This second edition expands upon the previous volume with additional emphasis on recent innovation in basic renal research and has a more holistic approach on associated disorders such as complications associated with peritoneal dialysis, ischemic acute kidney injury, the sympathetic nervous system, and vascular calcification. The book is divided into five parts: Part I provides a number of in vitro, in vivo, and ex vivo models of kidney disease and associated complications; Part II looks at recent advances in imaging techniques; Part III covers recent developments in studying metabolism in renal ischemia and reperfusion; Part IV addresses study and measurement of vascular calcification; and Part V explores analytical techniques that are both topical and of widespread relevance to the study of experimental renal 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 tips on troubleshooting and avoiding known pitfalls. Authoritative and cutting-edge, Kidney Research: Experimental Protocols, Second Edition is a valuable collection of protocols useful to new and experienced researchers who are interested in the field of Nephrology.

Somuchofwhatweknowaboutthepathogenesisofhumandiseasehascomefromthe systematic and careful study of histological material. Indeed, every internal medicine discipline has its landmark papers describing the clinico-pathological correlations. However, increasingly, it is molecular and cellular biology that provides the necessary mechanistic insights. For many years, it was thought that the two skill sets were mutually exclusive, but we hope that this book shows that this is not necessarily so. Implicitinthescienceofhistologyisthepreservationandarchivingoftissue.PartIof the book concentrates on the preparation of tissue, providing an overview of fixation, embedding, and processing (Chapter 1), and in Chapters 2 and 3, the required techniques for the retrieval of RNA from histological sections. Both routine and specialist histological staining techniques are provided in Part II. These include pro- cols for immuno (Chapters 4-7), lectin (Chapter 8), and hybridization (Chapter 9) histochemistry, histologicalstaining (Chapters10and11), aswellasspecificmethods for the in situ identification of hypoxia (Chapter 12) and apoptosis (Chapter 13). Finally, Part III details advances in imaging (Chapters 14-16) and image analysis (Chapter 17). It is hoped that this volume will provide molecular biologists with the basic his- chemical techniques and histologists with the molecular techniques to realise the potential of their resource. We are indebted to the authors for their generosity in sharing these protocol

This third edition provides updated and new chapters on recent innovations in basic renal research. Chapters are divided into three sections covering in vitro, in vivo, and ex vivo models of kidney disease, recent advances in imaging techniques and protocols for performing analytical and functional measurements in the kidney; techniques that are both topical and of widespread relevance to the study of kidney biology and disease. Written in the format of the highly successful Methods in Molecular Biology series, each chapter includes an introduction to the topic, lists necessary materials and methods, includes tips on troubleshooting and known pitfalls, and step-by-step, readily reproducible protocols.   Authoritative and cutting-edge, Kidney Research: Experimental Protocols, Third Edition aims to be comprehensive guide for researchers in the field.

The technique of in situ hybridization, in its various forms, has been used routinely in many laboratories for a number of years. In the post-genome era, gene arrays and proteomics have allowed us to identify hitherto unknown unrecognized pathways and mechanisms. However, rather than diminish the importance of in situ hybridization, the now widespread use of screening te- nologies has increased the need to temporally and spatially localize the dist- bution of mRNA expression. Our intention, in In Situ Hybridization Protocols is to provide ample inf- mation for novices planning to set up the in situ hybridization technique and use it in their laboratory for the first time, as well as giving updates of recent developments for those laboratories where in situ hybridization techniques are already in use. Despite its widespread significance, in situ hybridization has retained a re- tation as one of the more difficult and capricious molecular biological te- niques. This may in part be because of the hybrid nature of the technique, which often requires a mixture of molecular biological and histological skills. The two techniques are usually taught and acquired in different streams of biolo- cal science. The step-by-step and detailed protocols provided in In Situ Hybridization Protocols by researchers active in the field should make it p- sible for both the molecular biologist with little experience of histology and the histologist with little experience of molecular biology to use the technique s- cessfully in their laboratories.

This second edition expands upon the previous volume with additional emphasis on recent innovation in basic renal research and has a more holistic approach on associated disorders such as complications associated with peritoneal dialysis, ischemic acute kidney injury, the sympathetic nervous system, and vascular calcification. The book is divided into five parts: Part I provides a number of in vitro, in vivo, and ex vivo models of kidney disease and associated complications; Part II looks at recent advances in imaging techniques; Part III covers recent developments in studying metabolism in renal ischemia and reperfusion; Part IV addresses study and measurement of vascular calcification; and Part V explores analytical techniques that are both topical and of widespread relevance to the study of experimental renal 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 tips on troubleshooting and avoiding known pitfalls. Authoritative and cutting-edge, Kidney Research: Experimental Protocols, Second Edition is a valuable collection of protocols useful to new and experienced researchers who are interested in the field of Nephrology.

As the expense of treating a growing number of end-stage kidney disease patients increases, greater attention has been paid to prevention and early treatment. The study of renal disease, however, suffers due to the complex nature of renal anatomy and physiology and the plethora of different cell types found in the kidney. In Kidney Research: Experimental Protocols, top experts in the field seek to aid researchers by providing a number of specialized techniques developed to examine this intricate system. Through both well-established and novel methods, this volume explores the preparation and culture of the main cell types used to study renal disease mechanisms, the common animal models used to mimic the various forms of human renal disease, and specific applications and techniques used in vivo and in vitro. Written in the highly successful Methods in Molecular Biology (TM) series format, the chapters contain step-by-step, readily reproducible laboratory protocols, lists of the necessary materials and reagents, and tips on troubleshooting and avoiding known pitfalls. Essential and cutting-edge, Kidney Research: Experimental Protocols delivers invaluable guidance to new and experienced laboratory researchers delving into the sophisticated study of the kidney.

Somuchofwhatweknowaboutthepathogenesisofhumandiseasehascomefromthe systematic and careful study of histological material. Indeed, every internal medicine discipline has its landmark papers describing the clinico-pathological correlations. However, increasingly, it is molecular and cellular biology that provides the necessary mechanistic insights. For many years, it was thought that the two skill sets were mutually exclusive, but we hope that this book shows that this is not necessarily so. Implicitinthescienceofhistologyisthepreservationandarchivingoftissue.PartIof the book concentrates on the preparation of tissue, providing an overview of fixation, embedding, and processing (Chapter 1), and in Chapters 2 and 3, the required techniques for the retrieval of RNA from histological sections. Both routine and specialist histological staining techniques are provided in Part II. These include pro- cols for immuno (Chapters 4-7), lectin (Chapter 8), and hybridization (Chapter 9) histochemistry,histologicalstaining (Chapters10and11),aswellasspecificmethods for the in situ identification of hypoxia (Chapter 12) and apoptosis (Chapter 13). Finally, Part III details advances in imaging (Chapters 14-16) and image analysis (Chapter 17). It is hoped that this volume will provide molecular biologists with the basic his- chemical techniques and histologists with the molecular techniques to realise the potential of their resource. We are indebted to the authors for their generosity in sharing these protocols.

The technique of in situ hybridization, in its various forms, has been used routinely in many laboratories for a number of years. In the post-genome era, gene arrays and proteomics have allowed us to identify hitherto unknown unrecognized pathways and mechanisms. However, rather than diminish the importance of in situ hybridization, the now widespread use of screening te- nologies has increased the need to temporally and spatially localize the dist- bution of mRNA expression. Our intention, in In Situ Hybridization Protocols is to provide ample inf- mation for novices planning to set up the in situ hybridization technique and use it in their laboratory for the first time, as well as giving updates of recent developments for those laboratories where in situ hybridization techniques are already in use. Despite its widespread significance, in situ hybridization has retained a re- tation as one of the more difficult and capricious molecular biological te- niques. This may in part be because of the hybrid nature of the technique, which often requires a mixture of molecular biological and histological skills. The two techniques are usually taught and acquired in different streams of biolo- cal science. The step-by-step and detailed protocols provided in In Situ Hybridization Protocols by researchers active in the field should make it p- sible for both the molecular biologist with little experience of histology and the histologist with little experience of molecular biology to use the technique s- cessfully in their laboratories.