Exocytosis And Endocytosis

In this book, skilled experts provide the most up-to-date, step-by-step laboratory protocols for examining molecular machinery and biological functions of exocytosis and endocytosis in vitro and in vivo. The book is insightful to both newcomers and seasoned professionals. It offers a unique and highly practical guide to versatile laboratory tools developed to study various aspects of intracellular vesicle trafficking in simple model systems and living organisms.

This volume provides basic and cutting-edge methods and protocols to study the major characteristics of eukaryotic cells. Chapters detail the different pathways of endocytosis in vivo, real time imaging of endocytic steps, endocytosis in model organisms, super-resolution methods to follow proteins involved in exocytosis, specific protocols for exocytosis in specialized cells such as neutrophils or neuroendocrine cells, as well as secretion of exosomes. 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. Cutting-edge and clearly written, Exocytosis and Endocytosis: Methods and Protocols is a valuable resource for researchers in the fields of cell biology, neurology, immunology, oncology, and those interested in studying protein trafficking and signal regulation.

Volume 4 of Biomembranes covers endocytosis, exocytosis and related processes. A major role of the plasma membrane is as a permeability barrier, keeping the inside of the cell inside and the outside, outside. Mechanisms must then exist to allow movement of material between the cell and its environment. One mechanism for export from the cell is by exocytosis, a process in which the membranes of secretory vesicles fuse with the plasma membrane releasing the contents of the vesicle into the extracellular medium. The process has been studied in particular depth for the release of neurotransmitters at the synapse. Import into the cell is possible by the process of receptor-mediated endocytosis in which selected plasma membrane proteins are internalizes; when these proteins are receptors for macromolecules, the result is uptake of the macromolecule. Transferring, the low-density lipoprotein, and asialoglycoproteins are all taken up into cells in this way. Phagocytosis, the ingestion of cells and cell fragments by neutrophils and macrophages, also involves receptors - on the phagocytic membrane - of which the best studied are those for the Fc domain of IgG, for the third component of complement, and for the mannose/fructose carbohydrates. Protection of a host against infection can also be achieved by damaging the integrity of the plasma membrane of the invading organism. This is the strategy evolved by the cytotoxic T lymphocytes, which produce a pore-forming toxin, perforin. Volume 4 of Biomembranes explores the structures and mechanisms involved in these biologically and medically important processes.

The transport of macromolecules in membrane bounded vesicles is a fundamental process of all eukaryotic cells. The paths taken by this vesicular traffic have been intensively researched in animal cells but are less well characterised in plant cells. Nevertheless, with the development of and combination of techniques in biochemistry, molecular biology and cell biology, progress in the study of plant vesicle traffic has been achieved in recent years. This book attempts to highlight the recent advances made and to explore avenues for future research. The book opens with a general overview of vesicular traffic both in animal and plant cells. This is followed by a more detailed consideration of higher plant coated vesicles, endocytosis, exocytosis and mechanisms of vesicle traffic and transport in cells. The biochemistry, cell biology and molecular biology of each is considered and particular attention is given to those specialised cell systems in which vesicle traffic is of particular importance. This book brings together expertise from a wide range of research disciplines involved in the study of vesicle traffic in plants, and, as such, provides a unique synthesis from which the research can move forward. It will be of general interest to those involved in the study of vesicle traffic in both plant and animal cells.

This volume of the treatise deals with structural aspects of the cytoskeleton: the characteristics of the filaments and their components; the organization of the genes; motor proteins; interactions with membranes.

Endocytosis is a fundamental cellular process by means of which cells internalize extracellular and plasma membrane cargos for recycling or degradation. It is important for the establishment and maintenance of cell polarity, subcellular signaling and uptake of nutrients into specialized cells, but also for plant cell interactions with pathogenic and symbiotic microbes. Endocytosis starts by vesicle formation at the plasma membrane and progresses through early and late endosomal compartments. In these endosomes cargo is sorted and it is either recycled back to the plasma membrane, or degraded in the lytic vacuole. This book presents an overview of our current knowledge of endocytosis in plants with a main focus on the key molecules undergoing and regulating endocytosis. It also provides up to date methodological approaches as well as principles of protein, structural lipid, sugar and microbe internalization in plant cells. The individual chapters describe clathrin-mediated and fluid-phase endocytosis, as well as flotillin-mediated endocytosis and internalization of microbes. The book was written for a broad spectrum of readers including students, teachers and researchers.