Main description:

12 The average human body has on the order of 10 circulating platelets. They are crucial for hemostasis, and yet excessive platelet activation is a major cause of m- bidity and mortality in Western societies. It is therefore not surprising that platelets have become one of the most extensively investigated biological cell types. We are, however, far from understanding precisely how platelets become activated under physiological and pathophysiological conditions. In addition, there are large gaps in our knowledge of platelet production from their giant precursor cell, the megakar- cyte. Understanding megakaryocyte biology will be crucial for the development of platelet gene targeting. The aim of Platelets and Megakaryocytes is therefore to bring together established and recently developed techniques to provide a comprehensive guide to the study of both the platelet and the megakaryocyte. It consists of five s- tions split between two volumes. The more functional assays appear in Volume 1, whereas Volume 2 includes signaling techniques, postgenomic methods, and a n- ber of key perspectives chapters. Part I of Volume 1, Platelets and Megakaryocytes: Functional Assays, describes many well-established approaches to the study of platelet function, including aggregometry, secretion, arachidonic acid metabolism, procoagulant responses, pla- let adhesion under static or flow conditions, flow cytometry, and production of microparticles. Although one would ideally wish to perform experiments with human platelets, studies within the circulation using intravital microscopy require the use of animal models, which are described in Chapter 16, vol. 1.

Contents:

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