A guide for the ECM maze: Methods in Cell-Matrix Adhesion (Methods in Cell Biology, Vol. 69)

edited by Josephine C. Adams
 Academic Press (2002) 461 pages. ISBN 0-12-044142-X $89.95 (paperback)

Studying the extracellular matrix (ECM) and the ways by which cells interact with it is not an easy task. It requires a broad knowledge and practical skills in handling proteins that vary considerably in size, domain organization and post-translational modification. To make matters more complicated, the spatial organization of the ECM proteins holds vital information that is read by cellular adhesion receptors and translated into corresponding cellular behavior. In turn, cells actively modify, reorganize and build up the ECM environment. It is a real experimental challenge to explore, and at the same time to preserve, this tricky balance.

Methods in Cell-Matrix Adhesion is a very helpful guide for scientists daring to accept this challenge and determined to make their way through the ECM maze. The book is part of the very popular Methods in Cell Biology series sponsored by the ASCB and published by Academic Press. In this volume, Josephine Adams has assembled an especially useful collection of protocols written by experts recognized for their contributions in the field of extracellular matrix and cell adhesion.

Part I – Three excellent chapters open the book, outlining briefly the historical development of the field (what we know about the maze), the current appreciation of the three-dimensionality of the ECM and its impact on cellular signaling (where we are now in the maze), and how this research can be translated into practice, e.g. drug discovery (what we can possibly gain by mapping the maze).

Part II – `Matrix Methodologies' comprises eight chapters covering different approaches used to study particular ECM components and particular aspects of the complex extracellular meshwork. They range from classical methods, such as preparation and use of monoclonal antibodies against ECM constituents, to new technologies, such as the use of atomic force microscopy,applied in this case for analysis of matrix dynamics. Separate chapters describe how to purify particular matrix components, how to express them as recombinant proteins by using baculoviruses and how they are proteolytically cleaved, either as individual matrix members or in the context of multicomponent ECM degradation.

Individual ECM proteins become self- or cell-organized into a composite three-dimensional meshwork that functions as an immediate cellular environment. Readers who are interested in these processes can find methods for analysis of basement membrane assembly, combined with extensive background information and protocols for preparation of synthetic or cell-assembled matrices.

Part III – `Adhesion Receptor Methodologies' moves the focus to the cellular receptors: integrins that interact with the ECM. This is the shortest of the three methodological parts, but these chapters are very well written,provide a great deal of specific information and cover most contemporary integrin research. They include strategies for identification of molecules that are involved in cell adhesion, analysis of ligand binding or detection of proteins that associate with extracellular or intracellular domains of integrins. Investigators with a taste for more specialized techniques will find how to use surface plasmon resonance to measure ligand binding and fluorescence resonance energy transfer (FRET) approaches to study the intracellular coupling of adhesion receptors.

Part IV – The last methodological part of the book, `Functional Applications of Cell-Matrix Adhesions in Molecular Cell Biology', covers the approaches used to study cellular responses to different extracellular environments. Classical methods such as cell attachment, cell spreading, and cell migration and invasion assays are very well presented, making them easy to follow and apply. This part also offers some more-specialized approaches like the use of flexible artificial substrates to assess cell-generated traction forces or the use of tissue slices to study cell migration in situ,which mimics more closely the in vivo environment.

In a multiple-author volume like this one, it is important to maintain a good level of coordination between different sections. The editor has kept a good balance between different parts and chapters of the book with one exception. Chapter 18, `Application of Cell Adhesion to Study Signaling Networks', presents protocols that are widely available elsewhere (protein assay, immunoprecipitation, western blotting, etc.). The chapter is presented for novices and differs from the rest of the book, which is developed as a resource for experienced researchers in cell biology. Moreover, some statements, for example, `calcium promotes integrin... cell adhesion' or the suggestion not to boil the SDS-PAGE sample `so as to not reduce the antibody'without mentioning reducing agents such as β-mercaptoethanol, are obviously incorrect.

Apart from this and some other minor overlaps, Methods in Cell-Matrix Adhesion is a very good methodological book that gives answers not only to the `how?' questions but also addresses the equally important `why?'questions, by providing both protocols and background information. It is a useful guide for any laboratory involved in cell-matrix adhesion research, not only as a source of frequently used protocols but also because of its wide methodological scope. The book makes it clear how others approach the complex cell-ECM labyrinth and there is a good chance that it may serve as Ariadne's^* thread, helping daring scientists to navigate the ECM maze successfully.