VEGF Signaling: Methods and Protocols

ISBN-13: 9781493943708 / Angielski / Miękka / 2016 / 225 str.

Lorna Fiedler

Spis treści

Part I VEGF and VEGF Receptors

1. VEGF Splicing and the Role of VEGF Splice Variants: From Physiological-Pathological Conditions to Specific Pre-mRNA Splicing
Mélanie Guyot and Gilles Pagès

2. Detection and Quantification of VEGF Isoforms by ELISA
Jean-Michel Vernes and Y. Gloria Meng

3. Quantitation of Circulating Neuropilin-1 in Human, Monkey, Mouse, and Rat Sera by ELISA
Yanmei Lu and Y. Gloria Meng

4. Detection and Quantification of Vascular Endothelial Growth Factor Receptor Tyrosine Kinases in Primary Human Endothelial Cells
Gareth W. Fearnley, Stephen B. Wheatcroft, and Sreenivasan Ponnambalam

5. Induction of VEGF Secretion in Cardiomyocytes by Mechanical Stretch
Michelle L. Matter

6. Chromatin Immunoprecipitation Assay: Examining the Interaction of NFkB with the VEGF Promoter
Chad B. Walton and Michelle L. Matter

Part II Modulation and Analysis of VEGF Dependent Signaling

7. An Overview of VEGF-Mediated Signal Transduction
Ian Evans

8. Identification of Receptor Tyrosine Kinase Inhibitors using Cell Surface Biotinylation and Affinity Isolation
Antony M. Latham, Jayakanth Kankanala, Colin W.G. Fishwick and Sreenivasan Ponnambalam

9. Analysis of VEGF-Mediated ERK5 Activity in Endothelial Cells
Gopika N. Nithianandarajah-Jones and Michael J. Cross

10. In Vitro Angiogenesis Assays
Ian Evans

11. Chemotactic Migration of Endothelial Cells Towards VEGF-A165
Caroline Pellet-Many

Part III In Vivo Models to Study VEGF Signaling

12. Vasculogenesis and Angiogenesis in VEGF Receptor-1 Deficient Mice
Vivienne C. Ho and Guo-Hua Fong

13. The Embryonic Mouse Hindbrain and Postnatal Retina as In Vivo Models to Study Angiogenesis
Alessandro Fantin and Christiana Ruhrberg

14. VEGF Gene Transfer to the Utero-Placental Circulation of Pregnant Guinea Pigs to Enhance Fetal Growth
Vedanta Mehta, David J. Carr, Anna Swanson, and Anna L. David

15. VEGF Gene Transfer to the Utero-placental Circulation of Pregnant Sheep to Enhance Fetal Growth
David J. Carr, Vedanta Mehta, Jacqueline M. Wallace, and Anna L. David

16. Generation of Targeted Mutations in Zebrafish Using the CRISPR/Cas System
Linlin Yin, Li-En Jao, and Wenbiao Chen

O autorze

Lorna R. Fielder, Program in cardiovascular and Metabolic Disorders, Duke-NUS, Graduate Medical School Singapore, 8 College Road, Singapore 169857, Telephone: +65-6516-7666, Email: lorna.fiedler@gmail.com or l.fiedler@imperial.ac.uk.

Blurb

This volume provides a collection of protocols for studying and manipulating VEGF signaling pathways in vitro and in vivo, and in particular, aims to present a range of both firmly established and newly emerging technologies, including those that are amenable to aiding in drug discovery or pre-clinical investigations. Each of the three sections begins with an introductory overview followed by supporting relevant methods. A major advantage of this book is that each chapter not only contains methodological detail rarely seen in other literature, but also contains a section on key notes and troubleshooting advice. Part I focuses on quantification of specific VEGF and VEGF receptor isoforms, beginning with a summary on VEGF splice variants, their function, and their regulation. Part II relates to the study of VEGF dependent signaling in vitro, starting with a comprehensive overview of VEGF-mediated signal transduction in the cardiovascular system that highlights recent discoveries in the field. Part III first details genetic mouse models, then discusses techniques for studying physiological angiogenesis in the developing mouse embryo. 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.

Cutting-edge and thorough, VEFG Signaling: Methods and Protocols is a useful tool for researching and understanding the basic biology of VEGF signaling, and translating it into the clinic.