Steroid Receptor Methods

A distinguished team of principal investigators and their associates describe in step-by-step detail a cross-section of the latest research techniques available for studying the endocrine system. As a basis for sophisticated biochemical analysis of receptor properties, the contributors provide methods for the production and purification of a variety of receptors, including progesterone, glucocorticoid, and androgen. Other protocols allow the reader to experiment with DNA binding characteristics, hormone binding assays, and the use of combinatorial chemistry for drug discovery. A series of novel methods utilizing the latest advances in immunochemistry, yeast two-hybrid screening, and fluorescence are included for the detection and analysis of a variety of cellular proteins that influence steroid receptor effectiveness.

Steroid Receptors: Methods and Protocols presents a selection of techniques that have been recently applied to the analysis of steroid receptors, powerful tools for the advancement of our understanding of both the mechanisms regulating gene transcription and the rapid signaling responses of tissues to signals. Research in this area has generated a wealth of data allowing the elucidation of steroid receptor mechanisms and improving the treatment of many endocrine disorders, above all cancers. Chapters cover methods to analyze gene transcription, chromatin and proteomic modifications, extra-nuclear signaling regulation, development of cell and animal models, and preparation of new antibodies. Written in the 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 protocols, and notes on troubleshooting and avoiding known pitfalls. Authoritative and easily accessible, Steroid Receptors: Methods and Protocols offers an updated view on a variety of modern methods that will hopefully contribute to improving our knowledge on the integration of steroid receptors in single or different functionally connected cellular contexts under a variety of physiological and pathological conditions, above all tumors.

These volumes represent the "state-of-the-art" in morphological methods for detection of steroid receptors. Some physician authors have applied many of the new methodologies in clinical research projects deisgned to improve medical practice, and these attempts have been placed in Volume II. The Background and recent advances presented here are intended to stimulate further experimentation and new innovations, and spur clinical trials by all investigators with an interest in steroid hormones, cell biology, and clinical oncology.

During the last two decades, progress in steroid hormone research has resulted in the development of new approaches to contraception as well as diagnosis and treatment of endocrine disorders and cancers. Although significant advances have been made in the purification, characterization, immunochemistry and molecular biology of steroid receptors, the precise molecular mechanism of steroid hormone action has remained obscure. This book captures the detailed presentations made at the first conference on Steroid Receptors in Health and Disease held at Meadow Brook Hall, Oakland University in the fall of 1987. The purpose of this international con ference was to facilitate scientific exchange toward a better understand ing of the mode of action of steroid hormones. The scientific sessions consisted of poster presentations and state-of-the-art lectures, the latter of which make up this volume. The first chapter is meant to provide the reader with a more general background of the topics covered in the book, as well as to discuss certain theme-related issues that are either not yet well-established or accepted or are in the stage of infancy. It is hoped that this volume will serve as a useful treatise for students and investigators interested in basic and clinical aspects of biological regulation by steroid hormones. A task of this magnitude could not have been undertaken without the encouragement, advice and continued generous assistance of the members of the scientific committee. I am gratefully indebted to Drs.

The past few years have witnessed the emergence of steroid hormones as the wonder molecules which generate as much discussion in the scientific literature as they do in a typical living room. This transition has been a result of the tremendous public and scientific interest in the normal functioning of the hor mones as well their suggested involvement in several clinical conditions. In the recent past, notable scientific and technological advances have been made in the areas of contraception and regulation of fertility. Steroid receptors are the indis pensable mediators of hormonal responses and are complex protein molecules which appear to exist in association with other, yet undefined, proteins and/or factors. Receptors for vitamin D, retinoic acid and the thyroid hormones share structural similarities with steroid receptors, and the roster of this superfamily is still expanding. While our knowledge of the diversity and magnitude of steroid effects has advanced, the precise mode of steroid hormone action has alluded investigators. This volume brings together an international team of prominent investigators who discuss their most recent work on the basic and clinical aspects of steroid/nuclear receptors. The contributions represent updated versions of the invited presentations made at The Second Meadow Brook Conference on Steroid Receptors in Health and Disease. I am grateful to my colleagues on the Scientific Committee: Etienne Baulieu, Jack Gorski, Benita Katzenellenbogen, David Toft and James WittJiff, who provided the vision and guidance in formulating an out standing program.

Steroid hormones are unique compounds in that they are active at the interface of peripheral endocrine events and neural mechanisms. Thus their effects present an important peripheral signaling system to alter brain function. This volume presents state-of-the-art and classical techniques for the study of steroid hormones and their receptors and their effects and actions. Comprehensive protocols included for the study of Steroid kinetics and metabolism Steroid receptors Molecular and cellular effects of steroids Steroid effects on integrated systems

The papers in this volume were presented at the Symposium on Steroid Hormone Receptor Systems held October 18-20, 1978, at the Worcester Foundation for Experimental Biology, Shrewsbury, Mass. The meeting was organized to review, discuss, and disseminate new knowledge about the regulation and function of the receptor proteins which mediate estrogen, progestin, glucocorticoid, and androgen action. The symposium brought together leading scientists whose interests span the spectrum of biological organization. On this occasion, Drs. Elwood V. Jensen and Etienne E. Baulieu were honored as recipients of the Tenth Annual Gregory Pincus l1emorial Award for their pioneering studies of steroid hormone receptors. The material covered in this book focuses on the molecular mechanisms which control receptor site availability and function. The topics discussed include recent work on receptor antibodies, antiestrogen action, receptor heterogeneity, nuclear binding and processing of receptor, receptor activation and inactivation mechan isms, interactions between receptor systems, influence of biological rhythms, membrane binding sites, and the evolution of steroid-bind ing proteins. Although studies on steroid receptors have led to the development of new approaches for cancer therapy and contraception, much remains to be discovered in this rapidly expanding field. Hope fully, this book will provide added impetus to the quest for a full understanding of steroid receptor systems by drawing attention to the unresolved questions yet to be answered. Hhile the book is intended primarily for those interested in hormone action, it should be of value to a more general audience including cell, molecular, and developmental biologists.