Advances in Mutagenesis Research

224 Pages·1990·5.462 MB·English

by  H. Hayatsu (auth.), Professor Dr. Günter Obe (eds.)

1
Advances
in Mutagenesis Research _____
Editor-in-Chief
G. Obe, Essen
Editorial Board
H.J. Evans, Edinburgh
A.T. Natarajan, Leiden
H.S. Rosenkranz, Cleveland
F.H. Sobels, Leiden
T. Sugimura, Tokyo
Advances
in Mutagenesis
Research 1
Edited by G. Obe
With Contributions by
M. Bauchinger F.K. Ennever H. Hayatsu
R. Huber C. Kessler I. Mellon H. Nathel
C.A. Smith R.M. Speed
With 63 Figures
Springer-Verlag Berlin Heidelberg New York
London Paris Tokyo Hong Kong
Professor Dr. GONTER 0BE
FB9 der Universitiit
Gesamthochschule Essen
UniversitiitsstraBe 5
Postfach 103764
4300 Essen 1, FRG
ISBN-13:978-3-642-74957-5 e-ISBN-13 :978-3-642-74955-1
DOl: 10.1007/978-3-642-74955-1
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© Springer-Verlag Berlin Heidelberg 1990
Softcover reprint of the hardcover 1st edition 1990
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2131/3145-543210 - Printed on acid-free paper
Foreword to the Series
Mutations are permanent changes in the genetic material. These
changes can comprise single genes (gene mutations), the structure of
the chromosomes (chromosome mutations), or the number of the
chromosomes (genome mutations). Since H.J. Muller presented his
paper The problem of genic modification at the 5th International
Congress of Genetics in Berlin on the 15th of September, 1927, in
which he brilliantly showed that X-rays induce mutations in the fruit
fly Drosophila, we have learnt that a plethora of agents, including
ionizing and nonionizing radiations, chemicals, and viruses, can in
duce mutations. In most of the cases, induced mutations are delete
rious to the cells or the organisms in which they occur, and we cannot
justify damaging the genetic material of organisms, including oursel
ves, by introducing man-made mutagenic agents into the environ
ment. To prevent this, chemicals must be tested for their possible
mutagenicity in a variety of test systems before they can be used. This
has opened a field of applied genetic research, namely, genetic toxi
cology. Comparative analyses led to the concept that mutagenic
agents can be expected to be also carcinogenic. The theory of the
origin of cancer by mutations has gained experimental proof by the
finding that oncogenes, when changed by mutations, can give rise to
cancer.
Basic research in the field of mutation research has unraveled
some of the molecular mechanisms underlying the origin of mutations
and the complex reaction of cells to induced changes in their DNA.
These cellular reactions can eventually lead to the restoration of the
original structure of the DNA, but, via misrepair, can also give rise
to mutations.
There are still many open questions. The molecular mechanisms
leading to mutations are only partially known. In view of the fact that
about 6 in 1000 newborn children have a chromosomal alteration, it
would be especially important to understand how chromosome and
genome mutations are produced.
Molecular changes in the DNA and the reaction of the cells to
such changes result in typical mutation rates which reflect the evo
lutionary history of the organisms in question. Mutations are one of
the sources of variability which is the prerequisite for natural selection
and for evolution; but since mutations can also result in various
VI Foreword to the Series
deleterious effects, such as hereditary diseases, a population can only
survive when the mutation rates are not too high and not too low,
i.e., mutation rates are delicately balanced. Elevations of the muta
tion rates would have considerable consequences. It would lead to an
increase in the frequencies of cancers and would represent a great
risk for the evolutionary future of a species; a scenario in which
humans are fully included.
In view of these implications, mutation research has two aims:
1. To understand the molecular mechanisms leading to mutations
and
2. to prevent a thoughtless introduction of mutagenic agents into our
environment.
Both aspects, namely basic and applied ones, will be treated in
the new series Advances in Mutagenesis Research. The articles will
deal with current developments in the field of mutation research and
will help the reader to orient himself in this centrally important area
of biology.
Prof. Dr. GUNTER OBE
Contents
Blue Cotton - Broad Possibility in Assessing Mutagens/Carcinogens
in the Environment
H. HAYATSU (With 9 Figures) ................................... 1
1 Introduction .................................................. 1
2 Preparation and Properties of Blue Cotton ................... 5
3 Specificity as a Mutagen Adsorbent .......................... 7
4 Use of Blue Cotton in Studies of Mutagens/Carcinogens ..... 13
5 Other Applications ........................................... 21
6 Conclusion and Perspectives .................................. 22
References ....................................................... 23
Meiosis in Mammals and Man
R. M. SPEED (With 10 Figures) ................................. 27
1 Introduction .................................................. 27
2 Cytological Technology from Squash to Surface Spreading ... 31
3 Mutation and Meiosis ......................................... 38
4 Non-Disjunction and Aneuploidy. . . . . . . . .. . . . . . . . . . . . . . . . . . .. 56
5 Future Molecular Approaches to Meiosis ..................... 59
References ...................................................... 61
Mutagen.Mutation Equilibria in Evolution
H. N01HEL (With 7 Figures) .................................... 70
1 Introduction .................................................. 70
2 Phylogeny of Irradiated RO-Populations of Drosophila
melanogaster and Their Adaptations to X-Rays .............. 71
3 Adaptive Resistance of RO-Populations to the Induction
of Genetic Radiation Damage and Genetic Factors
Controlling It ................................................. 76
4 The Role of Transposons in Relative Radioresistance of
RO-Populations and in Maintaining Mutagen-Mutation
Equilibria ..................................................... 79
5 ENU-Treated Drosophila Populations and the Role of Sterility
in Mutagen-Mutation Equilibria .............................. 82
6 Conclusion .................................................... 85
References ...................................................... 86
VIII Contents
Development and Perspectives of the Human Lymphocyte
Micronucleus Assay
R. HUBER and M. BAUCHINGER (With 7 Figures) .............. 89
1 Introduction .................................................. 89
2 Lymphocyte Preparation with Destroyed Cytoplasm ......... 90
3 Lymphocyte Preparation with Preserved Cytoplasm .......... 92
4 Modified Harlequin Staining Procedure ...................... 95
5 Autoradiographic Method .................................... 96
6 Cytokinesis-Block Method. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 97
7 Perspectives ................................................... 100
References ...................................................... 102
Detection of Nucleic Acids by Enzyme-Linked Immuno-Sorbent
Assay (ELISA) Technique: An Example for the Development
of a Novel Nonradioactive Labeling and Detection System with
High Sensitivity
C. KESSLER (With 27 Figures) .................................. 105
1 Previous Nonradioactive Nucleic Acid Detection Systems .... 105
2 Highly Sensitive DNA Labeling and Detection System Based
on Digoxigenin: Anti-Digoxigenin ELISA Technique ........ 113
3 Sensitivity, Specificity and Low Background of the Digoxigenin
System ........................................................ 118
4 Optimizing the Individual Steps of the Digoxigenin System ... 121
5 Application of the Digoxigenin System in Various Techniques
Used in Molecular Biology and Medicine ..................... 129
6 Summery and Perspective .................................... 140
References ...................................................... 142
Clues to the Organization of DNA Repair Systems Gained
from Studies of Intragenomic Repair Heterogeneity
C. A. SMITH and I. MELLON (With 3 Figures) .................. 153
1 Introduction .................................................. 153
2 Preferential Repair of Pyrimidine Dimers .................... 155
3 Preferential Repair: Relationship to Transcription ........... 163
4 Repair in Mutant Cells ....................................... 170
5 Repair of Other Lesions ...................................... 176
6 Mutagenesis .................................................. 184
7 Conclusions and Perspectives ................................. 186
References ...................................................... 189
Contents IX
The Use of Short· Term Genotoxicity Tests in Risk Assessment
F. K. ENNEVER ................................................. 195
1 Introduction .................................................. 195
2 Applications of Short-Term Tests ............................. 196
3 Conclusions ................................................... 204
References ...................................................... 205
Subject Index ................................................... 209
Blue Cotton - Broad Possibility in Assessing
Mutagens/Carcinogens in the Environment
H. HAYATSU
Contents
1 Introduction. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1
1.1 What is Blue Cotton? ............... .................. ..................... 2
1.2 Historical Background ..................................................... 3
2 Preparation and Properties of Blue Cotton ................................... 5
2.1 Preparation . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5
2.2 Properties ................................................................ 6
3 Specificity as a Mutagen Adsorbent .......................................... 7
3.1 Classification of Adsorbable and Nonadsorbable Compounds................... 7
3.2 Mechanism of Adsorption............... ....... .......................... .. 10
4 Use of Blue Cotton in Studies of Mutagens/Carcinogens. . . . . . . . . . . . . . .. . . . . . . . . 13
4.1 Mutagens in Food..... ............. ..................... .................. 14
4.2 Cigarette Smoke, and Opium Pyrolysate ..................................... 16
4.3 Urines and Feces ......................................................... 18
4.4 Body Fluids and Tissues ................................................... 20
4.5 Water and Air ............................................................ 20
4.6 Others ................................................................... 21
5 Other Applications ........................................................ 21
6 Conclusion and Perspectives ................................................ 22
References ................ . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 23
Abbreviations
Trp-P-l, 3-amino-l ,4-dimethyl-5H-pyrido[4,3-b ]indole; Trp-P-2, 3-amino-l-methyl-5H-pyr
idol 4,3-b ]indole; Glu-P-l, 2-amino-6-methyldipyrido[1 ,2-a:3' ,2' -djimidazole; Glu-P-2, 2-amino
dipyrido[1,2-a:3',2'-djimidazole; AaC, 2-amino-9H-pyrido[2,3-b]indole: MeAaC, 2-amino-3-
methyl-9H-pyrido[2,3-bJindole; IQ, 2-amino-3-methylimidazo[4,5-j]quinoline: MeIQ, 2-amino-
3,4-dimethylimidazo[4,5-j]quinoline; MeIQx, 2-amino-3,8-dimethylimidazo[4,5-j]quinoxaline;
4,8-Me2IQx, 2-amino-3,4,8-trimethylimidazo[ 4,5-j]quinoxaline; 7,8-Me2IQx, 2-amino-3,7,8-tri
methylimidazo[4,5-j]quinoxaline; PhIP, 2-amino-l-methyl-6-phenylimidazo[ 4,5-b ]pyridine;
MNU, N-methyl-N-nitrosourea.
1 Introduction
The scope of studies on mutagens widened greatly when the notion that carcino
gens overlap with mutagens was introduced by Ames et al. (1973). The mutagen
detection has become increasingly important since the discovery of highly potent
1 Faculty of Pharmaceutical Sciences, Okayama University, Tsushima, Okayama 700, Japan
2 H. Hayatsu
mutagens in cooked foods and subsequent demonstration of their carcinogenicity
in rodents (Sugimura 1988). It seems now that our environment contains a great
number of mutagens, and humans live surrounded by mutagens/carcinogens
(Sugimura 1982; Ames 1983; Ames et al. 1987). As a result, the importance of
assessing the environmental mutagens both qualitatively and quantitatively has
become realized by researchers worldwide, and great efforts have been made to
accomplish this task. A difficulty one encounters in attempting this assessment is
that mutagens, either those of known structures or of unknown nature, are present
only in tiny amounts in the environmental materials. Furthermore, these materials
are almost infinite in their kinds and numbers. Consequently, methodological
advancement has been, and is, very important for the development of this area
of science. An efficient means to detect mutagens in complex mixtures would
undoubtedly facilitate the progress of the study.
The blue cotton method is one such means, recently introduced by our own
research group. This method has been used extensively for isolating mutagens
and for detecting mutagenicity in the environment. In this chapter, I would like
to summarize the present status of this new methodology.
1.1 What is Blue Cotton?
Blue cotton is cotton bearing covalently linked blue pigment, copper pathalocy
anine trisulfonate (Hayatsu et al. 1983a). The linkage connecting the pigment to
cellulose is illustrated in Fig. 1. Figure 2 shows the picture of blue cotton, together
with that of blue rayon, which is a recently developed, modified version of blue
cotton. The synthesis of blue cotton can be done by using a simple one-step
reaction shown in Fig. 3.
The usefulness of blue cotton lies in its unique property to adsorb aromatic
compounds having three or greater numbers of fused rings. The adsorption ca
pacity is strong, and the selectivity for adsorbing this class of compound is high.
The fact that many of such polycyclic aromatic compounds in the environment
are mutagenic and often carcinogenic makes blue cotton extensively useful as a
means to efficiently purify those mutagens from crude samples.
Blue Cotton Fig. 1. Structure of blue cotton