Hydra: Research Methods Hydra: Research Methods Edited by Howard M. Lenhoff University of California Irvine, California SPRINGER SCIENCE+BUSINESS MEDIA, LLC Library of Congress Cataloging in Publication Data Main entry under title: Hydra: research methods. lncludes bibliographical references and index. 1. Hydra. 1. Lenhoff, Howard M. QL377.H9H93 1982 593.7'1 82-24648 ISBN 978-1-4757-0598-0 ISBN 978-1-4757-0596-6 (eBook) DOI 10.1007/978-1-4757-0596-6 Cm·cr photo courtesy of Regula Bănninger and Prof. Pierre Tardent, Zoologica! Institute, University of Zurich, Switzerland. <c; 1983 Springer Scicncc+Busincss Media New York Originally publishcd by Plcnum Prcss, New York in 1983 Ali rights reserved No part of this book may be reproduced, stored in a retrieval system, or transmitted in any form or by any means, electronic, mecilanical, photocopying, microfilming, recording, or otherwise, without written permission from the Publisher Contributors Richard S. Blanquet, Department of Biology, Georgetown University, Washington, D.C. 20057 Hans R. Bode, Developmental Biology Center and Department of Developmental and Cell Biology, University of California, Irvine, California 9271 7 Patricia M. Bode. Developmental Biology Center and Department of Developmental and Cell Biology, University of California, Irvine, California 9271 7 Richard D. Campbell, Department of Developmental and Cell Biology, University of California, Irvine, California 92717 Jean Danner, Biochemistry Section, NIOSH, Morgantown, West Virginia 26505 Charles N. David, Department of Molecular Biology, Albert Einstein College of Medicine, Bronx, New York 10461 Robert M. Day, Department of Developmental and Cell Biology, University of California, Irvine, California 92717 John F. Dunne, Developmental Biology Center, University of California, Irvine, California 92717 Kristine M. Flick. Developmental Biology Center and Department of Developmental and Cell Biology, University of California, Irvine, California 92717 v vi Contributors Cheng-Mei Fradkin, Department of Developmental and Cell Biology, University of California. Irvine, California 92717 Cornelius J. P. Grimmelikhuijzen, Max-Pianck-Institut fUr medizinische Forschung, Abteilung Biophysik, 6900 Heidelberg, Federal Republic of Germany ~vrta Heagy, Department of Microbiology, University of Massachusetts. Amherst, Massachusetts 01003 David A. Hessinger, Department of Physiology and Pharmacology, School of Medicine, Lorna Linda University, Lorna Linda, California 92350 Jeanne Ivy, Department of Developmental and Cell Biology, University of California, Irvine California 92717 Robert K. Josephson, School of Biological Science, University of California, Irvine, California 92717 EdwardS. Kline, Department of Biochemistry, Medical College of Virginia, Virginia Commonwealth University, Richmond, Virginia 23298 Howard M. Lenhoff, Department of Developmental and Cell Biology. University of California, Irvine, California 92717 Georgia E. Lesh-Laurie, Department of Biology, Cleveland State University, Cleveland, Ohio 44115 C. Lynne Littlefield, Developmental Biology Center, University of California, Irvine, California 92717 Stephen S. Macintyre, Department of Anatomy, Case Western Reserve University, Cleveland, Ohio 44106 Harry K. MacWilliams, Department of Anatomy, University of Massachusetts Medical Center, Worcester, Massachusetts 01605 Bever(v A. Marcum, Department of Biological Sciences, California State University, Chico, California, 95929 L. Muscatine, Department of Biology, University of California, Los Angeles, California 90024 Contributors vii That T. Ngo, Department of Developmental and Cell Biology, University of California, Irvine, California 92717 Patricia Novak, Department of Developmental and Cell Biology, University of California, Irvine, California 92717 Joann J. Otto, Department of Biological Sciences, Purdue University, West Lafayette, Indiana 47907 R. L. Pardv, School of Life Sciences, University of Nebraska, Lincoln, Nebraska 68588 Donald W Phipps, Jr., School of Life Sciences, University of Nebraska, Lincoln, Nebraska 68588 M. Rahat, Department of Zoology, The Hebrew University of Jerusalem, Israel Vanda Reich. Department of Zoology, The Hebrew University of Jerusalem, Israel Norman Rusft{orth, Department of Biology, Case ·Western Reserve University, Cleveland, Ohio 44106 Charles L. Rutherford, Biology Department, Virginia Polytechnic Institute and State University, Blacksburg, Virginia 24061 H. Chica Schaller, Max-Planck-Institut fUr medizinsche Forschung, Abteilung Biophysik, 6900 Heidelberg, Federal Republic of Germany Tobias Schmidt, Max-Pianck-Institut fiir medizinsche Forschung, Abteilung Biophysik, 6900 Heidelberg, Federal Republic of Germany G. Scott Smith, Developmental Biology Center and Department of Developmental and Cell Biology, University of California, Irvine, California 9271 7 Alan E. Stiven, Department of Biology 046A, University of North Carolina, Chapel Hill, North Carolina 27514 Tsutomu Sug~vama, National Institute of Genetics, Mishima, Shizuoka-ken 411, Japan viii Contributors Joseph R. Voland, Department of Pathology. University of California, San Diego, La Jolla, California Nancy Wanek, Developmental Biology Center and Department of Developmental and Cell Biology. University of California, Irvine, California 92717. Current address: Department of Biology and Health Science, Chapman College, Orange, California 92666. Vaman S. Waravdekar, Microbiological Associates, Bethesda, Maryland 20016 Richard L. JVood, Department of Anatomy, University of Southern California, School of Medicine, Los Angeles, California 90007 William Farnsworth Loomis (1914-1973) The fall of 197 3 saw the passing of a unique scientist and individual who more than anyone was responsible for the current renaissance in the use of hydra as an experimental animal in biological research; he introduced the precision, quantification, and methodology of the physical sciences to the study of this simple invertebrate. I refer, of course, to William Farnsworth Loomis, "Farnie" as he liked to be called by his friends-scientist, explorer, philanthropist, and philosopher. It is fitting that this book be dedicated to him, for Loomis believed that the large advances in biology had their origin with the concomitant development of quantitative methods. ix X William Farnsworth Loomis For example, when he was demonstrating for the first time that mitochondrial particles (called "cyclophorase" then) respire (Green et at., 1948) and the dinitrophenol uncoupled oxidation from phosphorylation (Loomis and Lipmann, 1948), he developed a quick and simple way to calibrate the Warburg respirometer (Loomis, 1949), the major apparatus used in those classic studies. In his research with hydra he is best known for developing methods for culturing the animals en masse, methods which opened the door wide for future generations of biologists to have an ample and continuous supply of this animal in their laboratories. But Loomis did more than simply develop a method for growing the animal; he developed a simple, reliable, repeatable assay for quantifying the logarithmic growth rate of growing cultures of hydra (Loomis, 1954 ). This assay allowed him to determine the extent to which such physicochemical factors as pH, oxygen tension, and ionic strength affected their rate of growth. As a last example of his respect for good methods, when he was concerned about whether the partial pressure of either oxygen or carbon dioxide affected the differentia­ tion of hydra, he developed sensitive micromethods for measuring small amounts of these two gases in fresh water (Loomis, 1954, 1956, 1958). In the ensuing pages I will try to paint a portrait of the man, drawing mostly from recollections of my experiences and conversations with him and also from a few of our letters. I first heard him speak in the winter of 19 53 in a seminar at Johns Hopkins University. I was a graduate student in biochemistry at the time, working in the laboratory of Professor Nathan 0. Kaplan. Loomis, a friend of Kaplan while both were postdoctoral students in the laboratory of Nobel Laureate Fritz Lipmann at Massachusetts General Hospital, was invited to Johns Hopkins to present the first seminar on his new research-the developmental biology of hydra. I remember so well his first slide, a homemade one impressed on cellophane from carbon paper. It was a drawing of a coin with arrows to the side indicating that when considering any problem, "there are two sides to every coin." He was concerned with the problem of cancer-which, according to his son, Pro­ fessor William F. Loomis, Jr., of the University of California, San Diego, was his long-term interest. On one side of the coin were listed properties of normally developing cells and on the other, properties of cancerous cells. Loomis believed that to understand abnormal development we first had to be firmly grounded in knowledge of the development and differentiation of normal cells. His second slide was that of a hydra, because he said that in order to study development he wanted to examine a simple biological system. Not being trained in invertebrate biology, Loomis thumbed through Ralph Buchsbaum's classic Animals Without Backbones ( 1948), played around with the idea of using William Farnsworth Loomis xi sponges, only to find them too difficult to work with, and settled upon the next example of lower invertebrates described by Buchsbaum-hydra. Most of us know the scientific works of Loomis, but few know the man. What kind of person