ISBN-13: 9781493962556 / Angielski / Miękka / 2016 / 415 str.
ISBN-13: 9781493962556 / Angielski / Miękka / 2016 / 415 str.
Reflecting recent revolutionary advances in our understanding of renal development, this volume details step-by-step protocols researchers can deploy in their own work, including 3-D morphogenetic mapping and assessing kidney malformation and morbidity.
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
Sriram Manivannan, Jason P. Gleghorn and Celeste M. Nelson
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
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.
1997-2024 DolnySlask.com Agencja Internetowa