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Imaging Tools for Analysis of the Ureteric Tree in the Developing Mouse Kidney

  • Luise A. Cullen-McEwen
  • Richard J. Young
  • Gabriel Fricout
  • Dominique Jeulin
  • Ian S. Harper
  • Frank Costantini
  • John F. Bertram
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1075)

Abstract

The structure of the ureteric tree in developing mouse and rat kidneys has previously been quantified in two dimensions. While this type of analysis may provide evidence of changes in ureteric growth, these measurements are effectively inaccurate, as the ureteric tree is a three-dimensional (3D) object. Here we describe a method for measuring the ureteric tree in three dimensions. This technique involves (1) culture of the metanephric kidney at embryonic day 12 (mouse) or 14 (rat), (2) whole-mount immunofluorescence to selectively stain ureteric tree epithelium, (3) confocal microscopy to obtain a complete Z series through the ureteric tree, and (4) image analysis algorithms to binarize, skeletonize, and measure individual branch lengths in 3D. This method has been extended to analysis of the same ureteric tree over time (4D). The results obtained provide accurate and precise quantitation of ureteric tree growth in the developing mouse or rat kidney.

Key words

Kidney Confocal Metanephros Development Ureteric tree Three-dimensional Measurement Image processing 

References

  1. 1.
    Clark AT, Bertram JF (1999) Molecular regulation of nephron endowment. Am J Physiol 276(4Pt2):F485–F497PubMedGoogle Scholar
  2. 2.
    Clark AT, Bertram JF (2000) Advances in renal development. Curr Opin Nephrol Hypertens 9(3):247–251PubMedCrossRefGoogle Scholar
  3. 3.
    Burrow CR (2000) Regulatory molecules in kidney development. Pediatr Nephrol 14(3):240–253PubMedCrossRefGoogle Scholar
  4. 4.
    Kuure S, Vuolteenaho R, Vainio S (2000) Kidney morphogenesis: cellular and molecular regulation. Mech Dev 92(1):31–45PubMedCrossRefGoogle Scholar
  5. 5.
    Pohl M, Stuart RO, Sakurai H, Nigam SK (2000) Branching morphogenesis during kidney development. Annu Rev Physiol 62:595–620PubMedCrossRefGoogle Scholar
  6. 6.
    Davies JA, Fischer CE (2002) Genes and proteins in renal development. Exp Nephrol 10(2):102–113PubMedCrossRefGoogle Scholar
  7. 7.
    Bard JB (2002) Growth and death in the developing mammalian kidney: signals, receptors and conversations. Bioessays 24(1):72–82PubMedCrossRefGoogle Scholar
  8. 8.
    Sakurai H (2003) Molecular mechanism of ureteric bud development. Semin Cell Dev Biol 14:217–224PubMedCrossRefGoogle Scholar
  9. 9.
    Cain JE, Nion T, Jeulin D, Bertram JF (2005) Exogenous BMP-4 amplifies asymmetric ureteric branching in the developing mouse kidney in vitro. Kidney Int 67:420–431PubMedCrossRefGoogle Scholar
  10. 10.
    Cullen-McEwen LA, Fricout G, Harper IS, Jeulin D, Bertram JF (2002) Quantitation of 3-dimensional ureteric branching morphogenesis in cultured embryonic mouse kidney. Int J Dev Biol 46(8):1049–1055PubMedGoogle Scholar
  11. 11.
    Cain JE, Bertram JF (2006) Ureteric branching morphogenesis in BMP4 heterozygous mutant mice. J Anat 209:745–755PubMedCrossRefGoogle Scholar
  12. 12.
    Srinivas S, Goldberg MR, Watanabe T, D’Agati V, al-Awqati Q, Costantini F (1999) Expression of green fluorescent protein in the ureteric bud of transgenic mice: a new tool for the analysis of ureteric bud morphogenesis. Dev Genet 24(3–4):241–251PubMedCrossRefGoogle Scholar
  13. 13.
    Fricout G, Jeulin D, Cullen McEwen L, Harper IS, Bertram JF (2002) 3-D skeletonization of ureteric trees in developing kidneys. In: Talbot H, Beare R (eds) Mathematical morphology, proceedings of the VIth international symposium-ISMM 2002, Sydney, 3–5 April 2002, CSIRO Publishing, pp 157–164Google Scholar

Copyright information

© Springer Science+Business Media New York 2014

Authors and Affiliations

  • Luise A. Cullen-McEwen
    • 1
  • Richard J. Young
    • 1
  • Gabriel Fricout
    • 2
  • Dominique Jeulin
    • 2
  • Ian S. Harper
    • 1
  • Frank Costantini
    • 3
  • John F. Bertram
    • 1
  1. 1.Department of Anatomy and Developmental BiologyMonash UniversityClaytonAustralia
  2. 2.Centre de Morphologie MathématiqueEcole des Mines de ParisParisFrance
  3. 3.Department of Genetics and DevelopmentColumbia UniversityNew YorkUSA

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