SCIENCE OF MEDICINE

 ƌŝĞĨ ,ŝƐƚŽƌLJ
ŽĨ ŶŝŵĂů MŽĚĞůŝŶŐ
by Aaron C. Ericsson, DVM, PhD, Marcus J. Crim, DVM
& Craig L. Franklin, DVM, PhD

ŶŝŵĂů ŵŽĚĞůƐ ƉůĂLJ Ă ďƐƚƌĂĐƚ first recorded instances of comparative

ĐƌŝƟĐĂů ƌŽůĞ ŝŶ ƚƌĂŶƐůĂƟŽŶĂů Comparative medicine is science were very observational, their
ƌĞƐĞĂƌĐŚ ĂŶĚ ĂĚǀĂŶĐĞŵĞŶƚ founded on the concept that purpose being to better understand
ŽĨ ŚƵŵĂŶ ĂŶĚ ĂŶŝŵĂů other animal species share human ontogeny and physiology.
ŚĞĂůƚŚ͘ physiological, behavioral, or Fortunately, many of the findings of
other characteristics with prominent thinkers like Aristotle
humans. Over 2,400 years ago it were documented and conveyed
was recognized that by studying to other countries via trade routes,
animals, we could learn much and animal modeling soon became a
about ourselves. This technique research tool of both European and
has now developed to the Arab physicians. While this early
point that animal models are period saw great discoveries, there
employed in virtually all fields of were still many misconceptions about
biomedical research including, the workings of the body, and it was
but not limited to, basic biology, not until the Renaissance (fourteenth
immunology and infectious through seventeenth centuries) that
disease, oncology, and behavior. animal modeling contributed to a true
paradigm shift in our understanding of
“Ought we, for instance (to give an human physiology.
illustration of what I mean), to begin by During the mid-sixteenth
discussing each separate species-man, lion, century, a few astute physicians such
ox, and the like-taking each kind in hand as Servetus and Lusitano deduced that
independently of the rest, or ought we rather blood followed two connected but
to deal first with the attributes which they distinct circuits through the body, i.e.
have in common in virtue of some common pulmonary and systemic circulation.
element of their nature, and proceed from In the late sixteenth and early
this as a basis for the consideration of them seventeenth centuries, William Harvey
separately?” (1578-1657) assiduously studied and
compared the anatomic and functional
-Aristotle (384 -322 BC) properties of the heart and vasculature
“On the Parts of Animals”
in multiple species including eels and
other fish, chicks, and pigeons. Based
Aaron C. Ericsson, DVM, PhD, Marcus J. Crim, EĂƌůLJ ,ŝƐƚŽƌLJ on these investigations, he penned
DVM, and Craig L. Franklin, DVM, PhD, are
in the Mutant Mouse Regional Resource ŽĨ ŶŝŵĂů MŽĚĞůŝŶŐ several seminal texts including De Motu
Center, ComparaƟve Medicine Program and The use of animals as models of Cordis in which he describes with great
Department of Veterinary Pathobiology,
University of Missouri.
human anatomy and physiology began accuracy, and in great detail, the human
Contact: [email protected] in ancient Greece (see Table 1). These circulatory system. He also pioneered

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Table 1. Early Milestones in Animal Modeling

at this time were
outbred and as
Years Researcher(s) Milestone the use of animals
6th c. BCE Alcmaeon of Croton Determined that the brain is the seat of intelligence and sensory
integration based on studies using dogs
became more
th
4 c. BCE Aristotle Studied embryogenesis and ontogeny in chicks experimental,
3rd c. BCE Erasistratus Studied the cardiovascular system in live animals and deduced rather than
that the heart functions as a pump
2nd c. CE Galen of Pergamum Studied cardiovascular and neuroanatomy extensively using live
observational,
animals researchers soon
12th c. Avenzoar Practiced surgical techniques on animals before applying them to appreciated the
humans, e.g. tracheotomy
17th c. William Harvey Studied anatomy of several species of live animals and provided confounding
accurate and detailed descriptions of the function of the factor of genetic
cardiovascular and other systems variability in
their research.
Through the
the theory of epigenesis, i.e. that embryos originate and efforts of many forward-thinking individuals such
develop from a single cell, based on his observations of as William Castle, Clarence Little, Halsey Bagg,
embryonic chicks (recommended for developmental studies and Leonell Strong, this problem was addressed
by Aristotle in Book II of The Generation of Animals). Of via inbreeding of mice to the point that genetically
note, Harvey was careful in his selection of model species, identical mice became available for experimental
in order to exploit certain properties of the animal such as
use (see Table 2). This provided a steady source of
heart rate and poikilothermy (“cold-bloodedness”).
research subjects that bred to maturity very quickly
The careful selection of the most informative
and with limited variability from litter to litter and
species for an animal model is still very important, but
year to year. As more and more inbred strains of mice
it also presents a unique challenge for investigators.
and rats were developed, it was soon appreciated that
Scientists must consider not only financial feasibility
there were inherent differences between strains in
and previous experiments utilizing a given species, but
basic biological parameters, as well as susceptibility to
also the unusual biological characteristics of a species induced and spontaneously occurring diseases. Many
and the available palette of imaging and molecular of these were complementary strains bred in parallel
techniques available for that species. The choice of a providing susceptible and resistant strains that are
naturally occurring species model, sometimes called otherwise genetically similar, such as the non-obese
the comparative method, was perhaps most famously diabetic (NOD) and related strains.3 Thus, strain
and succinctly stated by the 1920 winner of the Nobel selection is one of the most important considerations
Prize in Physiology and Medicine, August Krogh, in in animal modeling, particularly in rodents.
1929, “For a large number of problems there will be some If natural models were not available or feasible,
animal of choice or a few such animals on which it can be [most] the ability to manipulate the genome of a model
conveniently studied.”1 One recent example is the use of species allowed for the creation of animals uniquely
the nine-banded armadillo in studies of leprosy due to susceptible or resistant to a certain model. So, as
2
the armadillo’s unique susceptibility to M. leprae. advances were made in the field of genetics, scientists
became increasingly adept at manipulating the as
ŶŝŵĂů MŽĚĞůƐ ŝŶ MŽĚĞƌŶ ŝŽŵĞĚŝĐĂů ZĞƐĞĂƌĐŚ yet unsequenced genome of mice. The 1980s saw
By the beginning of the twentieth century, the use an explosion in this technology with the advent of
of animal modeling had increased dramatically and, transgenic mice carrying additional genetic material,
while some individuals still questioned the ethics of and knockout mice in which genetic material is
their use, animal modeling, particularly in rodents, deleted. Recently, our ability to manipulate the
had become the de rigeur method of demonstrating mouse genome has become increasingly refined with
biological significance. However, all research animals developments such as tissue-specific methods of

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knocking out Table 2. Recent Milestones in Animal Modeling

genes such as the
Years Researcher(s) Milestone
Cre-Lox system,4 1902 William Castle Begins breeding mice for genetic studies
methods of 1909 Clarence Little Begins inbreeding mice to eliminate variation
1920s Frederick Banting Isolated canine insulin and effectively treated diabetic dogs
turning on or off ca. 1930 Little and MacDowell First fully inbred mouse (20 brother × sister matings) achieved
gene transcription 1940s John Cade Studied the use of lithium salts as an anticonvulsant in guinea pigs and
translated his findings to treatments of depression
in vivo using 1976 Rudolf Jaenisch et al. Developed first transgenic mouse
tetracycline- or 1980s Several Extensive testing of drug safety and dosing regimens for HIV performed in
rhesus macaques
tamoxifen- 1987 Capecchi, Evans, and Developed first knockout mouse
induced systems,5 Smithies
1997 Wilmut and Campbell First animal cloned from an adult somatic cell, Dolly the sheep
and methods of 2002 Several Mouse genome sequenced
2004 Several Rat genome sequenced
identifying or 2009 Aron Geurts et al. Developed first knockout rat
removing entire
cell lineages in vivo
via fluorescent protein- and diphtheria-toxin receptor- ordered vertebrate possible to accomplish a given
knockin mice respectively.6, 7 Additionally, researchers scientific objective.
have used similar technologies to generate transgenic Additionally, the recognition of the impact of
8 9 10 11
rats, cats, dogs, rabbits, pigs, sheep, goats, cattle, the gastrointestinal and dermal microbiota led to the
chickens,12 zebrafish,13 and non-human primates,14 to birth of an entirely new research era – gnotobiotics.
name just a few. While the ability to generate targeted Through the use of Caesarian birth, flexible-film
gene knockouts in other species has lagged behind, isolator cages, and irradiated food, mice can now
knockout rats were successfully created in 2009 using a be maintained in completely germ-free conditions
zinc finger nuclease-based technique distinct from that or colonized with one or more defined bacterial
used in mice.15 species. A combination of eight commensal aerobic
The mouse continues to be the powerhouse and anaerobic bacteria called Altered Schaedler’s
for biomedical research (see sidebar page 206). Flora (ASF) is commonly used as the known intestinal
Undoubtedly, the most important change over the last microbiota.16 However, with the recent development
25 years is the spectacular escalation of the laboratory of robust methods of fingerprinting the entire gut
mouse in research, which stands in glaring contrast microbial community such as Denaturing Gradient
to the declining role of most non-rodent mammalian Gel Electrophoresis, Automated Ribosomal Intergenic
models (see Figure 1). By comparison, use of the Spacer Analysis, and deep sequencing, researchers
rat has plateaued, as targeted genetic manipulations are capable of quickly and reliably monitoring
proved more difficult in this species. The creation the composition of the gut microbiota and thus
of the first knockout rats may help to explain the moving away from more reductionist models such
very recent up-tick in rat model-based biomedical as ASF. While the development of inbred rodent
publications. However, with the rising capacity to strains allowed for the control of host genetics, the
modify the genomes of laboratory species other than development of research animals harboring complex
the mouse, the face of biomedical research is now but defined microbiota allows for control of microbial
changing. Genetically malleable species such as swine genetics known to impact host physiology. Moreover,
and the zebrafish are increasingly out-competing once gnotobiotics can also be applied to non-murine
common model organisms like the guinea pig, rabbit, species, so this field is likely to continue to evolve.
and ferret (see Figure 1). These important trends
reveal both 1) the dramatically increasing utility of FƵƚƵƌĞ ŽĨ ŶŝŵĂů MŽĚĞůŝŶŐ
certain model species relative to others, and 2) the What does the future hold for animal models?
refinement of animal research via use of the lowest As biomedical research funding agencies continue to

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with this disease, but

fail to develop the
devastating pulmonary
complications. To
circumvent these
deficiencies, a swine
model was recently
generated and early
data suggest that the
latter better replicates
pulmonary disease.17
Other examples include
the study of naturally
occurring diseases
in domestic species
that optimally mimic
disease such as the
study of osteosarcoma
progression and response
to therapy in dogs.18
This concept, referred
to as One Medicine,
promotes the sharing of
resources, knowledge,
and effort toward
Figure 1 the common goal of
Pubmed search results by publicaƟon date, 1970 through 2011. Search terms for each species included the improving the health
scienƟĮc name and the common name for each species͖ edžcept that only the scienƟĮc name was used for
mouse and rat. “Non-rodent mammalian models” includes the dog, rabbit, cat, rhesus macaque, guinea pig, and well-being of all
swine, chimpanzee, and ferret.
species and is proving to
be a powerful adjunct to
traditional laboratory animal models.
Humanized models such as transgenic
emphasize rapid and robust translatability of studies,
animals expressing human genes are also rising
it is likely that animal modeling will move more
to the forefront. A classic example involves the
and more towards models that most appropriately
mimic human conditions, using multiple models insertion of the gene encoding the human major
to ensure robustness of data and new genetic histocompatibility locus, HLA-B27 into rats.19
and metagenomic tools to develop and refine Individuals with this MHC haplotype have increased
“humanized models.” With advancements in susceptibility to several autoimmune conditions.
genetic engineering in non-mouse species, we are Similarly, rats with this transgene are more
also likely to see new models generated for diseases susceptible to autoimmune disease and as a result,
where mouse models have not adequately replicated this model has proven indispensable to studies of
the human condition. For example, genetically MHC-related disease susceptibilities. This concept
engineered mouse models of cystic fibrosis develop was expanded by coupling targeted mutations in
intestinal diseases similar to those seen in humans endogenous murine genes with the introduction of

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transgenes of mutated human genes. Newer models ZĞĨĞƌĞŶĐĞƐ

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Thus it is likely that animal models will continue
play a critical role in translational research and DŝƐĐůŽƐƵƌĞ
advancement of human and animal health. None reported. MM

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