Receptor Binding Techniques

A comprehensive collection of readily reproducible methods for studying receptors in silico, in vitro, and in vivo. These cutting-edge techniques cover mining from curated databases, identifying novel receptors by high throughput screening, molecular methods to identify mRNA encoding receptors, radioligand binding assays and their analysis, quantitative autoradiography, and imaging receptors by positron emission tomography (PET). Highlights include phenotypic characterization of receptors in knockout mice, imaging receptors using green fluorescent protein and fluorescent resonance energy transfer, and quantitative analysis of receptor mRNA by TaqMan PCR. These book equips the researcher with techniques for exploring the unprecedented number of new receptor systems now emerging and the so-called "orphan" receptors whose activating ligand has not been identified.

This cutting-edge collection of step-by-step experimental protocols demonstrates

One of the major advances in the understanding of the mechanism of action of hormones, neurotransmitters and drugs had arisen from the hypothesis that the physiological or pharmacological responses are trig gered by their interaction with specific cell compo nents, termed receptors. However, the presence of receptors has been inferred from indirect data, and only recently has it been possible to study the kinetics of the interaction between drug and receptors directly, through the so called "binding technique. " This NATO-ASI on "Principles and Methods in Receptor Binding" was devoted mainly to the following aspects of the study of receptors: the principles underlying the use of the binding technique; the mathematical models necessary to interpret binding data; the application of binding methods to specific receptors; and, finally, a few selected examples of coupling between receptors and physiological re sponses. In the chapters of this book, special interest is devoted to the analysis of the simplest models for the interaction of receptors with their ligands (either hormones or neurotransmitters or drugs). The graphical techniques used to analyze the data from binding exper iments are extensively discussed, together with the statistics that have to be used in binding analysis. Moreover, the basic concepts to analyze binding data using a personal computer are presented. v vi PREFACE The factors which should be considered when setting up a binding assay (such as choice of ligand and incubation conditions, preparation of tissue, termina tion of incubation) are discussed.

This book describes the applications of receptor techniques in many different areas in addition to conventional drug and neurotransmitter binding sites. It reviews humoral modulators such as a leukotrienes, interferon, platelet-derived growth factor, and novel endogenous ligands.

First Published in 1982, this book explores the role of receptor-binding radiotracers. Carefully compiled and filled with a vast repertoire of notes, diagrams, and references this book serves as a useful reference for Students of radiology, and other practitioners in their respective fields.

Radiophannaceutical research has recently undergone a major change in direction. In past years it has been concerned mainly with the development of perfusion tracers, the biodistribution of which reflect the regional blood flow to areas of major organs such as the heart and brain. However, a major new direction of interest now lies in the development of receptor-binding radio-tracers which can be used to perform in-vivo characterisation of diseased tissues and it is likely that much of the future research in this field will follow this direction. The difficulties in developing such tracers are considerable. The researcher must first identify a promising target for radiopharmaceutical development. High specific activity radioactive molecules must be designed and synthesised which will both bind to the target receptor with high affinity, and also have the physicochemical characteristics which will allow them to reach the target site in sufficient quantity while at the same time showing minimal uptake in non-target tissues. Thus the knowledge base required for radiophannaceutical development has now expanded beyond the limits of radiopharmaceutical chemistry to include aspects of biochemistry, molecular biology and conventional drug design. The portfolio of basic knowledge required to support current radiopharmaceutical development is changing and scientists working in this arena need to be trained in this regard. At the same time, the very latest developments in the field need to be communicated to the scientific community in order to stimulate the advancement of this exciting new direction of research.

Uniquely integrates the theory and practice of key experimental techniques for bioscience undergraduates. Now includes drug discovery and clinical biochemistry.