Kidney Development

Part I: Organ Culture, Tissue Manipulation, and Live Imaging

1. Dissecting and Culturing and Imaging the Mouse Urogenital System

Paul N. Riccio and Odyssé Michos

2. In Vitro Culture of Embryonic Kidney Rudiments and Isolated Ureteric Buds

Xing Zhang, Kevin T. Bush and Sanjay K. Nigam

3. In vitro Induction of Nephrogenesis in Mouse Metanephric Mesenchyme with Lithium Introduction and Ureteric Bud Recombination

Kimmo Halt and Seppo Vainio

4. Live Imaging of the Developing Mouse Mesonephros

David Grote, Michael Marcotte, and Maxime Bouchard

5. Organotypic Culture of the Urogenital Tract

Ekaterina Batourina, Devangini Gandhi, Cathy L. Mendelsohn and Andrei Molotkov

6. Live Imaging Kidney Development in Zebrafish

Aleksandr Vasilyev and Iain A. Drummond

Part II: Analyzing Ureteric Bud Branching and Nephrogenesis

7. Analysis of 3D Branching Pattern: Hematoxylin and Eosin Method

Sunder Sims-Lucas

8. Three-Dimensional Imaging of Fetal Mouse Kidneys

Deborah Hyink

9. Analysis of Native Kidney Structures in Three Dimensions

Kieran M. Short and Ian M. Smyth

10. Estimating Nephron Number in the Developing Kidney using the Physical Disector/Fractionator Combination

Luise A. Cullen-McEwen, James A. Armitage, Jens R. Nyengaard and John F. Bertram

11. An Immunofluorescence Method to Analyze the Proliferation Status of Individual Nephron Segments in the Xenopus Pronephric Kidney

Daniel Romaker, Bo Zhang and Oliver Wessely

Part III: Renal Primary Cell Cultures and Cell Lines

12. Dissociation of Embryonic Kidney Followed by Re-aggregation as a Method for Chimeric Analysis

Jamie Davies, Mathieu Unbekandt, Jess Ineson, Michael Lusis and Melissa Little

13. Analysis of Migration in Primary Ureteric Bud Epithelial Cells

Satu Kuure

14. Investigating Primary Cilia in Cultured Metanephric Mesenchymal Cells

Lijun Chi and Norman Rosenblum

15. Making Immortalized Cell Lines from Embryonic Mouse Kidney

Guanping Tai, Peter Hohenstein and Jamie Davies

16. Engineered Tissues to Quantify Collective Cell Migration During Morphogenesis

Part IV: Detecting Gene/Protein Expression and Signaling

17. Access and use of the GUDMAP Database of Genitourinary Development

Jamie A. Davies, Melissa H. Little, Bruce Aronow, Jane Armstrong, Jane Brennan, Sue Lloyd-MacGilp, Chris Armit, Simon Harding, Xinjun Piu, Yogmatee Roochun, Bernard Haggarty, Derek Houghton, Duncan Davidson and Richard Baldock

18. Isolation of High Quality RNA from Embryonic Kidney and Cells

Shifaan Thowfeequ and Odyssé Michos

19. Laser Capture

Steven S. Potter and Eric W. Brunskill

20. Use of in Situ Hybridization to Examine Gene Expression in the Embryonic, Neonatal and Adult Urogenital System

Bree A. Rumballe, Han Sheng Chiu, Kylie M. Georgas and Melissa H. Little

21. Detection of β-galactosidase Activity: X-gal Staining

Sally F. Burn

22. Fluorescent Immunolabeling of Embryonic Kidney Samples

Cristina Cebrián

23. Immunohistochemical Staining of dpERK Staining During Early Kidney Development

Xuan Chi and Odyssé Michos

24. Sensing BMP Pathway Activity by Immune Detection of Phosphorylated R-Smad Proteins in Mouse Embryonic Kidney

Javier Lopez-Rios

25. Analysis of in vivo Transcription Factor Recruitment by Chromatin Immunoprecipitation (ChIP) of Mouse Embryonic Kidney

Claire Heliot and Silvia Cereghini

Part V: Manipulating Gene Expression in Developing and Adult Kidney

26. siRNA-mediated RNA Interference in Embryonic Kidney Organ Culture

Jamie A. Davies and Mathieu Unbekandt

27. Morpholino-mediated Gene Knockdown in Mammalian Organ Culture

Alda