Table Of ContentT E C H N I Q U ES IN
E N D O C R I NE RESEARCH
PROCEEDINGS OF A NATO ADVANCED
STUDY INSTITUTE HELD AT
STRATFORD-UPON-AVON, ENGLAND,
UNDER THE AUSPICES OF
THE UNIVERSITY OF BIRMINGHAM
Edited by
PETER ECKSTEIN and FRANCIS KNOWLES
Department of Anatomy, University of Birmingham
1963
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LIST OF PARTICIPANTS
Director
SIR FRANCIS KNOWLES,* University of Birmingham, Department of Anatomy,
Birmingham, England
Secretary
W. P. DALLAS ROSS, University of Birmingham, Department of Anatomy,
Birmingham, England
AMOROSO, E. C.,* Royal Veterinary College, University of London, England
ANTONIADES, H. N.,* Blood Characterization and Preservation Laboratory,
Protein Foundation Inc., Massachusetts, U.S.A.
BARER, R.,* Department of Human Anatomy, University of Oxford, England
BARKER, S. Α., University of Birmingham, Department of Chemistry, England
BARONDES, S., Laboratoire d'Enzymohgie, C.N.R.S., Gif-sur-Yvette, France
BEAUMONT, HEATHER M., Department of Anatomy, University of Birming-
ham, England
BLOOM, G. D. Institutionen for Cellforskning, Karolinska Institutet, Stock-
holm, Sweden
BROWN-GRANT, K., Department of Physiology, University of Birmingham,
England
BUSH, I. E.,* Department of Physiology, University of Birmingham, England
BUTT, W. R., Department of Clinical Endocrinology of the Birmingham and
Midland Hospital for Women, University of Birmingham, England
CROOKE, A. C, Department of Clinical Endocrinology of the Birmingham and
Midland Hospital for Women, University of Birmingham, England
DEANESLY, RUTH, School of Agriculture, University of Cambridge, England
DE DUVE, C.,* Universite de Louvain, Laboratoire de Chimie Physiologique,
Belgium
DELOST, P., Laboratoire de Physiologie Animale, Université de Clermont,
France
DIETERLEN, FRANÇOISE, Laboratoire d'Embryologie Expérimentale, Collège de
France, logent-sur-Marne, France
DODD, J. M.,* Department of Zoology, University of Leeds, England
ECKSTEIN, P., Department of Anatomy, University of Birmingham, England
EULER, U. S. VON,* Fysiologiska Institutionen, Karolinska Institutet, Stock-
holm, Sweden
FICQ, ADRIENNE,* Faculté des Sciences, Université Libre de Bruxelles, Belgium
FINE AN, J. B., Department of Medical Biochemistry and Pharmacology,
University of Birmingham, England
* Participants who contributed papers
ν
Vi LIST OF PARTICIPANTS
FLAMENT, JACQUELINE,* Neuropathology Department, University Hospital
Brugmann, Brussels, Belgium
FRANCHI, L., Department of Anatomy, University of Birmingham, England
GELL, P. G. H., Department of Experimental Pathology, University of Bir-
mingham, England
GEMZELL, C.,* Akademiska Sjukhuset, Uppsala, Sweden
GILBERT, G. Α., Department of Chemistry, University of Birmingham,
England
GLASS, Α., Department of Anatomy, University of Birmingham, England
GREEN, S. H., Department of Anatomy, University of Birmingham, England
HAFFEN, KATY, Laboratoire d'Embryologie Expérimentale, Collège de France,
Nogent-sur-Marne, France
HALL, KATHLEEN, Department of Physiology, University of Birmingham,
England
HELLER, H., Department of Pharmacology, University of Bristol, England
HICKMAN, J. Α., Department of Anatomy, University of Birmingham, England
HICKS, R. MARION, Bland-Button Institute of Pathology, Middlesex Hospital,
London, England
HODGKIN, DOROTHY, M.C.,* Chemical Crystallography Laboratory, University
of Oxford, England
HOLMES, R. L.,* Department of Anatomy, University of Birmingham, England
HOLT, S. J.,* Courtauld Institute of Biochemistry, Middlesex Hospital
Medical School, London, England
HUNTER, SHEILA, Department of Physiology, University of Birmingham,
England
JONES, A. S., Department of Chemistry, University of Birmingham,
England
KARLSON, P.,* Physiologisch-Chemisches Institut der Universität München,
Germany
KEIL, BARBARA, Physiologisch-Chemisches Institut der Universität München,
Germany
KLEIN, M.,* Institut de Biologie Médicale, Université de Strasbourg, France
KNIGHTS, Β. Α., Department of Anatomy, University of Birmingham, England
KOVACIC, NADA, Department of Clinical Endocrinology of the Birmingham
and Midland Hospital for Women, University of Birmingham, England
KRAICER, P., The Weizmann Institute of Science, Rehovoth, Israel
KROHN, P. L.,* Department of Anatomy, University of Birmingham, England
LANE-PETTER, W.,* M.R.C. Laboratory Animals Centre, Carshalton, England
LEDERER, E.,* Laboratoire de Chimie Biologique, Faculté des Sciences,
Université de Paris, France
MANDL, ANITA M., Department of Anatomy, University of Birmingham,
England
MARSHALL, W. Α., Department of Anatomy, University of Birmingham,
England
MAYER, G.,* Laboratoire d'Histologie, Faculté de Médecine, Bordeaux, France
MEUNIER, J. M., Ecole National de Médecine et de Pharmacie de Poitiers,
France
LIST OF PARTICIPANTS VÜ
PARKES, A. S.,* Physiological Laboratory, University of Cambridge, England
PENNELL, R. B.,* Blood Characterization and Preservation Laboratory, Protein
Foundation Inc., Massachusetts, U.S.A.
PEPE, F. A.,* Department of Anatomy, University of Pennsylvania, Phila-
delphia, U.S.A.
PEYRE, Α., Institut de Physiologie, Université de Toulouse, France
RAYNAUD, A.,* Institut Pasteur, Laboratoire de Sannois, France
ROGERS, A. W., Department of Anatomy, University of Birmingham, England
SALZGEBER, BERTHE, Laboratoire d'Embryologie Experimentale, Collège de
France, Nogent-sur-Marne. France
SCHEIB, DENISE, Laboratoire d'Embryologie Expérimentale, Collège de France,
Paris, France
SEKERIS, CE., Physiologisch-Chemisches Institut der Universität München,
Germany
SHELESNYAK, Μ. Ο.,* Department of Experimental Biology, Weizmann Institute
of Science, Rehovoth, Israel
THOMAS, G. H., Department of Anatomy, University of Birmingham, England
TOMKINS, G. M.,* Laboratoire d'Enzymologie, C.N.R.S., Gif-sur- Yvette, France
WAKELING, Α., Department of Anatomy, University of Birmingham, England
WALL, PATRICIA, Department of Physiology, University of Birmingham,
Englxind
WALTON, K. W., Department of Experimental Pathology, University of
Birmingham, England
WHITE, R. G.,* Department of Bacteriology, London Hospital Medical College,
London, Engkind
WOLF, ANNELIESE, Department of Clinical Endocrinology of the Birmingham
and Midland Hospital for Women, University of Birmingham, England
ZUCKERMAN, SIR SOLLY, Department of Anatomy, University of Birmingham,
England
Sir Robert Althen, Vice Chancellor of the University of Birmingham, under
whose auspices the Conference was held, welcomed the participants and
addressed them on the Epistemology of Endocrinology
FOREWORD
by
PROFESSOR SIR SOLLY ZUCKERMAN
A NATO Science Committee was established in 1958 under the
chairmanship of a full-time Science Adviser, now the Assistant Secre-
tary-General for Scientific Affairs, in order to help implement a decision
taken in the preceding year by the Heads of Government that the
scientific and technical resources of the Western Alliance should be
strengthened. The Committee has done much to promote scientific
co-operation between the dînèrent member nations, and in particular
has supported various projects designed to bring able young scientists
into contact with more mature workers in various fields of research.
A Ν A TO Science Fellowship Programme has enabled many hundreds of
students to study in countries other than their own ; a Research Grants
Programme has supported research projects, both national and inter-
national in character ; and an Advanced Study Institute Programme has
made it possible to arrange meetings at which scientists of world-wide
reputation discuss current problems with younger people.
These Institutes differ from more formal scientific conferences, in
so far as they provide an opportunity for unhurried discussion in
relatively small groups between workers of different nationalities and
different levels of attainment. In this way young scientists are stimu-
lated by a high level of instruction and discourse, while more senior
scientists have an opportunity to engage in a frank exchange of views.
The success of these Study Institutes, a field in which NATO has
taken the lead in the West, may be gauged from the increasing finan-
cial support they have received. Starting with 100 000 dollars in 1959,
their budget in 1961 amounted to 300 000 dollars. Nationals of forty
countries participated in the Study Institutes held in I960.
Institutes in physics predominated at the start, but more recently a
number of Advanced Study Institutes in biological fields have been
arranged. One of the first of these has resulted in the present volume of
published papers. The organizers of this meeting, held at Stratford-
upon-Avon, felt that the very rapid advances in analytical techniques
during recent years, and the growing diversity of the subject, made it
desirable to restate some of the basic problems of endocrinology. Those
who attended the conference will not forget the unique opportunities
ix
χ FOREWORD
which it presented for a detailed exchange of views between workers
in disciplines which were very different yet all relevant to research in
endocrinology. The meeting marked a milestone in the progress of the
subject and I am delighted to see that most of the important papers
which were read at this meeting are now available to all research
workers.
INTRODUCTION
by
SIR FRANCIS KNOWLES
The NATO Advanced Study Institute, "Techniques in Endocrine
Research", held at Stratford-upon-Avon in September 1962, brought
together workers trained in different disciplines who, though pursuing
closely related fields of study normally have little opportunity of meet-
ing for constructive discussion. Two major groups were represented :
first, scientists who employ already established or recently developed
techniques in their researches into endocrinology, and, second, chemists,
physicists and others who have developed or are developing techniques
which potentially have a value in endocrinology. The papers presented
at the Study Institute varied in title and content, but together provided
a central theme for discussion, namely an evaluation of the techniques
used in endocrine research, their opportunities and their limitations.
Developments in physics and chemistry during the past thirty years
have revolutionized biological research. Refinements of biochemical
analysis have made possible the separation of minute quantities of
chemical substances and their identification in the blood stream.
Workers using X-ray crystallography have unravelled complex
patterns of molecular structure. Histochemistry and autoradiography
have permitted the identification of hormones and associated com-
pounds in tissues, and have indicated modes of synthesis and the
metabolic pathways of hormones. A greater understanding of antibody
formation has made possible new techniques applicable to endo-
crinology. The development of the electron microscope and other
forms of microscopy has led to a better understanding of the fine
structure of endocrine tissues. The refinement of these and other
techniques has in sum resulted in an impressive accumulation of data
on the structure and function of endocrine systems. Yet it is pertinent
to enquire whether the application of these many and varied techniques
has given us a true picture of such a complex, integrated and balanced
system as the endocrine system.
In his introductory address Professor Amoroso draws attention to
the fact that the adrenal cortex has, in the hands of the biochemists,
yielded an impressive array of active substances ; nonetheless there are
grounds to believe that very few of these in fact represent functional
hormones. Similarly, methods of electrophoresis have made possible
the isolation of a number of substances which on injection bring about
xi
xii INTRODUCTION
pigment movements in the chromatophores of crustaceans; but it is
far from clear whether they represent substances which are involved
in the normal physiology of colour change in these animals. The
biochemical analysis of hormones, especially those in the blood stream,
is an evident pre-requisite to a proper understanding of the nature and
activity of hormones ; yet we must enquire whether available methods
of chemical analysis give us a true account of hormones present in
endocrine organs, or in the blood, or whether the methods used may
themselves contribute to the formation of some of the substances
which they identify.
Similar considerations apply to the evaluation of results obtained
by using electron microscopy and autoradiography, especially in the
study of the endocrine function of certain components of the nervous
system. Some of the results yielded by these methods are equivocal.
For example, the known neurosecretory systems in Arthropods and
Chordates are characterized by the presence of vesicles of a limited
size range, which have been termed 'elementary neurosecretory
vesicles' ; yet similar vesicles are ubiquitous in the nervous systems of
some lower invertebrates, apparently in normal motor neurons.
Autoradiography suggests hormone synthesis is performed in the
perikaryon; yet electron microscopy indicates metabolic transforma-
tions along the course of neurosecretory axons.
Biochemical studies to determine the mode of action of hormones
at the cellular level have not as yet given a clear-cut answer to the
question of the primary mode or site of action of a hormone—on
membranes, enzymes or genes? Evidence for each mode of action
is forthcoming. Possibly different hormones have fundamentally
different actions. Certainly one might expect that a hormone which
influences development might act directly on gene loci, whereas
another with direct action on an effector, such as a chromatophore,
might exert its primary action on membranes or enzyme systems.
Logically a hormone must ultimately affect enzyme systems, since
it is by these that material and energy transformations in the cell are
effected. The interdependence of enzymes and genes and membrane
systems is, however, so intimate that it is not easy to determine on
which a hormone may have its primary effect.
The more we survey the data obtained by the application of refined
physical and chemical techniques to endocrine research, the more we
may be impressed by the fragmentary picture which they provide, and
the difficulties of interpretation in terms of the organic whole. The
success of the early extraction and injection experiments using animals
of different species raised hopes that relatively few chemical compounds
might bring about the various endocrine effects observed in different
INTRODUCTION xiii
animals. Within recent years, however, biochemical studies on MSH
and ACTH have shown not only that the melanocyte-stimulating
hormones of different species may have different molecular structures,
but that the two principles MSH and ACTH may, on account of their
molecular resemblance, exert somewhat similar effects under experi-
mental conditions. As Professor Gemzell has shown, the most effective
and meaningful hormone research and therapy is that which employs
but a single species. We may indeed proceed even further along this
line of thought and enquire whether, as Dr. Lane Petter has sug-
gested, it may not be prudent to pay attention to the possibility of
differences of hormone balance between different strains of the same
species. There is, moreover, the possibility not only that the endocrine
balance of an animal may vary, at different times of year, but even
that significant difference may occur in the same individual at different
times in a 24-hour cycle. Studies on the diurnal rhythms of colour
change and locomotor activity in Arthropods have demonstrated
clearly that the endocrine system in these animals varies in a cyclical
manner, and can be upset if the animals are removed from their
normal environment. We should therefore consider whether an animal
in captivity has an endocrine balance identical to that of its free-living
counterpart.
Animals are organisms, maintaining a delicate state of homeostasis
by intricate regulatory mechanisms of which the endocrine system is
but one. Analytical techniques can, by reason of their nature, describe
only component parts of such a system. In the early days of endo-
crinology the approach was fundamentally a biological one, in which
the behaviour of the animal was studied in relation to endocrine
control, and the results expressed as far as possible in chemical and
physical terms. The emphasis has changed to some extent, and the
problems of endocrinology are often now expressed more in terms of
chemical or physical rather than biological parameters. In consequence,
a dichotomy has arisen and the organizers of the Stratford-upon-Avon
conference felt that it would be useful to re-affirm the basic problems
of endocrinology, and at the same time to give an opportunity for those
who are primarily interested in the biological aspects of endocrinology
and those whose primary interests lie in the development of new
techniques to meet and exchange views.
These published proceedings contain most of the papers that were
read at Stratford. Unfortunately a few speakers did not feel able,
because of pressure of other work, to present written accounts for
publication. This volume is the poorer as a result, but I feel most
grateful to these, as to the other speakers, for the trouble they took
to present communications at the meeting and to make the discussions
