Gap Junction Protocols

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Direct cell–cell communication is a common property of multicellular organisms that is achieved through membrane channels which are organized in gap junctions. The protein subunits of these intercellular channels, the connexins, form a multigene family that has been investigated in great detail in recent years. It has now become clear that, in different tissues, connexins speak several languages that control specific cellular functions. This progress has been made possible by the availability of new molecular tools and the improvement of basic techniques for the study of membrane channels, as well as by the use of genetic approaches to study protein function in vivo. More important, connexins have gained visibility because mutations in some connexin genes have been found to be linked to human genetic disorders. Connexin Methods and Protocols presents in detail a collection of te- niques currently used to study the cellular and molecular biology of connexins and their physiological properties. The field of gap junctions and connexin research has always been characterized by a multidisciplinary approach c- bining morphology, biochemistry, biophysics, and cellular and molecular biology. This book provides a series of cutting-edge protocols and includes a large spectrum of practical methods that are available to investigate the fu- tion of connexin channels. Connexin Methods and Protocols is divided into three main parts.

The objective in editing this volume was twofold: to provide a reasoned overview of the field as well as to furnish one that provided this overview within the context of the intellectual boundaries of those who initially attempted to define the purview of gap junction research. The latter objective has been realized by selecting the topics for review in this volume. The former objective was achieved by securing the cooperation of leaders in their fields as chapter co-authors.

This monograph contains research articles from the leading laboratories around the world on gap junctions, the intercellular channels that allow communication between neighbouring cells. It describes the latest results in gap junctions research, the structural and functional characterization of cell-cell channels, the assembly and degradation of gap junctions, the expression and function of gap junctions in different tissues, knockout studies of gap junction genes, the regulation of connexin gene expression, and the functions of gap junctions in development and cancer. Any laboratory interested in intercellular communications and gap junction research will find this monograph an indispensable source of information and reference.

Gap junction channels are a group of intercellular channels expressed in tissues and organs to synchronize many physiological processes. A gap junction channel is formed by the docking of two hemichannels, and each hemichannel is a hexamer of connexins. The field of gap junction channel and hemichannel research has recently exploded and became one of the most active areas of cell biology. Numerous novel approaches and techniques have been developed, but there is no single book dedicated to the unique techniques and protocols employed for the research on these large pore channels. This book fills the gap and focuses on protocols, approaches and reviews of gap junction channels and connexin hemichannels. It will be a useful reference for graduate students, postdoctoral fellows and researchers. Anyone with an interest in gap junction channels and hemichannels will need this summary of state-of-the-art techniques and protocols.

Communication between cells via intercellular channels – gap junctions – appears essential to certain developmental processes and appropriate organ function. Gap Junctions in Development and Disease aims to describe the molecular events underlying impaired development and disease. Beginning with a comprehensive review of various mouse and human genes encoding the channel-forming connexins, later chapters describe several connexin mutations associated with human diseases such as hereditary deafness and female infertility. Erroneous signaling mediated by the interaction of mutant connexins with other proteins, thought to be responsible for dysfunction of organs such as heart, muscle, brain, skin, lens, placenta, and endocrine tissue in both mice and men, is also addressed. Although the question of why some mutations in gap-junction proteins lead to specific phenotypes remains to be answered, the reviews in this book provide an intriguing insight into the future direction of this research field.

Gap junctions are present in nearly all tissues, regardless of their embronic origin and have long been of great interest to scientists from many different disciplines. The international meeting on which this book is based brought together 157 scientists from 12 countries and almost as many scientific disciplines. The papers presented at the meeting were reviewed and updated prior to publication in this book. The seven parts of the book progress from general topics to the more specific ones (role of gap junctions in various tissues, regulation and biochemistry, and cancer).