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SYMPOSIUM: CYTOGENETICS AND CELL BIOLOGY

Interrelationships Between Recent Developments in Molecular


Genetics and Cytogenetics and
Animal Breeding 1

N. S. FECHHEIMER
Department of Dairy Science
2027 Coffey Road
The Ohio State University
Columbus 43210

ABSTRACT INTRODUCTION
Animal breeding traditionally has Successful animal breeding schemes are
entailed devising means to apply quanti- based upon two primary components, one
tative and population genetic theory academic and one commercial. The appropriate
to increase productive capacity of live- function of the academic component is explor-
stock. A highly developed and successful ation of new ground, development of novel
industry has been built on foundations ideas and methodologies that enable new
established by academic animal breeders. questions to be asked, and building a store-
Recent developments in related sciences house of pertinent knowledge. The commercial
such as reproductive biology, molecular component adapts appropriate segments of the
biology, cellular biology, and cyto- new knowledge to build a business to accumu-
genetics offer prospects for the emer- late and sell improved germ plasm, in various
gence of a number of methodologies that forms, to animal producers. There occurs very
might usefully be applied to animal little duplication of effort between the two.
breeding. Scientists engaged in develop- Academic animal breeders have concentrated
ment of the newer technologies are not largely on research and monitoring the opera-
wholly familiar with the livestock in- tion of the industry whereas the commercial
dustry, its breeding structure, its ob- enterprises have dealt with development and
jectives, its institutions or its peculiarities. sales.
Animal breeders, however, are not fully This paper relates largely to academic animal
cognizant of the scientific advances being breeding, its present state, and its prospects. It
made in related fields, their potential for reviews briefly the approaches that have been
development and application or their used, their accomplishments, and their con-
limitations, and therefore, animal breeders sequences. The rapid growth of knowledge in
have not seriously thought about how related fields, viz. reproductive, molecular, and
they might be integrated most usefully cell biology and cytogenetics are noted. Means
and efficaciously into the animal breeding for their integration into academic animal
enterprise. A collaboration is needed in breeding are explored. Particular emphasis is
which the laboratory scientists produce given to speculation about modification of
new ideas, products, and methods and the approaches taken to the general objective of
animal breeders-using system analysis, genetic improvement of animals. Questions are
simulation procedures, and laboratory raised as to how developments in the more
animal and livestock breeding tests-help recently developed fields might best be inte-
make rational choices, partially direct grated into animal breeding research programs,
work of the laboratory scientists, help the how the objectives for animal breeding might
industry integrate new methods, and be altered, and how the enterprise might be
monitor the extent of success of adapted organized to enable the most effective use of
innovations. the limited resources available to departments
of animal science.

Received August 9, 1985. C U R R E N T STATE OF


1Salaries and research support provided by State A N I M A L BREEDING
and Federal Funds appropriated to The Ohio Agricul-
tural Research and Development Center, The Ohio The theoretical bases of present day animal
State University. Journal Article Number 152-85. breeding were elaborated by Jay Lush and his

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1744 FECHHEIMER

students 30 to 40 Yr ago and comprise in- breeding research to other programs that are
genious applications from the work on quanti- judged to have greater potential to increase the
tative and population genetics of Fisher, knowledge necessary for even more rapid
Wright, and Haldane. The primary aims are to changes in the genetic complements of livestock
increase the selection differential and the populations.
correlation between the phenotypic evaluations Traditionally trained academic animal breed-
of animals and their true breeding values. ers have made valuable contributions to quan-
Reduction in generation interval enables a titative, population, and developmental genetics.
greater number of genetic alterations of a Much of their effort is devoted to serving as
population per unit of time. Although the consultants to the industry. The very great
theory has remained intact, its application has analytical skills that they possess are being
undergone periodic changes that were enabled employed to monitor and evaluate the system,
by developments in statistics and by technical to deal with periodic difficulties that arise in
innovations. Those advances of greatest benefit the system, and to devise schemes to effect
were the modern computer, which vastly slight alterations in the efficiency of the system.
increased the capability to do large-scale, They quantify, using esoteric statistical proce-
complex computations, and developments in dures, what is already generally known; analyze
reproductive biology, which enabled exploita- skillfully large volumes of performance test
tion of artificial insemination and some asso- data; and train proficient, statistically oriented
ciated technologies. These and other lesser students. It would be beneficial if academic
developments provided the knowledge for animal breeders became increasingly cognizant
establishment of successful animal breeding of the rapid developments in modern biology
industries, which now provide the germ plasm, with a view to incorporate segments of such
especially for producers of dairy and poultry knowledge into the animal breeding enterprise.
products. A brief survey of some of those developments
Commercial enterprises have served producers of greatest promise is made in the following
very well. In North America the artificial section.
insemination organizations have provided for
genetic improvement in milk yield of about
RECENT DEVELOPMENTS WITH
l%/yr (29). Genetic improvement of broiler POTENTIAL APPLICATIONS TO
chickens is even faster. Even in the pig and beef ANIMAL BREEDING
cattle industries, where breeding is not domi-
nated by large breeding companies, application Manipulation of the Reproductive
of sound recommendations by academic animal Process, Gametes, and Zygotes
breeders has enabled influential practicing The technology of artificial insemination has
breeders to make reasonably rapid progress in become the primary method for breeding dairy
characters such as growth rate, efficiency of cattle because it offered such great opportunity
gain, and carcass composition (7). There can be for applying sound genetic principles (12). Its
no doubt that application of animal breeding proper application, commencing in the early
theory yields a predictable, satisfactory rate of 1960's, enabled selection differentials of bulls
improvement. to be vastly increased and enabled accurate
The consequences of the proved effectiveness estimates of breeding values of males for traits
of animal breeding practice are both beneficial expressed only by cows. Universal adoption by
and detrimental. Large commercial establish- the industry of a plan devised by academic
ments, operated by capable, well-trained animal breeders was the basis for an econ-
specialists are serving the industry well. The omically feasible, practical, and successful
breeding industry itself supports, to a small scheme for continued genetic improvement of
extent, some research, which for the most part dairy cattle.
is directed toward solution of short-term But a number of more recently developed or
problems of the industry. Universities, experi- still developing techniques for altering repro-
ment stations, and the United States Depart- duction of livestock have, thus far, received
ment of Agriculture (USDA) are diverting some scant attention by animal breeders. Embryo
of the resources once allocated to animal transfer is being extensively practised in both

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SYMPOSIUM: CYTOGENETICS AND CELL BIOLOGY 1745

beef and dairy cattle industries (23). Coupled uses of the capability for producing chimeras
with superovulation, embryo transfer offers to have been discussed elsewhere (16).
animal breeders a number of opportunities to A further use of micromanipulation is for
enhance the rate of progress in breeding pro- genetic transformation by injection of deoxy-
grams. Genetically superior females can supply ribonucleic acid (DNA) coding for a known
ova for a much larger proportion of replace- gene into a pronucleus of a fertilized ovum. The
ments; monotocous animals could be progeny injected DNA sometimes integrates into one or
tested to increase the accuracy of estimating more chromosomes of the pronucleus and
their breeding values; the generation interval becomes a part of the genome of the embryo
could be reduced by inducing multiple ovula- (18). When such embryos are brought to term,
tion of young females and transferring the reared, and used as breeding animals, some
resulting embryos to mature cows; productivity transmit their new gene to half their progeny,
of beef cows might be increased by supplying indicating that the new gene was fully inte-
each cow with a second embryo, a fair propor- grated into a chromosome of the one-cell
tion of which would survive gestation (24). embryo. This technique enables the transfer
Animals bearing identical genotypes, the of any gene from any source, no matter how
equivalent of naturally occurring monozygotic remote the relationship between donor and
twins, are being produced commerically by recipient, into a population of animals. It offers
splitting early embryos (3). Work is under way great promise as a means to modify function of
to extend the capability, so that greater num- animals for any characteristic or trait that is
bers of identical animals might be produced conditioned to some extent by single genes
(30). It is even possible that some cell type of with major effects.
mature animals possesses totipotency so that
their genotype might be reproduced, yielding
large numbers of young exactly the same as Genetic Analyses
that of the mature animal. During the past 15 yr, startling advances
The genomic content of ova can now be have been made in molecular biology that have
altered by micromanipulation equipment that led to rapid accumulation of knowledge of the
enables nuclei or pronuclei to be withdrawn or genetic complements of higher organisms
inserted into ova (15). These techniques produce including man and mouse. The methodology
polyploid embryos, embryos containing two has yielded surprising new insights into the
haploid genomes from the same parent, or structure and organization of the mammalian
embryos resulting from parents of different genome, its functioning, and its control. Per-
taxa, i.e., wide crosses. Presently these tech- tinent aspects of the techniques of molecular
niques are being used for study of embryological gentics and their applications were reviewed by
phenomena, but they also have potentially Frankham and Gillings (13). Any gene whose
useful application in animal breeding to which products are known, either messenger ribo-
little thought has been given. nucleic acid or protein, can be mapped, isolated,
Micromanipulation techniques are also being or attached to an appropriate vector for intro-
used to produce chimeric embryos either by duction into bacteria or other cells. Following
fusing two embyos to produce an aggregation its amplification in bacteria, multiple copies can
chimera or by injecting one or more totipotent be recovered and analyzed in detail. The
cells into the inner ceil mass of a blastocyst capability is available to reveal the entire
(19). The injected cells might be from another sequence of bases of which individual genes are
embryo or from a cell line derived from a comprised.
teratocarcinoma maintained in cell culture in Elements involved in the control of gene
the laboratory. Frequently the introduced cells function are being identified and manipulated.
become incorporated fully into all tissues of the The DNA segment that controls the gene for
developing chimeric embryo including the germ heavy metal metabolism in mice, the metala-
line. Capabilities made available by use of these thionein gene, was attached to the growth
methods include that of introducing the geno- hormone gene of rats and the entire contruct
type of cells maintained and modified in the transferred, by injection into pronuctei of
laboratory into a breeding population. Other fertilized eggs, into mouse embryos. During the

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postweaning growth period of resulting mice, rapid rate and warrant serious thought, con-
addition of zinc to the diet induced high sideration, and plans for experimental use.
growth hormone production from the liver and
other tissues (18). This experiment illustrates Cell Biology and
the potential to influence productive function Cytogenetics
of animals by altering the copy number of key Advances in methods to establish and
genes and using imaginative ways to control maintain mammalian cell lines in culture and to
their activity in particular organs or tissues at elicit from them knowledge of their genetic
specific times in development. complements have been widely exploited to
Other lines of work have indicated that map a number of human genes. Techniques of
genes affecting a single animal function in some cell genetics enable the localization and study
cases are clustered into short lengths of a of any gene that can be made to function in
chromosome and are n o t randomly located in cultured cells. Thus, the task of gene mapping
the genome (6). Further, when intense selection can proceed rapidly without employing crosses
for resistance to a cytotoxic agent is applied to of classical genetics and observation of variant
cell cultures, the positive response has been phenotypes in embryos or living animals (21).
mediated by localized amplification of a single Cells in culture will tolerate relatively
gene rather than change in frequency of rele- extreme alterations of their genome such as
vant genes at many loci (22). This type of study chromosome segment duplications, deletions,
illustrates the importance of understanding the aneuploidy, and polyploidy. Lines containing
organization of the genome of animals and such aberrations are used to establish the
suggests possible ways in which useful functions location of genes on specific chromosomes by
of animal might be enhanced. deletion and duplication mapping. They are
Recognition of the occurrence of base pair also used to study gene activity in cells bearing
substitutions at fairly high frequencies in the more or fewer than two alleles at a locus. It is a
DNA of animals, including man, was accom- relatively easy matter to produce, isolate, and
plished by analysis of DNA fragments produced propagate cell lines bearing aberrant chromo-
by various restriction enzymes. Application of some complements for a variety of purposes.
the technique to the study of DNA of man has Such cell lines can be studied intensely in
revealed a number of restriction fragment culture and even introduced as component
length polymorphisms. The variant sites exert clones of chimeric embryos. In this way the
no gross phenotypic effects but can be used as effect of a wide array of genome alterations on
markers to map the entire genome (25). Such developing embryos can be observed. This is
silent markers, it is proposed, could then be because when present as only one component
used to indicate the presence or absence of in a chimera, chromosomal alterations that in
other genes or gene clusters to which they pure form are lethal, exhibit attenuated effects
could be shown to be closely linked. (8).
Cells in culture can be transformed, i.e., new The value of cytogenetics to the animal
genes can be introduced into them, by a number breeding enterprise has been well-established
of techniques including direct DNA transfer, (11). Standard and banded karyotypes of all
chromosome-mediated transfer, and viral vec- domesticated food producing animals have been
tors (26). Cell lines can be transformed serially prepared. From these much has been learned of
or otherwise altered genetically to contain a the structure and organization of the chromo-
new, engineered genotype. If the engineering is somes. The importance of chromosomal aber-
done in a culture, the cells of which have rations as a cause of embryonic mortality,
maintained a totipotency, they can subse- stillbirth, gross abnormality, and infertility and
quently be introduced into a breeding popula- sterility of adult animals has been well-estab-
tion by construction of injection chimeras (27). lished in man and mouse. Demonstration of the
This brief discussion of a few of the areas in applicability of much of this knowledge to
molecular genetics presently under study is food-producing animals has accrued slowly but
perhaps sufficient to indicate that techniques is now well under way. Polymorphisms o f
and methods of great potential value to the chromosome structure and banding pattern
animal breeding enterprise are developing at a are ubiquitous, but little is known of the

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phenotypic manifestations of these variants. breeding methods has been slow. Now, however,
Lines of chickens and pigs bearing structural many departments of animal science are adding
rearrangments of chromosome segments have biotechnologists to their faculties, the USDA
been recovered from populations or produced apportioned a sizeable amount of money for
experimentally. Many such lines with marker grants for work in the area, and many spec-
chromosomes are maintained and used to map ulative articles are appearing in popular press.
genes, to establish the parental origin of aber- In spite the emerging awareness of need for
rant gametes, or to produce embryos bearing change, little serious thought has been given to
deletions and duplications of known segments. how the whole array of new methodologies
Cytogeneticists have also contributed use- should be selectively studied, developed, and
fully in collaborative work to map genes by in ultimately exploited by the animal breeding
situ hybridization, to characterize the genome enterprise. With some notable exceptions (17,
of cell lines, to gain a further understanding of 9, 20, 28), animal breeders have hardly begun
the mechanisms of sex determination in man to think about the implications of what is at
and animals, and to diagnose the occurrence of hand. Molecular and cellular biologists are
afflictions caused by chromosome abnormali- trained in fields other than the animal sciences,
ties. Attempts to study the etiology of chromo- and their primary interests are toward pursuit
some abnormalities have led to standard pro- of detailed knowledge for its own sake. Cyto-
cedures for detection of exposure of animals to geneticists, in the absence of a plan for applica-
toxic, mutagenic, and carcinogenic agents, tion of their talents in an integrated scheme for
many of which produce chromosome lesions improvement of animal productivity, have
detectable with cytogenic methods (4). adhered to their own, somewhat independent
agenda (11). Academic animal breeders are
PRESENT STATE OF AFFAIRS bewildered by the flurry of activity in areas
From the brief foregoing summary of devel- they do not understand but which they are told
opments in fields allied to animal breeding it is relate to animal breeding. They have seen their
clear that many powerful new tools to alter the role as one of continuing their traditional and
genetic complement of livestock are becoming familiar activities. It is well that they do,
available for exploration, further refinement, because in spite of great expectations for
and selective application. But molecular biology application of the newer technologies to animal
has changed not only the means for obtaining breeding, such applications are not imminent.
knowledge about the genetic endowment of Meanwhile, the industry must continue to
animals but also the nature of the knowledge function.
one can seek. It was once thought that study of
individual genes and their effects on productive NEED FOR AND EMERGING
NATURE OF COLLABORATIONS
traits of livestock would be useless because it
was impossible using traditional methods to Traditionally trained animal geneticists and
give attention to more than a small fraction of molecular and cellular biologists differ in their
the vast array of genes that influenced perform- base of knowledge, skills, manner of approach
ance of animals for any economically important to problems, and in their understanding of the
trait. Further, if genes with major effects were structure and operations of the livestock
detected it required a long and tedious process breeding industry. They will, however, have a
to transfer them into the populations where common goal, the genetic improvement of
they were desired. The situation is now sub- populations of animals. The presence of all
stantially altered, because large numbers of three types of scientists in departments of
genes can be mapped and isolated from any animal science, with good will and appropriate
species. More than 600 human genes have been leadership, should lead to useful collaborations
mapped, and the pace is accelerating so that which could well enhance the productivity of
almost 150/yr are now being isolated (14). all.
Once useful genes are located, new technologies When they are first brought into depart-
permit their transfer relatively quickly. ments of animal science, molecular biologists
Recognition of the potential use of new are under great pressure to apply their skills to
technology to supplement traditional animal spectacular, but in overall context irrelevant,

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1748 FECHHEIMER

experiments such as production of giant mice played an important role in designing the
or pigs. In the long run their much more useful structures for these industries, their knowledge
role will be in accumulating knowledge of the will be required to help find the optimum
genes involved intimately in influencing pro- strategies and mechanisms for applications of
ductivity of livestock and in gaining an under- newer technologies.
standing of how such genes act and interact; in
learning how many genes affect particular Cytogenetics and Animal
functions, where they are located, how they Breeding
act, what controls their activity, and what the Animal cytogenetics has developed rapidly
interrelationships are among them. While during the past 20 yr and will now make
engaged in this process, molecular biologists important contributions to both the theory and
will also learn a great deal about organization of practice of animal breeding (10, 11). Collabora-
the genome, and its meaning and importance to tions similar to those between molecular
the question of how genetic changes can biologists and animal breeders are needed
be made. Gene duplication and amplification, between each of these two groups and cyto-
directed recombination, nonrandom transmis- geneticists if the capabilities of the latter
sion of alleles, site-specific mutation, and other group are to be incorporated into schemes for
phenomena all warrant intense study as poten- animal improvement.
tial tools for animal breeders. As this information Among the more important potential roles
accumulates it has important bearing on quanti- of cytogeneticists is the capability, using
tative and population genetic theory, requiring marker chromosomes that can be specially
modification of models and methods of animal produced or recovered from among spon-
breeding. The activities of the molecular and taneously occurring ones, to make experimental
cellular geneticists will yield clones of genes and tests of some basic assumptions upon which
other products as well as a host of ideas, many theory of animal breeding rests. They are
of which will deserve full-scale testing first in capable of detecting, for instance, deviations
laboratory animals and the most propitious in from the expectation that c h r o m o s o m e s
livestock. Such experimental work is best segregate randomly at meiosis or that sperma-
designed, conducted, and analyzed by animal tozoa bearing different haploid genetic contents
scientists. are equally able to effect fertilization and
support early embryonic development.
Biology of Reproduction Cytogenetic methodology is being applied to
and Animal Breeding provide partial answers to other fundamental
With the emergence of new techniques for questions confronting animal breeders. They
modulating the reproductive functions of can help detect the mechanisms responsible for
animals, manipulating their genomes, and the reduced reproductive performance that
altering their genotypes, modified strategies and frequently accompanies intense selection for
structures for animal breeding will need to be other production traits (31). They can contri-
explored. The explorations will first be theore- bute to an understanding of the decline in
tical; some will warrant further work involving fitness of inbred lines, particularly if such
complex simulations with computers. Ulti- decline is a consequence of mutants affecting
mately, new structures of animal breeding will the normal course of mitosis and meiosis (2).
need to be designed and institutions built to Ultimately cytogenetic techniques can provide
accommodate them. The traditional animal new sources of genetic variation for subsequent
breeding structure gave way to an entirely new exploitation by animal breeders. Already
one with the development of artificial insemina- triploid (1) and trisomic (5) chickens are
tion. When frozen semen became generally available for study. Tetraploidy-especially
available, the structure of the artificial insemin- aIlotetraploidy, which offers promise of provid-
ation industry was greatly changed. Even this ing entirely new forms of useful animals-is
industry has not been found to be the appro- worthy of intense investigation. So, too, is the
priate one, unless vast changes can be made further possibility of producing animals bearing
rather rapidly, for the widespread rational use a number of duplications, deletions, or other-
of embryo transfer. Just as animal breeders wise altered genomes.

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In conjuction with molecular and cellular ments in their own and related fields to de-
geneticists, the capabilities of the cytogeneticist partmental colleagues so that the outlook and
are required to assist with the mammoth task of understanding of newer developments of the
locating and mapping important genes and gene entire department is expanded. A portion of
clusters in livestock. Already a number of their time should be diverted from their own
laboratories are engaged in such work using basic research to collaborative efforts with
hybrid cell lines, in situ hybridation, and other animal scientists. Perhaps this activity would be
techniques. This work will require the diligent most useful it if were devoted to helping with
and sustained attention of a number of labora- the training of graduate students, who are
tories. under primary supervision of more traditionally
trained scientists.
A SUGGESTED AGENDA FOR What is visualized is that through constant
DEPARTMENTS OF ANIMAL SCIENCE
contacts and interactions, molecular and
Recent and continuing developments in cellular scientists would become familiar with
molecular, cellular, and developmental biology the larger concerns of the animal breeding and
vastly increase the potential for breeding production enterprise, and, equally important,
domestic animals for greater productivity. It is that animal scientists would become increas-
too early to know how the new knowledge will ingly cognizant of the techniques and the
be applied and in what order various potential knowledge of molecular and cellular biology so
new technologies will be exploited. But it is not that they could integrate them into their own
too early for all of those engaged in animal plans and activities and have at least a general
breeding and genetics to commence preparing appreciation of the power of adapting appro-
for fundamental changes in the nature of the priate aspects of the new biology.
enterprise. The most important step to be taken During an interim period, animal breeders
by departments of animal science that wish to and laboratory scientists will learn of each
be engaged at the forefront of animal breeding other's domains and build a mutual under-
research is to increase their capability to do standing of each others capabilities, limitations,
fundamental work in molecular and cellular and needs. Animal breeders will need to con-
genetics, reproductive biology, and other areas tinue to see to the continued growth of the
exhibiting promise of useful development. No industry, to monitor its progress, and to devise
single department can afford to develop pro- tools for its improvement. But they should give
ductive laboratories in all of such fields, but increasing attention to growth of the pool of
none can afford not to put a relatively high knowledge and technology being produced by
proportion of their resources into establishment the laboratory scientists so that they can plan
of one or more of them. for integration of appropriate segments in a
Faculty members in charge of the new timely and rational manner. They should
laboratories, whether they be new young also, from their vantage point, provide a greater
scientists trained outside of agriculture or perspective of the industry and explore and
established animal scientists with knowledge think about what developments would benefit
and skills to initiate a new research program, the industry most. There are vast amounts of
should be given special encouragement and work to be done and there will be too few
direction. They should be expected to initiate scientists to make advances on all fronts. The
and maintain a program of fundamental re- choice of specific objectives could be usefully
search that contributes important knowledge infuenced, therefore, by suggestions from
about genetic endowment of animals or its animal breeders.
organization, control, or functioning. It would Departments of animal science cannot
be well if a number of such laboratories con- expect to be given vast new resources with
centrated upon functions of production or which to embark on new ventures. For the
reproduction, but it is counterproductive to most part they will be required to realign a
exert undue influence on the direction of work rather static number of faculty positions to do
of individual scientists i n the academic com- so. To develop and exploit the new method-
munity. It is essential, however, that they have ologies it will be necessary to curtail or eliminate
the inclination and ability to convey develop- a number of traditional attitudes and activities

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and concentrate on doing those things that 10 Fechheimer, N. S. 1980. Cytogenetic developments
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