Abstract

Loss of kidney function underlies many renal diseases1. Mammals can partly repair their nephrons (the functional units of the kidney), but cannot form new ones2,3. By contrast, fish add nephrons throughout their lifespan and regenerate nephrons de novo after injury4,5, providing a model for understanding how mammalian renal regeneration may be therapeutically activated. Here we trace the source of new nephrons in the adult zebrafish to small cellular aggregates containing nephron progenitors. Transplantation of single aggregates comprising 10–30 cells is sufficient to engraft adults and generate multiple nephrons. Serial transplantation experiments to test self-renewal revealed that nephron progenitors are long-lived and possess significant replicative potential, consistent with stem-cell activity. Transplantation of mixed nephron progenitors tagged with either green or red fluorescent proteins yielded some mosaic nephrons, indicating that multiple nephron progenitors contribute to a single nephron. Consistent with this, live imaging of nephron formation in transparent larvae showed that nephrogenic aggregates form by the coalescence of multiple cells and then differentiate into nephrons. Taken together, these data demonstrate that the zebrafish kidney probably contains self-renewing nephron stem/progenitor cells. The identification of these cells paves the way to isolating or engineering the equivalent cells in mammals and developing novel renal regenerative therapies.

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Acknowledgements

We thank E. C. Liao for help with suturing, and R. Ethier and L. Gyr for zebrafish care. A.J.D. was supported by the Harvard Stem Cell Institute, the American Society of Nephrology and the National Institutes of Health/National Institute of Diabetes and Digestive and Kidney Diseases (P50DK074030).

Author information

Author notes

    • Alan J. Davidson

    Present address: Department of Molecular Medicine and Pathology, School of Medical Sciences, The University of Auckland, Auckland 1142, New Zealand.

Affiliations

  1. Center for Regenerative Medicine, Massachusetts General Hospital, Boston, Massachusetts 02114, USA

    • Cuong Q. Diep
    • , Rahul C. Deo
    • , Teresa M. Holm
    • , Richard W. Naylor
    • , Natasha Arora
    • , Rebecca A. Wingert
    • , Gordana Djordjevic
    • , Benjamin Lichman
    • , Chad A. Cowan
    •  & Alan J. Davidson
  2. Harvard Medical School, Boston, Massachusetts 02115, USA

    • Cuong Q. Diep
    • , Dongdong Ma
    • , Rahul C. Deo
    • , Teresa M. Holm
    • , Richard W. Naylor
    • , Rebecca A. Wingert
    • , Chad A. Cowan
    • , Robert I. Handin
    •  & Alan J. Davidson
  3. Hematology Division, Brigham and Women’s Hospital, Boston, Massachusetts 02115, USA

    • Dongdong Ma
    •  & Robert I. Handin
  4. Harvard Stem Cell Institute, Cambridge, Massachusetts 02138, USA

    • Rebecca A. Wingert
    • , Chad A. Cowan
    • , Robert I. Handin
    •  & Alan J. Davidson
  5. Leibniz Institute for Age Research, Fritz Lipmann Institute, Jena D-07745, Germany

    • Frank Bollig
    •  & Christoph Englert
  6. Dana-Farber Cancer Institute, Boston, Massachusetts 02115, USA

    • Hao Zhu
  7. Section on Model Synaptic Systems, Laboratory of Molecular Physiology, National Institutes of Health/National Institute on Alcohol Abuse and Alcoholism, Bethesda, Maryland 20892, USA

    • Takanori Ikenaga
    •  & Fumihito Ono
  8. Friedrich-Schiller-University, Jena D-07743, Germany

    • Christoph Englert
  9. Department of Stem Cell and Regenerative Biology, Harvard University, Cambridge, Massachusetts 02138, USA

    • Chad A. Cowan
  10. Department of Developmental Biology, University of Pittsburgh, School of Medicine, Pittsburgh, Pennsylvania 15260, USA

    • Neil A. Hukriede

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Contributions

C.Q.D. and A.J.D. designed the experimental strategy, analysed data, prepared the manuscript, and generated and characterized the Tg(cdh17:EGFP), Tg(cdh17:mCherry) and Tg(wt1b:mCherry) lines. C.Q.D. performed the regeneration, transplants, time course and ablation experiments. C.Q.D., D.M. and R.I.H. made the initial observation that nephron progenitors can be transplanted. N.A.H. generated the Tg(lhx1a:EGFP) line (R01DK069403), F.B. and C.E. generated the Tg(wt1b:EGFP) line, and T.I. and F.O. provided the Tg(pax8:DsRed) line. N.A., R.A.W., G.D. and B.L. analysed kidney expression. H.Z. provided sections of regenerating kidneys. R.C.D., T.M.H., R.W.N., and C.A.C. performed quantitative PCR and microarray analyses. All authors commented on the manuscript.

Competing interests

The authors declare no competing financial interests.

Corresponding author

Correspondence to Alan J. Davidson.

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    Supplementary Information

    This file contains Supplementary Figures 1-8 with legends and Supplementary Table 1- 4.

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DOI

https://doi.org/10.1038/nature09669

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