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Vertebrate kidney tubules elongate using a planar cell polarity–dependent, rosette-based mechanism of convergent extension


Cystic kidney diseases are a global public health burden, affecting over 12 million people1. Although much is known about the genetics of kidney development and disease, the cellular mechanisms driving normal kidney tubule elongation remain unclear2,3. Here, we used in vivo imaging to show for the first time that mediolaterally oriented cell intercalation is fundamental to vertebrate kidney morphogenesis. Unexpectedly, we found that kidney tubule elongation is driven in large part by a myosin-dependent, multicellular rosette–based mechanism, previously only described in Drosophila melanogaster. In contrast to findings in Drosophila, however, non-canonical Wnt and planar cell polarity (PCP) signaling is required to control rosette topology and orientation during vertebrate kidney tubule elongation. These data resolve long-standing questions concerning the role of PCP signaling in the developing kidney and, moreover, establish rosette-based intercalation as a deeply conserved cellular engine for epithelial morphogenesis.

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Figure 1: Multicellular rosettes are conserved in mammalian kidney development.
Figure 2: Elongation of vertebrate kidney tubules by mediolateral cell intercalation.
Figure 3: Morphogenetic movements of renal tubule cells employs rosette formation.
Figure 4: Inhibition of myosin interferes with cell movement in Xenopus tubule formation.
Figure 5: PCP signaling controls polarized resolution of multicellular rosettes in developing vertebrate kidney tubules.


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We would like to thank A. Ewald for critical comments on the manuscript; A. Ley for excellent technical assistance, the Life Imaging Center (LIC) at the Centre of Systems Biology at Freiburg University for technical support with confocal microscopy and, in particular, S. Jin form the LIC for programming the tracking macro used in the acquisition of some time-lapse analysis; M. Keuper for digital image analysis; and E.J. Bellefroid (Bruxelles, Belgium) for providing the pCS2+MT-GR plasmid. S.S.L. is supported by the Deutsche Forschungsgemeinschaft (DFG) as part of the Klinische Forschergruppe (KFO) 201. T.J.C. is supported by grants from the March of Dimes, the US National Institutes of Health (NIH; 1R01DK080004) and the UT Southwestern O'Brien Kidney Research Foundation (NIH P30DK079328). O.R., K.L. and G.W. are supported by the Excellence Initiative of the German Federal and State Governments (EXC 294-BIOSS). J.B.W. is supported by the NIH/National Institute of General Medical Sciences (NIGMS), the March of Dimes, the Burroughs Wellcome Fund and the UT Southwestern O'Brien Kidney Research Center (NIH P30DK079328). J.B.W. is an Early Career Scientist of the Howard Hughes Medical Institute. G.W. is supported by the DFG KFO 201 and by the European Community's Seventh Framework Programme (grant agreement 241955, SYSCILIA).

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Authors and Affiliations



S.S.L., C.M.K., T.J.C., J.B.W. and G.W. designed the experiments, S.S.L., K.L., C.M.K. and O.R. performed experiments. S.S.L., K.L., C.M.K., T.J.C., O.R., J.B.W. and G.W. analyzed the data and wrote the manuscript.

Corresponding authors

Correspondence to John B Wallingford or Gerd Walz.

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The authors declare no competing financial interests.

Supplementary information

Supplementary Text and Figures

Supplementary Figures 1–14 and Supplementary Note (PDF 2099 kb)

Supplementary Movie 1

Changes in tubule length and diameter during development. Confocal stacks of staged embryos were acquired after staining for β-Catenin (cell membranes, red), Tomato-Lectin (tubule epithelial membranes, green) and DAPI (nuclei, blue). The epidermis was optically removed. (MOV 4341 kb)

Supplementary Movie 2

Tissue rearrangement during pronephros formation. Confocal time-lapse recordings were performed at low magnification (10 x objective) in 12-minute intervals. Cells were labeled by membrane GFP (green) and nuclear (Histone-)RFP (red) in the pronephros of Xenopus embryos. (MOV 4153 kb)

Supplementary Movie 3

Convergent extension during tubule formation. Xenopus tubule cells were labeled by membrane-associated GFP (gray); time lapse-recordings were performed at higher magnification (25 x objective) in 5-minute intervals. A layer of highest contrast was extracted (see Suppl Note). (MOV 4160 kb)

Supplementary Movie 4

Convergent extension during tubule formation – 2. A filtered image of Suppl Movie 3 was used for cell tracking and segmentation. A subset of cells that is visible throughout the movie was randomly colored to visualize cell rearrangement during tubule extension. (MOV 2924 kb)

Supplementary Movie 5

Rosette formation during tubule elongation. Cell tracking and rosette detection was performed on Suppl Movie 3. Tracked cells are labeled by colored diamonds and rosettes formed by at least 5 cells were circled in yellow. (MOV 8193 kb)

Supplementary Movie 6

Blebbistatin inhibits directed cell movements. Cells were labeled by membrane-associated RFP (gray). Simultaneous time-lapse recordings were performed of DMSO- and blebbistatin-treated Xenopus embryos of the same batch. Cells were tracked by yellow spots. (MOV 2442 kb)

Supplementary Movie 7

formation is reduced in blebbistatin treated embryos. Filtered images of Suppl. Movie 5 superimposed with the result of semi-automated rosette detection (see Supplementary Material and Methods). Rosettes that result from the same cells in multiple frames received the same color and were counted once. (MOV 5459 kb)

Supplementary Movie 8

Xdd1 expression disorients cell movement. The Xdd1-GR expression was activated by dexamethasone treatment in Xenopus embryos. After time lapse recording, cells were tracked by randomly colored spots. (MOV 3789 kb)

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Lienkamp, S., Liu, K., Karner, C. et al. Vertebrate kidney tubules elongate using a planar cell polarity–dependent, rosette-based mechanism of convergent extension. Nat Genet 44, 1382–1387 (2012).

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