Kidney Development, Softcover reprint of the original 1st ed. 2012 Methods and Protocols Methods in Molecular Biology Series, Vol. 886

Over the last decade the development of new molecular biology tools, advanced microscopy, live imaging and systems biology approaches have revolutionized our conception of how embryonic development proceeds.  One fundamental aspect of development biology is the concept of morphogenesis: understanding how a group of multipotent cells organize and differentiate into a complex organ.  In Kidney Development: Methods and Protocols, expert researchers in the field detail different approaches to tackle kidney development. These approaches include culture and live imaging aspects of kidney development, analyzing the 3-dimensional aspects of branching morphogenesis as well as nephrogenesis, manipulation of the gene/protein expression during kidney development as well as in the adult kidney, and how to assess kidney malformation and disease. Written in the highly successful Methods in Molecular Biology? series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and key tips on troubleshooting and avoiding known pitfalls.

Authoritative and practical, Kidney Development: Methods and Protocols seeks to aid scientists in the further study of the process of morphogenesis which is fundamental important not only for studying developmental biology but also for regenerative medicine.

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 Tufro

28. Microinjection into the Lumen of the Ureteric Tree

Cristina Cebrián

29. Renal Delivery of Adenovirus and Antisense Oligonucleotides in Rats by Retrograde Renal Vein Injection

Guadalupe Ortiz-Muñoz, Beñat Mallavia, Oscar Lopez-Franco, Purificacion Hernandez-Vargas, Jesus Egido and Carmen Gomez-Guerrero

Part VI: Analyzing Functional Defects of the Kidney and Urinary Tract

 30. Estimating Total Nephron Number in the Adult Kidney using the Physical Dissector/Fractionator Combination

Luise A. Cullen-McEwen, Rebecca N. Douglas-Denton and John F Bertram

 31. Assessing Urinary Tract Defects in Mice: Methods to Detect the Presence of Vesico-ureteric Reflux and Urinary Tract Obstruction

Inga J. Murawski Christine L. Watt and Indra R. Gupta

 32. Ischemia-reperfusion Injury of the Mouse Kidney

Leif Oxburgh and Mark P. de Caestecker

 33. Variable Partial Unilateral Ureteral Obstruction and its Release in the Neonatal and Adult Mouse

Barbara A. Thornhill and Robert L. Chevalier

 34. Urinary Diversion Via Cutaneous Vesicostomy in the Megabladder Mouse

Ashley R. Carpenter, Brian Becknell, Daniel A. Hirselj and Kirk M. McHugh

 35. Ultrasound Imaging of the Murine Kidney

Ashley Carpenter, Brian Becknell, Susan E. Ingraham and Kirk M. McHugh

Focuses on mouse models but features other popular models as well

Provides step-by-step detail essential for reproducible results

Contains key notes and implementation advice from the experts

Includes supplementary material: sn.pub/extras