1.

Animal models used in biomedical research: a theoretical approaches

Currently, laboratory animals (mice or rats) are used in basic as well as applied
research. Animal research includes the study of animal models to increase physiological
knowledge and identify new therapeutic targets or procedures. Furthermore, animal models
are used in the production and evaluation of therapeutic and diagnostic agents (such as
production of monoclonal antibodies, efficacy assessments of vaccines), and education (such
as training of different surgical procedures) (De Deyn and Van Dam, 2011).

The rationale for the use of animal models in biomedical research is that these
models allow the experimental study of the condition to a degree often impossible to human
subjects. Arguments regarding whether biomedical science can advance without the use of
animals are frequently mooted and makes as much sense as questioning if clinical trials are
necessary before new medical therapies are allowed to be widely used in the general
population (Chow et al., 2007). Animal models will be used in biomedical research until
science develops alternative models.

1.1. What is a model?

Before describing the notion of animal model we should emphasize the notion of a
model. Many authors have different opinions about the notion of model.

In Oxford English Dictionary is mentioned that a model is a representation of a real

or actual object (Chow et al., 2007). It means that a model not fully reproduce the
investigated subject and is not expected to be identical to the subject under investigation.

In a broad sense, according to Kaplan (1973) a model is described as follow: .we

may say that any system A is a model for the system B if the study of A is useful for the
understanding of B without regard to any direct or indirect causal connection between A and
B. In a more strict sense, according to Kaplan (1973) models are isomorphs of one
another. Both systems have the same structure, in the sense that whenever a relation holds
between two elements of one system a corresponding relation holds between the
corresponding elements of the other system. The systems need not stand in any causal
connection, for what is required is only that the relations correspond, and to satisfy this
requirement it is enough that we can put them into correspondence, that is, think of them as
corresponding.

Biomedical research models could also be analogues or homologous, respectively

(Chow et al., 2007).

Analogous modelsrelate one structure or process to another and are not unique to
biomedical research. Such models are also common in physics, engineering and
mathematics. A scaled-downmodel of an airplane is not an airplane but allows appreciation
of how the various parts ofthe structure relate to one another and how improvements may
be usefully made. Similarly,large animal models like the pig allow the development of new
minimally invasive surgicaltechniques and instruments.

Homologous models reflect counterpart genetic sequences and are only used in
biomedical research. Many animal models are both analogues and true homologues.

1.2. What is an animal model?

In biomedical research an animal is defined as a living organism with an inherited,

naturally acquired or induced pathological process that in one way or another closely
resembles the same phenomenon in man (Chow et al., 2007).

Consequently, in behavioral neuroscience, animal models are defined as living

experimental systems (Tamura et al., 1981), used to analyze brain-behavior relationships
under controlled conditions (van der Staay, 2000).

McKinney (1984) defined animal models as experimental preparations developed in

one species for the purpose of studying phenomena occurring in another species.

Both invertebrate and vertebrate animals are used as models in biomedical research.

Invertebrate models are useful in the fields of neurobiology, genetics and

development. Among them, notable example are Caenorhabditis elegans (Rudel et al., 2013)
and Drosophila(Chartier and Simonelig, 2013).
 Vertebrate models are responsible for many advances in biology and medicine and
are extremely important in translational research. This includes the use of both small animal
models (e.g. mice, rats, rabbits) and large animal models (e.g. dogs, pigs, monkeys) (Basu
and Wiklund, 2011).

There are many vertebrate models used for biomedical research as follow:

1. Pharmaceutical research including the development of biologics

2. Toxicology testing
3. Development and testing of new medical devices
4. Surgical research
a. the development of new surgical techniques such as techniques of
gastrectomy, open heart surgery, coronary artery surgery, microsurgery,
endoscopy and the use of arterial ligation in treating aneurysms;
b. the development of new therapies such as organ and tissue transplantations,
cardiopulmonary resuscitation.
5. Pathophysiological research
The quality or utility of an animal model depends on its validity. Moreover, a valid
animal model should present symptomatology, etiology and background (e.g. comparable
neuropathological, neurophysiological or electrophysiological features) and should display
concordant effects of therapy as the condition it is supposed to intimate.

The validity is necessary in homologous models. In these models, the symptoms as

well as the cause of the condition in the animal are identical to those of the human
condition. In behavioral neuroscience, the homologous models are very rare, and mostly
limited to well-defined lesion-syndromes.

In isomorphic models, although they display similar symptoms, the condition is not
provoked by the same events as the human condition, being limited to particular aspects.

Sometimes, researchers may seek to use animal models that specifically mimic
conditions of interests as opposite to using or developing general models. Such animal
models may either spontaneously mimic these conditions or be induced to simulate those
conditions (Chow et al., 2007).
 Spontaneous animal models arise through spontaneous mutation to mimic specific
conditions. Spontaneous genetic variants have been described in mouse and rat strains such
as Gunn rat (for hereditary hyperbilirubinemia) and the BB Wistar rats (for type I diabetes).

Induced animal models display the conditions as result of artificial manipulation,

such as surgical, chemical or genetic manipulations. In these models are included also
negative models.

The surgically induced model is a classical biomedical research model and was used
to understand brain function (rats, mice, nonhuman primates), develop organ
transplantation (dogs and pigs), and discover the role of insulin in diabetes (dogs) and to
develop cardio-pulmonary resuscitation (dogs).

The chemically induced models include the chemical ablation of the dopaminergic
neurons to create Parkinsonisms conditions (such as intranigral injection of 6-
hydroxydopamine, injections of lipopolysaccharide) (rats) and the use of carbon
tetrachloride to create cirrhosis (rats).

Transgenic animal models are important induced animal models. A transgenic animal
is one that carries a foreign gene that has been deliberately inserted into its genome (such as
B6;129-Psen1tm1Mpm Tg(APPSwe,tauP301L)1Lfa/Mmjax, a mouse model containing mutations
on the tau, presenilin 1 and APP gene simultaneously resulting in both tau and A pathology
in the brains).

In behavioral neuroscience, it have been develop animals models for neurological

and psychiatric disorders for a variety of conditions like peripheral neuropathy, myasthenia,
multiple sclerosis, stroke, epilepsy, amyotrophic lateral sclerosis, mental retardation,
dementia, Alzheimers disease, Parkinsons disease, AIDS-related dementia complex, pain,
head injury, schizophrenia, affective disorders, obsessive compulsive disorders, anxiety
disorders and stress.

The neuroscience community is far from reaching consensus about the level of
similarity between the brains and minds of humans and other species.

According to van der Staay (2000), animal models of behavioral dysfunction serve
two main aims:
 1. first, to enhance our understanding of the underlying substrates and mechanisms,
i.e. the brain-behavior relation. This is done experimentally by, for example,
inducing dissociations between processes, subprocesses and modulating
influences, either pharmacologically or through the destruction of neural tissue
(DMello and Steckler, 1996).
2. second, to assess the effects of putative neuroprotective, antidegenerative,
revalidation supporting, and/or cognition-enhancing compounds or treatments
(Allain et al., 1997).
The use of animal models had a huge impact in medical progress. Currently, there are
several international organizations that provide information about the use of animals in
medical research as follow:

Understanding Animal Research (UK)

Americans for Medical Progress (USA)
National Association for Biomedical Research (USA)
European Coalition for Biomedical Research (EU)
Searching for internet resources I observed that the website AnimalResearch.info
(2013) provide a lot of information about the reasons for using animals in research.

According to AnimalResearch.info (2013) there are four main reasons why animals
are used in research:

1. To advance scientific understanding

Adding to scientific knowledge through basic biological research helps us understand
how living things work, and apply that understanding for the benefit of both humans and
animals. The study of animals is a vital part of this research process. Many basic cell
processes are the same in all animals, and the bodies of animals are like humans in the way
that they perform many vital functions such as breathing, digestion, movement, sight,
hearing and reproduction. To treat disease, doctors and scientists must understand how the
healthy body works. This, in turn, leads to an understanding of what happens to the body
when we fall ill and how this can be put right.

A great deal of the knowledge of the body's anatomy and functions can be traced to
scientific findings from animal research. Comparing different species and studying the
differences and similarities between them is one way to gain insights. Even simple animals
can be used to study complex biological systems such as the nervous or immune systems,
which follow the same basic organization and function in all animals. For example, much has
been learnt about the function of neurons from studying the giant squid axon. Information
from this sort of work can then be applied to higher animals and humans.

2. As models to study disease

Humans and animals share hundreds of illnesses, and consequently animals can act
as models for the study of human illness. For example, rabbits suffer from atherosclerosis
(hardening of the arteries), as well as diseases such as emphysema, and birth defects such as
spina bifida. Dogs suffer from cancer, diabetes, cataracts, ulcers and bleeding disorders such
as haemophilia, which make them natural candidates for research into these disorders. Cats
suffer from some of the same visual impairments as humans. From such models we learn
how disease affects the body, how the immune system responds, who will be affected, and
more.

While contributing to our understanding of diseases, animal models also enable

researchers to explore potential therapies in ways which would be impossible in humans.
Studying disease mechanisms in animal models leads directly to the development of new
technologies and medicines that benefit both humans and animals.

Animals which are altered to create models of disease are known as induced models.
For example, surgery which damages a particular section of the spinal cord in rats gives rise
to symptoms like those seen in human patients with similar spinal cord damage. These
animal models help researchers understand what happens in the body following this type of
damage, and have been used in the development of new therapies.

Recent advances in genetic technology have allowed the development of transgenic

animals, which have new genes inserted into their DNA, allowing them to develop human
diseases which do not naturally affect them. In particular this has allowed mice to model
many human diseases which were previously difficult to study.

3. To develop and test potential forms of treatment

 Once researchers learn more about a particular disease, animals are used to develop
and test these potential therapies as part of the applied research process. For example,
medicines for Parkinson's disease have been developed using animal models with induced
Parkinson's-like symptoms. Models such as these are an essential part of applying biological
research to real medical problems, allowing new targets for disease intervention to be
identified. Data from animal studies is essential before new therapeutic techniques and
surgical procedures can be tested on human patients.

Diagnostic tools such as scanners, and implants such as heart pacemakers or artificial
hips, are safe and effective only because they were developed and tested in animals. Many
surgical techniques, such as open heart surgery and heart transplants, rely on methods and
equipment that were developed using animals.

4. To protect the safety of people, animals and the environment

New medicines require testing because researchers must measure both the
beneficial and the harmful effects of a compound on a whole organism. A medicine is initially
tested in vitro using tissues and isolated organs, but legally and ethically it must also be
tested in a suitable animal model before clinical trials in humans can take place.

The animal tests provide data on efficacy and safety. They not only identify potential
safety concerns, but also determine the doses which will be given to volunteers and patients
during the first human trials.

Testing on animals also serves to protect consumers, workers and the environment
from the harmful effects of chemicals. All chemicals for commercial or personal use must be
tested so that their effect on the people and animals exposed to them is understood. The
chemicals that we use day-to-day can accumulate in the water, ground or air around us, and
their potential impact on the environment must be researched thoroughly.

Consequently, AnimalResearch.info (2013) provide a list with those disease which are
now prevented or cured with the knowledge obtained by animal experimentation. The
importance of animal models in the medical progress is further exemplified by the list of
Nobel prizes awarded for Physiology and Medicine (89 Nobel prizes were directly dependent
on animal-based research).
 Finally, all animal models used in biomedical research have their limitations. These
limitations are, however, an intrinsic part of all modelling approaches that use surrogates
and do not render the scientific method invalid.

My scientific achievements are mainly contributions in the field of neurophysiology of

cognitive processes, especially learning and memory processes, using pharmacological and
toxicological animal models of human neurological disorders, with relevance for biomedical
research.

The main focus in this thesis is on my contribution to explain behavioral and

molecular mechanisms of experimental animals behavior to gain new insights for
understanding the mechanisms of human neurological diseases.

In the next parts of the present thesis, I will present, starting with 2007,my personal
contributions on elucidating of behavioral and molecular mechanisms of experimental
animals behavior achieved following the PhD award in 2006.