in the two delta mutants.

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Acknowledgements

We thank S. Truong and S. Vadasz for contributing the fluorescent in situs and mRNA injections, respectively. We thank R. Ho for providing the sptb16 fish and A. Oates for providing the her7 plasmid. We are grateful to T. Emonet, M. Garcia-Castro, T. Brend and D. Jülich for their critical comments on the manuscript. A.M. was supported by the Department of Homeland Security Scholarship and Fellowship program. This work was funded by a grant from the National Institute of Child Health and Human Development (NICHD; R01 HD045738) to S.A.H.

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Authors and Affiliations

  1. Department of Molecular, Cellular and Developmental Biology, Yale University, New Haven, 06520, CT, USA

    Andrew Mara, Joshua Schroeder & Scott A. Holley

  2. Department of Cell Biology, Yale University School of Medicine, New Haven, 06520, CT, USA

    Cécile Chalouni

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Contributions

A.M. performed all of the experiments, with help from J.S., except for the analysis of cell movement, which was performed by S.A.H. with help from J.S. and C.C. A.M. and S.A.H. conceived and designed the experiments and wrote the manuscript.

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Correspondence to Scott A. Holley.

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The authors declare no competing financial interests.

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Mara, A., Schroeder, J., Chalouni, C. et al. Priming, initiation and synchronization of the segmentation clock by deltaD and deltaC. Nat Cell Biol 9, 523–530 (2007). https://doi.org/10.1038/ncb1578

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