Abstract
Accurate analysis of the three-dimensional (3D) architecture of developing organs is critical to understanding how developmental defects can be linked with structural abnormalities. Here, we describe a 3D reconstruction technique of the developing kidney including the outer kidney capsule, ureteric epithelium, and developing nephrons. This 3D reconstructive process involves generating serial sections of the developing kidney, followed by histological staining. Each serial image is projected on the monitor and each tissue lineage or structure is traced. The kidney tracings are aligned and a 3D image is rendered. Each reconstructed tissue/lineage can then be subjected to quantitative analysis (e.g., surface area or volume). The reconstructed ureteric epithelium can be skeletonized to determine the branching architecture.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Saxen L, Sariola H (1987) Early organogenesis of the kidney. Pediatr Nephrol 1(3):385–392
Dressler GR (2006) The cellular basis of kidney development. Annu Rev Cell Dev Biol 22:509–529
Costantini F (2006) Renal branching morphogenesis: concepts, questions, and recent advances. Differentiation 74(7):402–421
Brenner BM, Garcia DL, Anderson S (1988) Glomeruli and blood pressure. Less of one, more the other? Am J Hypertens 1(4 Pt 1):335–347
Douglas-Denton RN, McNamara BJ, Hoy WE, Hughson MD, Bertram JF (2006) Does nephron number matter in the development of kidney disease? Ethn Dis 16(2 Suppl 2):S2-40–S2-45
Clark A, Bertram J (2000) Advances in renal development. Curr Opin Nephrol Hypertens 9(May):247–251
Clark AT, Bertram JF (1999) Molecular regulation of nephron endowment. Am J Physiol 276(4 Pt 2):F485–F497
Davies JA (1996) Mesenchyme to epithelium transition during development of the mammalian kidney tubule. Acta Anat 156(3):187–201
Davies JA, Bard JB (1998) The development of the kidney. Curr Top Dev Biol 39:245–301
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(2):420–431
Cebrian C, Borodo K, Charles N, Herzlinger DA (2004) Morphometric index of the developing murine kidney. Dev Dyn 231(3):601–608
Clendenon JL, Byars JM, Hyink DP (2006) Image processing software for 3D light microscopy. Nephron Exp Nephrol 103(2):e50–e54
Short KM, Hodson MJ, Smyth IM (2010) Tomographic quantification of branching morphogenesis and renal development. Kidney Int 77(12):1132–1139
Sims-Lucas S, Argyropoulos C, Kish K et al (2009) Three-dimensional imaging reveals ureteric and mesenchymal defects in Fgfr2-mutant kidneys. J Am Soc Nephrol 20(12):2525–2533
Sims-Lucas S, Cusack B, Eswarakumar VP et al (2011) Independent roles of Fgfr2 and Frs2{alpha} in ureteric epithelium. Development 138(7):1275–1280
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2012 Springer Science+Business Media, LLC
About this protocol
Cite this protocol
Sims-Lucas, S. (2012). Analysis of 3D Branching Pattern: Hematoxylin and Eosin Method. In: Michos, O. (eds) Kidney Development. Methods in Molecular Biology™, vol 886. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-61779-851-1_7
Download citation
DOI: https://doi.org/10.1007/978-1-61779-851-1_7
Published:
Publisher Name: Humana Press, Totowa, NJ
Print ISBN: 978-1-61779-850-4
Online ISBN: 978-1-61779-851-1
eBook Packages: Springer Protocols