Platelets and Megakaryocytes: Volume 2: Perspectives and Techniques (Methods in Molecular Biology)

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.

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™ 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.