For both volumes: A comprehensive collection of cutting-edge methods for the molecular and cellular analysis of platelets and megakaryocytes. Volume 1 details basic and advanced techniques for analyzing platelet and megakaryocyte function. The approaches presented for platelet analysis include aggregometry, secretion, arachidonic acid metabolism, procoagulent response, platelet adhesion under static and flow conditions, flow cytometry, and production of microparticles. The techniques for study of megakaryocyte function allow the measurement of specialized structures for platelet production, the appearance of platelet-specific surface receptors, and the increase in ploidy. Volume 2 offers techniques for studying many aspects of signaling in platelets and megakaryocytes, as well as state-of-the-art commentaries on our understanding of these cells. The methods include ligand-binding assays, the study of protein and lipid kinases and phosphatases, the analysis of lipid rafts, and the measurement of intracellular calcium levels.
There are also techniques for study of electrophysiological events, nitric oxide signaling, venom proteins, and for the internalization of proteins into platelets through permeabilization. This volume also describes the application of molecular and post-genomic methods are to the study of platelets and megakaryocytes. All protocols follow the successful Methods in Molecular Biology(t) series format, each one offering step-by-step laboratory instructions, an introduction outlining the principle behind the technique, lists of equipment and reagents, and tips on troubleshooting and avoiding known pitfalls. The two volumes offer novice and experienced cell biologists, hematologists, and clinicians not only a benchmark survey of the field, but also a comprehensive library of proven techniques essential for productive research on platelets and megakaryocytes today.
Part I. Perspectives Signaling Receptors on Platelets and Megakaryocytes Donna Woulfe, Jing Yang, Nicolas Prevost, Peter O'Brien, Ryan Fortna, Massimiliano Tognolini, Hong Jiang, Jie Wu, and Lawrence F. Brass Manipulation of Gene Expression in Megakaryocytes Nikla R. Emambokus, George J. Murphy, and Jonathan Frampton In Vitro Changes of Platelet Parameters: Lessons From Blood Banking Jose Rivera, Maria L. Lozano, and Vicente Vicente Pharmacological Approaches to Studying Platelet Function: An Overview Susanna M. O. Hourani Inhibitors of Cellular Signaling Targets: Designs and Limitations Chris J. Vlahos and Louis F. Stancato Part II Signaling Techniques. Ligand-Binding Assays for Collagen Stephanie M. Jung and Masaaki Moroi Use of Radiolabeled 2-Methylthio-ADP to Study P2Y Receptors on Platelets and Cell Lines Pierre Savi and Jean-Marc Herbert Ligand-Binding Assays: Fibrinogen Dermot Cox Techniques for the Analysis of Proteins by SDS-Polyacrylamide Gel Electrophoresis and Western Blotting Jonathan M. Gibbins Study of Tyrosine Kinases and Protein Tyrosine Phosphorylation Jonathan M. Gibbins Protein Tyrosine Phosphatases Matthew L. Jones and Alastair W. Poole The Study of Serine-Threonine Kinases Gertie Gorter and Jan Willem Akkerman Phosphoinositides: Lipid Kinases and Phosphatases Bernard Payrastre Isolation and Analysis of Platelet Lipid Rafts Corie N. Shrimpton, Karine Gousset, Fern Tablin, and Jose A. Lopez Measurement and Manipulation of [Ca2+]i in Suspensions of Platelets and Cell Cultures Philippe Ohlmann, Beatrice Hechler, Jean-Pierre Cazenave, and Christian Gachet Measurement and Manipulation of Intracellular Ca2+ in Single Platelets and Megakaryocytes Michael J. Mason and Martyn P. Mahaut-Smith Patch-Clamp Recordings of Electrophysiological Events in the Platelet and Megakaryocyte Martyn P. Mahaut-Smith Evaluation of Nitrotyrosine-Containing Proteins in Blood Platelets Khalid M. Naseem, Rocio Riba, and Max Troxler Nitric Oxide Signaling in Platelets Sylvia Y. Low and K. Richard Bruckdorfer Snake Venom Toxins Affecting Platelet Function Robert K. Andrews, Elizabeth E. Gardiner, and Michael C. Berndt Peptide Synthesis in the Study of Collagen-Platelet Interactions C. Graham Knight, Catherine M. Onley, and Richard W. Farndale Platelet Permeabilization Robert Flaumenhaft Part III. Molecular and Post-Genomic Techniques Using Retroviruses to Express Genes in Primary Megakaryocyte Lineage Cells Meenakshi Gaur, George J. Murphy, Jonathan Frampton, and Andrew D. Leavitt Use of Antisense Oligonucleotide Technology to Investigate Signaling Pathways in Megakaryocytes Hava Avraham, Shalom Avraham, and Radoslaw Zagozdzon GFP Fusion Proteins to Study Signaling in Live Cells Simon A. Walker, Gyles E. Cozier, and Peter J. Cullen Two-Dimensional Polyacrylamide Gel Electrophoresis for Platelet Proteomics Katrin Marcus and Helmut E. Meyer Preparation of mRNA and cDNA Libraries From Platelets and Megakaryocytes Benjamin Z. S. Paul, Jiango Jin, and Satya P. Kunapuli Platelet Receptor Structures and Polymorphisms Thomas J. Kunicki, Steven Head, and Daniel R. Salomon Gene Array Technology and the Study of Platelets and Megakaryocytes Lloyd T. Lam and Emery H. Bresnick Index