VEGF Signaling: Methods and Protocols

ISBN-13: 9781071622193 / Angielski

Lorna R. Fiedler; Caroline Pellet-Many

Opis

This volume provides an updated collection of protocols for manipulating and studying VEGF signaling pathways in vitro and in vivo and aims to present a range of both firmly established and newly emerging technologies. Covering multiple model species, from mouse to zebrafish to human, the book explores the role of VEGF and VEGFR isoforms in exosomes, cultured cells, or in tissues, as well as robust cell assays for the investigation of basic angiogenic mechanisms and VEGF signaling in more complex cellular systems, amongst other subjects. 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. 

Authoritative and up-to-date, VEGF Signaling: Methods and Protocols, Second Edition provides a useful tool for researchers in the vascular biology community and beyond in understanding the basic biology of VEGF signaling and in translating this research into the clinic.

Spis treści

1. Opposing Roles of Vascular Endothelial Growth Factor C in Metastatic Dissemination and Resistance to Radio/Chemotherapy: Discussion of Mechanisms and Therapeutic Strategies

            Christopher Montemagno, Frédéric Luciano, and Gilles Pagès

 

2. Identification of VEGF Isoforms in Mouse, Rat, and Zebrafish Using RT-qPCR

            Patricia de Winter

 

3. Multi-Parameter Fluorescence-Activated Cell Sorting of Human Lymphatic Endothelial Cells

            Theresa Connor, Nerida Sleebs, and Zerina Lokmic-Tomkins

 

4. Absolute Quantification of Plasma Membrane Receptors via Quantitative Flow Cytometry

            Yingye Fang, Manasi Malik, Sarah K. England, and P.I. Imoukhuede

 

5. Measurement of VEGF Content in Exosomes and Subsequent Tumor Tubulogenesis and In Vivo Angiogenesis Functional Assays

            Ye Zeng, Xiaoqiang Du, Wenli Jiang, and Yan Qiu

 

6. SH2-Domain Protein Isolation Using Synthetic Phosphorylated Peptides to Study VEGFR2 Signaling

            Chiara Testini

 

7. Monitoring VEGF-Stimulated Calcium Ion Flux in Endothelial Cells

            William R. Critchley, Gareth W.F. Fearnley, Izma Abdul-Zani, Carmen Molina-Paris, Claus Bendtsen, Ian C. Zachary, Michael A. Harrison, and Sreenivasan Ponnambalam

 

8. Co-Immunoprecipitation Assays

            Ian M. Evans and Ketevan Paliashvili

 

9. Using Immortalized Endothelial Cells to Study the Roles of Adhesion Molecules in VEGF-Induced Signaling

            James A.G.E. Taylor, Christopher J. Benwell, and Stephen D. Robinson

 

10. RNAscope for VEGF-A Detection in Human Tumor Bioptic Specimens

            Tiziana Annese, Roberto Tamma, and Domenico Ribatti

 

11. Global MicroRNA Profiling of Vascular Endothelial Cells

            Eloi Schmauch, Anna-Liisa Levonen, and Suvi Linna-Kuosmanen

 

12. Endothelial Cell Tube Formation Assay: An In Vitro Model for Angiogenesis

            Mary Kelley, Sara Fierstein, Laura Purkey, and Kathleen DeCicco-Skinner

 

13. Use of a Thin Layer Assay for Assessing the Angiogenic Potential of Endothelial Cells In Vitro

            James A.E. Lane, Ashton Faulkner, Elizabeth J.T. Finding, Eleanor G. Lynam, and Caroline P.D. Wheeler-Jones

 

14. VEGF-A₁₆₅ -Induced Endothelial Cells Chemotactic Migration and Invasion Assays

            Caroline Pellet-Many

 

15. Measuring Mitochondrial Calcium Fluxes in Cardiomyocyte upon Mechanical Stretch-Induced Hypertrophy

            Daniela Ramaccini, Carlotta Giorgi, and Michelle L. Matter

 

16. Simultaneous Measurement of Endothelial Cell Proliferation and Cell Cycle Stage Using Flow Cytometry

            Eleanor G. Lynam, James A.E. Lane, Elizabeth J.T. Finding, and Caroline P.D. Wheeler-Jones

 

17. Ex Vivo Mouse Aortic Ring Angiogenesis Assay

            Vedanta Mehta and Marwa Mahmoud

 

18. Retinal Microvasculature-on-a-Chip for Modeling VEGF-Induced Permeability

            Héloïse Ragelle, Karen Dernick, Peter D. Westenskow, and Stefan Kustermann

 

19. Preventing VEGF-Mediated Vascular Permeability by Experimentally Potentiating BBB Characteristics in Endothelial Cells

            Bo Kyoung Kim, Jérémie Canonica, Filip Roudnicky, and Peter D. Westenskow

 

20. The Embryonic Mouse Hindbrain and Postnatal Retina as In Vivo Models to Study Angiogenesis

            Alessandro Fantin and Christiana Ruhrberg

 

21. Evaluating VEGF-Induced Vascular Leakage Using the Miles Assay

            James T. Brash, Christiana Ruhrberg, and Alessandro Fantin

 

22. Modulation of VEGFA Signaling during Heart Regeneration in Zebrafish

            Kaushik Chowdhury, Shih-Lei Lai, and Rubén Marín-Juez

 

23. Three-Dimensional Visualization of Blood and Lymphatic Vessels in the Adult Zebrafish Heart by Chemical Clearing

            Lorna R. Fiedler, Paul R. Riley, and Roger Patient

 

24. Fluorescence-Activated Cell Sorting and Quantitative Real-Time PCR to Reveal VEGF Expressing Macrophage Populations in the Zebrafish Larvae

            Andrew Herman, Alexander Greenhough, and David B. Gurevich

 

25. Assessing Molecular Regulation of Vascular Permeability Using a VEGF-Inducible Zebrafish Model

            Luke H. Hoeppner