Diabetic Nephropathy Methods and Protocols

This book provides a toolkit of novel research approaches for investigators to study diabetic nephropathy, including critical experimental models from the fly to the fish, cells in culture, and in vivo mammalian approaches. The collection also explores powerful techniques to image the kidney, such as traditional histological techniques as well as electron, confocal, and two-photon microscopy, pathophysiology of the diabetic kidney, and gene editing and regenerative medicine. Written for the highly successful Methods in Molecular Biology series, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls. 

Part I: Introduction

 

1. Diabetic Nephropathy: An Overview

            Manpreet K. Sagoo and Luigi Gnudi

 

Part II: Experimental Models to Study Diabetic Nephropathy

 

2. Modeling Podocyte Biology Using Drosophila Nephrocytes

            Paul S. Hartley and Richard J. Coward

 

3. A Technique for Studying Glomerular Filtration Integrity in the Zebrafish Pronephros

            Maria Kolatsi-Joannou and Daniel Osborn

 

4. Modeling Human Diabetic Kidney Disease by Combining Hyperglycemia and Hypertension in a Transgenic Rodent Model

            Carolynn Cairns and Bryan Conway

 

5. Isolating and Culturing Mouse Podocyte Cells to Study Diabetic Nephropathy

            Elisavet Vasilopoulou

 

Part III: Techniques to Image the Diabetic Kidney

 

6. Histological Examination of the Diabetic Kidney

            Camillo Carrara, Mauro Abbate, Sara Conti, Daniela Rottoli, Paola Rizzo, and Gianfranco Marchetti

 

7. Sample Preparation and Stereological Methods for the Study of Glomerular Ultrastructure Using Electron Microscopy

            Kathryn E. White

 

8. Tissue Clearing and Deep Imaging of the Kidney Using Confocal and Two-Photon Microscopy

            Daniyal J. Jafree, David A. Long, Peter J. Scambler, and Dale Moulding

 

Part IV: Assessing the Function and Pathophysiological Properties of the Diabetic Kidney

 

9. Transcutaneous Measurement of Glomerular Filtration Rate in Rodents

            Cristina Daniele, Daniela Nardozi, Angelo Torelli, Arif ul Maula Khan, and Norbert Gretz

 

10. An In Vitro Method to Analyze Glucose Uptake in Podocytes

            Abigail C. Lay and Richard J. Coward

 

11. Glomerular Endothelial Cells: Assessment of Barrier Properties In Vitro

            Raina D. Ramnath and Simon C. Satchell

 

12. Methods to Detect Endoplasmic Reticulum Stress and Apoptosis in Diabetic Nephropathy

            Khurrum Shahzad, Sanchita Ghosh, Akash Mathew, and Berend Isermann

 

13. Isolating Urinary Extracellular Vesicles as Biomarkers for Diabetic Disease

            Karina Barreiro, Tobias B. Huber, and Harry Holthofer

 

14. Examining Cell-Cell Interactions in the Kidney Using AFM Single-Cell Force Spectroscopy

            Eleftherios Siamantouras, Claire E. Hills, Kuo-Kang Liu, and Paul E. Squires

 

Part V: Transcriptome, Metabolic, and Proteomic Profiling of the Diabetic Kidney

 

15. Genetic Strategies to Understand Human Diabetic Nephropathy: Wet-Lab Approaches

            Laura J. Smyth, Katie Kerr, Seamus Duffy, Jill Kilner, and Amy Jayne McKnight

 

16. Genetic Strategies to Understand Human Diabetic Nephropathy: In Silico Strategies for Molecular Data - Association Studies

            Marisa Canadas-Garre, Laura J. Smyth, Kerry Anderson, Katie Kerr, and Amy Jayne McKnight

 

17. Assessment of Urinary MicroRNAs by Quantitative Polymerase Chain Reaction in Diabetic Nephropathy Patients

            Lucy Jade Newbury, Alexa Wonnacott, Kate Simpson, Timothy Bowen, and Donald Fraser

 

18. Metabolomic and Proteomic Techniques for Establishing Biomarkers and Improving Our Understanding of Pathophysiology in Diabetic Nephropathy

            Justyna Siwy, Linda Ahonen, Pedro Magalhães, Maria Franzi, and Peter Rossing

 

Part VI: New Approaches: Gene Editing and Stem Cells to Study Diabetic Kidney Disease

 

19. Formation of Mature Nephrons by Implantation of Human Pluripotent Stem Cell-Derived Progenitors into Mice

            Ioannis Bantounas, Edina Silajdžic, Adrian S. Woolf, and Susan J. Kimber

 

20. Generating Mutant Renal Cell Lines Using CRISPR Technologies

            Nuria Perretta Tejedor, Grace M. Freke, Marian Seda, David A. Long, and Dagan Jenkins