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MODULATING GENE EXPRESSION BY ANTISENSE OLIGONUCLEOTIDES TO UNDERSTAND NEURAL FUNCTIONING THE KLUWER SERIES PERSPECTIVES IN ANTISENSE SCIENCE With this volume, Kluwer Academic Publishers inaugurates the first in a series of in-depth volumes that discuss specific applications of the antisense biotechnology. Several books that briefly cover these subjects, which include uses of antisense oligomers in restenosis and leukemia are currently available; however, none devote such extensive coverage to what are highly complex, often controversial issues in each area of study. As Dr. McCarthy appropriately indicates in her chapter, antisense is a complex business. The proper interpretation of experimental results, may be particularly difficult, expecially when the phosphorothioate backbone is used, as it frequently is. These problems are due (in part) to non-sequence specific interactions of the phosphorothioates with cellular and extra cellular proteins, particularly those with heparin-binding sites. The difficulties are exacerbated when oligomers longer than 18-20-mer are used (in the absence of a delivery vehicle) at concentrations greater than about 5 flM. In such cases, observed experimental results are almost certainly due to a combination of sequence-specific and non-sequence specific effects. This is particularly true when phosphorothioates bearing a 3' or 5' GGGG sequence motif are used. These molecules can form tetraplexes, which have interesting, but a priori unpredictable effects on cells. Thus, claims of sequence specific antisense activity must always be evaluated in the context of these caveats, and in my opinion a health skepticism should usually be maintained. Nevertheless, this volume represents an invaluable comependium of the neurosciences antisense literature, and provides ample material for thoughtful reflection. C. A. Stein, Series Editor MODULATING GENE EXPRESSION BY ANTISENSE OLIGONUCLEOTIDES TO UNDERSTAND NEURAL FUNCTIONING edited by Margaret M. McCarthy University ofM aryland, Baltimore PERSPECTIVES IN ANTISENSE SCIENCE Series Editor: C. A. Stein The College ofP hysicians & Surgeons, Columbia University 1IIrI... " SPRINGER SCIENCE+BUSINESS MEDIA, LLC Library of Congress Cataloging-in-Publication Data Modulating gene expression by antisense oligonucleotides to understand neural functioning / edited by Margaret M. McCarthy. p. cm. Includes bibliographical references and index. ISBN 978-1-4613-7238-7 ISBN 978-1-4615-4933-8 (eBook) DOI 10.1007/978-1-4615-4933-8 1. Molecular neurobiology. 2. Antisense nucleic acids. 3. Genetic regulation. 1. McCarthy, Margaret M., 1958- QP356.2.M58 1998 572.8'65--dc21 98-44227 CIP Copyright © 1998 Springer Science+Business Media New York Originally published by Kluwer Academic Publishers in 1998 Softcover reprint ofthe hardcover Ist edition 1998 AlI rights reserved. No part of this publication may be reproduced, stored in a retrieval system or transmitted in any form or by any means, mechanical, photo copying, recording, or otherwise, without the prior written permission of the publisher, Springer Science+Business Media, LLC . Printed an acid-free paper. CONTENTS Contributors ......................................v ii Acknowledgements .................................x i 1 . The Use of Antisense Technologies to Understand Neural Functioning: Fact or Fiction? Margaret M. McCarthy ............................ 1 2. Tissue Distribution, Cellular Uptake, and Intracellular Localization and Stability of Centrally Administered Oligonucleotides in the Brain: Implications for Behavioral and Physiological Effects of Antisense Oligonucleotides Wolfgang Sommer, Donald W. Pfaff, and Sonoko Ogawa .... 9 3 . Non-Specific Effects of Centrally Administered Oligonucleotides Rainer Spanagel, Bernd Schobitz, and Mario Engelmann .....2 7 4 . Rapid Onset of Antisense Effects: Evidence for a Close Link Between Gene Expression and Neuronal Activity Inga D. Neumann and Quentin J. Pittman ...............4 3 5 . Effects of Dopamine Receptor Antisense RNA Expression Vectors in the Nervous System Genoveva Davidkova, Sui-Po Zhang, Long-Wu Zhou, and Benjamin Weiss ................................6 1 6. Modification of Native GABA Receptor Assemblies A Using Antisense Oligonucleotides Dennis R. Grayson and Stephano Vicini ................8 3 7 . Antisense Oligodeoxynucleotides as Specific Tools for Studying Opioid Receptor-Mediated Analgesia Leon F. Tseng ................................1 03 VI 8 . Antisense Approaches to Investigate the Neuroendocrine Control of Thirst and Sodium Appetite in the Rat Randall R. Sakai, Li Yun Ma, and Steven J. Fluharty ......1 25 9 . Steroid-Mediated Behaviors and their Regulation by Antisense Oligodeoxynucleotides Shailaja K. Mani and Pramod K. Dash ................1 41 10. Modulation of Endocrine and Behavioral Rhythms with Antisense Oligonucleotides Kathryn Scarbrough ............................ 159 Index .......................................... 171 CONTRIBUTORS Pramod K. Dash Department of Neurobiology and Anatomy University of Texas-Houston Health Science Center Houston, TX 77225, USA Genoveva Davidkova Department of Pharmacology MCP Hahnemann School of Medicine Allegheny University of the Health Sciences Philadelphia, PA 19129, USA Mario Engelmann Department of Neuroendocrinology Max Planck Institute of Psychiatry KRAEPELINSTRASSE 2 80804 Munich, GERMANY Steven J. Fluharty Department of Animal Biology School of Veterinary Medicine University of Pennsylvania Philadelphia, PA 19104, USA Dennis R. Grayson Center for Neurosciences Research Allegheny University of Health Sciences Pittsburgh, PA 15212, USA current address: Department of Psychiatry (MC 912) The Psychiatric Institute 1601 West Taylor Street Chicago, Illinois 60612 Li Yun Ma Department of Animal Biology School of Veterinary Medicine University of Pennsylvania Philadelphia, PA 19104, USA viii Shailaja K. Mani Department of Cell Biology Baylor College of Medicine Houston, TX 77030, USA Margaret M. McCarthy Department of Physiology School of Medicine University of Maryland, Baltimore Baltimore, MD 21201, USA Inga D. Neumann Department of Neuroendocrinology Max Planck Institute of Psychiatry KRAEPELINSTRASSE 2 80804 Munich, GERMANY Sonoko Ogawa Laboratory of Neurobiology and Behavior The Rockefeller University New York, NY 10021, USA Donald W. Pfaff Laboratory of Neurobiology and Behavior The Rockefeller University New York, NY 10021, USA Quentin J. Pittman Neuroscience Research Group and Department of Medical Physiology University of Calgary Calgary, AB T2N 4Nl, CANADA Randall R. Sakai Department of Animal Biology School of Veterinary Medicine University of Pennsylvania Philadelphia, PA 19104, USA Kathryn Scarbrough Department of Neurobiology and Physiology Northwestern University Evanston, IL 60208, USA ix Bernd Schiibitz Department of Neuroendocrinology Max Planck Institute of Psychiatry KRAEPELINSTRASSE 2 80804 Munich, GERMANY Wolfgang Sommer Department of Clinical Neuroscience Karolinska Institute Stockholm S-17177, SWEDEN Rainer Spanagel Department of Neuroendocrinology Max Planck Institute of Psychiatry KRAEPELINSTRASSE 2 80804 Munich, GERMANY Leon F. Tseng Department of Anesthesiology Medical College of Wisconsin Milwaukee, WI 53226, USA Stephano Vicini Departments of Physiology and Biophysics Georgetown University School of Medicine Washington, DC 20007, USA Benjamin Weiss Department of Pharmacology MCP Hahnemann School of Medicine Allegheny University of the Health Sciences Philadelphia, PA 19129, USA Sui-Po Zhang Department of Pharmacology MCP Hahnemann School of Medicine Allegheny University of the Health Sciences Philadelphia, PA 19129, USA Long-Wu Zhou Department of Pharmacology MCP Hahnemann School of Medicine Allegheny University of the Health Sciences Philadelphia, PA 19129, USA ACKNOWLEDGEMENTS The editor would like to thank Ms. Rachel Dvoskin for her outstanding expert editorial and technical assistance and without whom this volume would not have been possible. She also thanks Dr. Gregory F. Ball for his unfailing support and encouragement and NIH grant MH52716 for financial support.

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by Margaret M. McCarthy (auth.), Margaret M. McCarthy (eds.)

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