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Identification of adult nephron progenitors capable of kidney regeneration in zebrafish

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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Figure 1: The adult zebrafish kidney undergoes nephrogenesis throughout life and after injury.
Figure 2: The adult zebrafish kidney contains transplantable progenitors that form functional nephrons.
Figure 3: Expression of lhx1a:EGFP and other renal factors in the adult kidney.
Figure 4: lhx1a:EGFP + cells form nephrons during adult kidney development and after transplantation.

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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).

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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.

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Correspondence to Alan J. Davidson.

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The authors declare no competing financial interests.

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Diep, C., Ma, D., Deo, R. et al. Identification of adult nephron progenitors capable of kidney regeneration in zebrafish. Nature 470, 95–100 (2011). https://doi.org/10.1038/nature09669

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