Table Of ContentSpringer Series in
Experimental Entomology
Thomas A. Miller, Editor
Springer Series In Experimental Entomology
Editor: T. A. Miller
Insect Neurophysiological Techniques
By T.A. Miller
Neurohormonal Techniques in Insects
Edited by T.A. Miller
Sampling Methods in Soybean Entomology
By M. Kogan and D. Herzog
Neuroanatomical Techniques
Edited by N.J. Strausfeld and T.A. Miller
Cuticle Techniques in Arthropods
Edited by T.A. Miller
Measurement of Ion Transport and Metabolic Rate in Insects
Edited by T.J. Bradley and T.A. Miller
Techniques in Pheromone Research
Edited by H.E. Hummel and T.A. Miller
Functional Neuroanatomy
Edited by N.J. Strausfeld
Measurement of
Ion lransport and
Metabolic Rate
in Insects
Edited by
Timothy J. Bradley
Thomas A. Miller
With Contributions by
J.H. Anstee· K. Bowler· T.J. Bradley
D. Brandys· J.W. Hanrahan· A.A. Heusner
J. Machin· S~H.P. Maddrell· L.J. Mandel
J. Meredith· T.A. Miller· M.J. O'Donnell
J.E. Phillips • K. Shima· M.L. Tracy
With 59 Figures
Springer-Verlag
New York Berlin Heidelberg Tokyo
Timothy J. Bradley Thomas A. Miller
Department of Developmental Department of Entomology
and Cell Biology University of California
School of Biological Sciences Riverside, California 92521
University of California USA
Irvine, California 92717
USA
Library of Congress Cataloging in Publication Data
Main entry under title:
Measurement of ion transport and metabolic rate in
insects.
Bibliography: p.
Includes index.
1. Insects-Physiology. 2. Biological transport.
3. Epithelium. 4. Entomology-Methodology. I. Miller,
Thomas A. II. Bradley, T. J. (Timothy J.)
QL495.M4 1983 595.7'01858 83-6683
© 1984 by Springer-Verlag New York Inc.
Softcover reprint of the hardcover 1st edition 1984
All rights reserved. No part of this book may be translated or reproduced in
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The use of general descriptive names, trade names, trademarks, etc., in this
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Typeset by MS Associates, Champaign, Illinois.
987 6 5 4 3 2 1
ISBN -13 :978-1-4613-8241-6 e-ISBN -13 :978-1-4613-8239-3
DOl: 10.1007/978-1-4613-8239-3
Series Preface
Insects as a group occupy a middle ground in the biosphere between bacteria and
viruses at one extreme, amphibians and mammals at the other. The size and general
nature of insects present special problems to the student of entomology. For
example, many commercially available instruments are geared to measure in
grams, while the forces commonly encountered in stUdying insects are in the
milligram range. Therefore, techniques developed in the study of insects or in
those fields concerned with the control of insect pests are often unique.
Methods for measuring things are common to all sciences. Advances sometimes
depend more on how something was done than on what was measured; indeed a
given field often progresses from one technique to another as new methods are
discovered, developed, and modified. Just as often, some of these techniques find
their way into the classroom when the problems involved have been sufficiently
ironed out to permit students to master the manipulations in a few laboratory
periods.
Many specialized techniques are confined to one specific research laboratory.
Although methods may be considered commonplace where they are used, in
another context even the simplest procedures may save considerable time. It is
the purpose of this series (1) to report new developments in methodology, (2) to
reveal sources of groups who have dealt with and solved particular entomological
problems, and (3) to describe experiments which may be applicable for use in
biology laboratory courses.
THOMAS A. MILLER
Series Editor
Contents
1. Introduction
T.J. BRADLEY and T.A. MILLER.................................... 1
2. In Vitro Techniques for Studies of Malpighian Tubules
M.J. O'DONNELL and S.H.P. MADDRELL
With 3 Figures . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5
3. Methods for the Study of Transport and Control in Insect Hindgut
J.W. HANRAHAN, J. MEREDITH, J.E. PHILLIPS, and D. BRANDYS
With 8 Figures .. . .. . .. . .. . .. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 19
4. The Study of Atmospheric Water Absorption
J. MACHIN
With 11 Figures... . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 69
5. Microrespirometry in Small Tissues and Organs
K. SLAMA
With 6 Figures. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. .. 10 1
6. Spectrophotometry and Fluorometry in Ion Transport Epithelia
LAZARO J. MANDEL
With 14 Figures. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 131
VIII Contents
7. Coulometric Measurement of Oxygen Consumption in Insects
A.A. HEUSNER and M.L. TRACY
With 11 Figures.. .. .. . .. . .. .. ... . .. ... ... . .. .. .. . .. ... .. .. . .. . .. . ... 163
8. Techniques for Studying Na+, K+ -ATPase
J.H. ANsTEE and K. BOWLER
With 6 Figures ...................................................... 187
Index ..................................................................... 221
Contributors
J.H. ANSTEE
Department of Zoology, University of Durham, Durham DHI 3LE, England
K. BOWLER
Department of Zoology, University of Durham, Durham DHI 3LE, England
T.J. BRADLEY
Department of Developmental and Cell Biology, School of Biological Sciences,
University of California, Irvine, California 92717, USA
D. BRANDYS
Department of Zoology, University of British Columbia, Vancouver, British Co-
lumbia V6T lW5, Canada
J. W. HANRAHAN
Department of Zoology, University of British Columbia, Vancouver, British Co-
lumbia V6T lW5, Canada
A.A. HEUSNER
Department of Physiological Sciences, School of Veterinary Medicine, University
of California, Davis, California 95616, USA
J. MACHIN
Department of Zoology, University of Toronto, Toronto, Ontario M55 lAl,
Canada
x Contributors
S.H.P. MADDRELL
Department of Zoology, University of Cambridge, Cambridge CB2 3EJ, England
LAZARO J. MANDEL
Department of Physiology, Duke University Medical Center, Durham, North
Carolina 27710, USA
J. MEREDITH
Department of Zoology, University of British Columbia, Vancouver, British Co-
lumbia V6T lW5, Canada
T.A. MILLER
Department of Entomology, University of California, Riverside, California 92521,
USA
M.J. O'DONNELL
Department of Zoology, University of Cambridge, Cambridge CB2 3EJ, England
J.E. PHILLIPS
Department of Zoology, University of British Columbia, Vancouver, British Co-
lumbia V6T lW5, Canada
K. SLAMA
Department ofInsect Physiology, Institute of Entomology CSA V, Praha 6, Czech-
oslovakia
M.L. TRACY
Department of Physiological Sciences, School of Veterinary Medicine, University
of California, Davis, California 95616, USA
Chapter 1
Introduction
T. J. Bradley and T. A. Miller
Arthropods, particularly the insects, offer distinct advantages as sources of ex·
perimental material for the study of ion and fluid transport. Insect epithelia
are histologically simple. They are one cell layer thick and generally not asso·
ciated with underlying cellular connective tissues. These characteristics have
contributed to the substantial successes to date in isolating intact and viable
epithelial organs for study in vitro. The chapters in this volume by O'Donnell
and Maddrell (Chapter 2) and by Hanrahan et al. (Chapter 3) demonstrate the
elegant techniques that have been developed to exploit these advantages. Insect
physiologists have pioneered many miniaturizations of analytical techniques in
response to the challenges of working with small animals. Machin (Chapter 4)
describes in detail a number of the techniques that he has developed to analyze
the uptake of water from subsaturated air by small arthropods.
In other areas of ion transport physiology, our information on insect tissues
has lagged behind that of workers using other organisms. One clear example of
this is in the area of the biochemistry of ion·transporting enzyme complexes.
The difficulty of obtaining large quantities of isolated, homogeneous insect
tissue has complicated the isolation and characterization of the metabolic units
responsible for ion translocation. One notable exception to this has been the
work on Na+, K+-ATPase in the insects, a topic reviewed by Anstee and Bowler
(Chapter 8).
The difficulty of investigating insect transport systems through biochemical
analysis has led some physiologists to examine the links between transport and
cell metabolism. One of the attractions of ion and water transport as physiologi-
cal phenomena for study is the capacity to place these events in a physical con-
text; that is, ion gradients and osmotic gradients can be equated with units of
