Article

Synthetic lateral inhibition governs cell-type bifurcation with robust ratios

  • Nature Communications 6, Article number: 6195 (2015)
  • doi:10.1038/ncomms7195
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Abstract

Cell-type diversity in multicellular organisms is created through a series of binary cell fate decisions. Lateral inhibition controlled by Delta–Notch signalling is the core mechanism for the choice of alternative cell types by homogeneous neighbouring cells. Here, we show that cells engineered with a Delta–Notch-dependent lateral inhibition circuit spontaneously bifurcate into Delta-positive and Notch-active cell populations. The synthetic lateral inhibition circuit comprises transcriptional repression of Delta and intracellular feedback of Lunatic fringe (Lfng). The Lfng-feedback subcircuit, even alone, causes the autonomous cell-type bifurcation. Furthermore, the ratio of two cell populations bifurcated by lateral inhibition is reproducible and robust against perturbation. The cell-type ratio is adjustable by the architecture of the lateral inhibition circuit as well as the degree of cell–cell attachment. Thus, the minimum lateral inhibition mechanism between adjacent cells not only serves as a binary cell-type switch of individual cells but also governs the cell-type ratio at the cell-population level.

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Acknowledgements

We thank J. Miyazaki for the PCAG construct. We also thank I. Wada, T. Shibata, Y. Mukumoto and T. Abe for technical advice, Y. Sakaide for technical assistance and S. Hayashi for critical reading of the manuscript. This work was supported by the following grants to M.E.: the funding from Precursory Research for Embryonic Science and Technology (PRESTO), the Funding Program for Next Generation World-Leading Researchers (NEXT Program), the Special Coordination Fund for Promoting Science and Technology and Grant-in-Aid for Scientific Research on Innovative Areas from Japan Ministry of Education, Culture, Sports, Science and Technology (MEXT).

Author information

Author notes

    • Mitsuhiro Matsuda
    •  & Miki Ebisuya

    Present address: Laboratory for Reconstitutive Developmental Biology, RIKEN Quantitative Biology Center, 2-2-3 Minatojima-minamimachi, Kobe, Hyogo 650-0047, Japan

Affiliations

  1. RIKEN Center for Developmental Biology, Kobe 650-0047, Japan

    • Mitsuhiro Matsuda
    • , Makito Koga
    •  & Miki Ebisuya
  2. Career-Path Promotion Unit for Young Life Scientists, Kyoto University, Kyoto 606-8501, Japan

    • Mitsuhiro Matsuda
    • , Makito Koga
    •  & Miki Ebisuya
  3. Graduate School of Biostudies, Kyoto University, Kyoto 606-8502, Japan

    • Mitsuhiro Matsuda
    • , Makito Koga
    •  & Eisuke Nishida
  4. Center for iPS Cell Research and Application, Kyoto University, Kyoto 606-8507, Japan

    • Knut Woltjen
  5. Hakubi Center for Advanced Research, Kyoto University, Kyoto 606-8501, Japan

    • Knut Woltjen
  6. Precursory Research for Embryonic Science and Technology (PRESTO), Japan Science and Technology Agency (JST), Saitama 332-0012, Japan

    • Miki Ebisuya

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Contributions

M.M. and M.E. designed the study. M.M., M.K. and M.E. performed the experiments and analysed the data. K.W. provided the piggyBac system. M.M., E.N. and M.E. wrote the manuscript. All authors discussed the results and commented on the manuscript.

Competing interests

The authors declare no competing financial interests.

Corresponding author

Correspondence to Miki Ebisuya.

Supplementary information

PDF files

  1. 1.

    Supplementary Information

    Supplementary Figures 1-15.

Videos

  1. 1.

    Supplementary Movie 1

    Generation of a salt-and-pepper pattern of red cells and green cells Left: phase, Right: fluorescence. Interval = 1 hour, total time = 205 hours.

  2. 2.

    Supplementary Movie 2

    Cell-type bifurcation in sorted cell populations Left: sorted red cell population, Right: sorted green cell population. Interval = 1 hour, total time = 73 hours.

  3. 3.

    Supplementary Movie 3

    Dynamics of cell state transitions Left: DAPT washout, Right: w/o DAPT (steady state). Arrowheads indicate transitions. Interval = 15 min, total time = 74 hours 45 min.

  4. 4.

    Supplementary Movie 4

    Cell-type bifurcation between two daughter cells Montage of 3 fields. Interval = 0.5 hour, total time = 52 hours.

  5. 5.

    Supplementary Movie 5

    Clustering of N-cadherin-expressing cells Left: control CHO cells, Right: N-cadherin-expressing cells. Interval = 15 min, total time = 20 hours.

Excel files

  1. 1.

    Supplementary Dataset 1

    Model parameters

  2. 2.

    Supplementary Dataset 2

    List of genetic constructs

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