In this cutting-edge book, internationally renowned experts present techniques which reflect many of the recent technological advances in experimental tools for cytoskeleton research. There is emphasis on animal, plant, protist, and fungal model systems.

The third edition of this volume focuses on experimental models that are useful for investigating various aspects of cytoskeleton structure and function. Animal, plant, protist, and fungal models highlight twenty-four chapters that provide detailed protocols for live and fixed-cell imaging, dynamics of cytoskeleton components, cell and organelle motility, and genetics and proteomics. Written in the highly successful Methods in Molecular Biology series format, protocols in each chapter are up-to-date menus organized in a useful step-by-step format appropriate for novice and established investigators. Each chapter is equipped with a valuable notes section that provides a troubleshooting guide and helpful, and often unpublished, technical information aimed at ensuring success with implementation of the protocols. Authoritative and thorough, Cytoskeleton Methods and Protocols, Third Edition helps researchers expand their understanding of cytoskeleton structure and function.

This detailed volume explores the development of technologies and protocols that are currently being used to understand the nature and activities of the plant cytoskeleton. A focus for many of the chapters is on sample preparation, as the quality of plant organ/tissue preparation, from single to multicellular samples, determines the quality of the data. Written for the highly successful Methods in Molecular Biology series, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step and readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls. Authoritative and practical, The Plant Cytoskeleton: Methods and Protocols serves as an ideal guide for researchers interested in or starting to be interested in plant cell and molecular biology research.

This volume details comprehensive state-of-the-art methods on actin microfilaments and microtubules and how they work to achieve different cellular functions in different cellular contexts. Chapters guide readers through protein purification, in vitro reconstitution of several cytoskeleton properties, analyses of microtubule- and actin-based structures, functional dissection of post-translational modifications, and roles in several biological processes. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls. Authoritative and cutting-edge, Cytoskeleton Dynamics: Methods and Protocols aims to provide a wide range of experimental approaches and be an invaluable resource for present and future generations of cytoskeleton researchers. The chapter “Visualization and Functional Analysis of Spindle Actin and Chromosome Segregation in Mammalian Oocytes” is available open access under a Creative Commons Attribution 4.0 International License via link.springer.com.

Over the past two decades experimental studies have solidified the int- pretation of the cytoskeleton as a highly dynamic network of microtubules, actin microfilaments, intermediate filaments, and myosin filaments. Rather than a network of disparate fibers, these polymers are often interconnected and display synergy, which is the combined action of two or more cytoskeletal polymers to achieve a specific cellular structure or function. Cross-commu- cation among cytoskeletal polymers is thought to be achieved through cytoskeletal polymer accessory proteins and molecular motors that bind two or more cytoskeletal polymers. Development of the modern concept of the cytoskeleton is a direct o- growth of advances in experimental tools and reagents that are available to cell and molecular biologists. Technological advances and refinements in cell imaging have made it possible to selectively image a single cytoskeletal po- mer and monitor its dynamics through the use of fluorescence probes in vitro and in vivo. Two decades ago, cytoskeletal research was limited to a few perturbation reagents that included colchicine and cytochalasin. Today, the perturbation arsenal has expanded to a highly selective group of reagents that includes Taxol, nocodazole, benomyl, latrunculin, jasplakinolide, and such endogenous proteins as gelsolin. These reagents enable the investigator to selectively perturb or destroy a cytoskeletal polymer while leaving other cytoskeletal polymers intact. Site-specific monoclonal antibodies that target a specific cytoskeletal polymer have proven to be highly selective affinity tools for cytoskeletal research.

This book provides descriptions of experimental methods in research on the cytoskeleton and its relationships to signaling and cell regulation. Thus, it bridges two active and fertile areas of research. The focus is directed particularly towards methods which take advantage of recent advances in molecular biology, microscopy and immunological assays. A second emphasis is on methods for understanding dynamic changes in cells. A third emphasis is on the formation and turnover of macromolecular and supramolecular complexes, which are so important in driving cell regulation and the behaviour of cytoskeletal elements. A combination of practical advice and detailed protocols should make this book valuable for both novice and experienced workers in these burgeoning fields.