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
José Rivera, María 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 José A. López

Measurement and Manipulation of [Ca2+]i in Suspensions of Platelets and Cell Cultures
Philippe Ohlmann, Béatrice 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

Platelets are an essential element of the body's hemostasis system, and yet, through their involvement in thrombosis, are also a major cause of morbidity and mortality. Platelets have a limited lifespan and must be continually manufactured by their precursor, the megakaryocyte. In Platelets and Megakaryocytes, Volume 2: Perspectives and Techniques leading expert researchers-many responsible for seminal discoveries in the field-offer basic and advanced techniques for studying cell signaling in the regulation and function of 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 in the regulation of cell signaling, and the measurement of intracellular calcium levels. There are also techniques for the study of electrophysiological events, nitric oxide signaling, venom proteins, and for the internalization of proteins into platelets through permeabilization. The application of many molecular and post-genomic methods to the study of platelets and megakaryocytes is also described. All protocols follow the successful Methods in Molecular Biology(TM) 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. A companion volume, Platelets and Megakaryocytes, Volume 1: Functional Assays describes in step-by-step detail basic and advanced techniques for analyzing platelet and megakaryocyte function.
Comprehensive and cutting-edge, the two volumes of Platelets and Megakaryocytes 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.