MENTAL RETARDATION AND DEVELOPMENTAL DISABILITIES
RESEARCH REVIEWS 2:249-256 (7996)
ANIMAL MODELOF AUTISM
BASED
O N DEVELOPMENTAL
DATA
Patricia M. Rodier"
Department of Obstetrics and Gynecology, University of Rochester, Rochester, New York
Animal models of human brain damage syndromes range
from ones that are purely theoretical t o ones that attempt t o
duplicate a human syndrome a t every level. The similarity between
animal and human cases in experimental manipulation, CNS alter-
ation, and tests of parallelism is sometimes restricted by the extent
t o which experimental species match humans in their basic biology.
However, the information available from the human condition is
often the most important limiting factor in the degree t o which
similarities can be achieved. The recent discovery o f a high rate o f
autism in patients exposed t o thalidomide around the time o f neural
tube closure suggests an appropriate experimental manipulation for
an animal model of the disease. Rats exposed t o valproic acid a t the
same stage of development mimic the thalidomide cases' apparent
injuries t o the brain stem. Whether the behavior o f these brain-
damaged animals is similar t o that observed in autism is difficult
t o determine, because the diagnostic behavioral features of the
disease are not easily translated into animal tasks. However, ana-
tomical observations in human cases provide tests of parallelism
that can be applied t o animals, and these appear t o be positive in
the animal model. The development o f behavioral tests that dis-
criminate "autistic behavior" from that o f other brain damage
syndromes may require further research in both humans and ani-
mals. o 1997 Wiley-Liss, Inc.
MRDD Research Reviews 2:249-256, 1996.
Key Words: autism; teratology; valproic acid; thalidomide; cranial
nerves; brain stem
o qualify as a model of a human disease or syndrome, an
T animal model must exhibit characteristics that resenible
those of the condition under study in some way, but the
points at which the similarity is confirmed vary from one model
to another. The purpose of this review is to demonstrate some of
the variety among whole animal models that have been used in
the study of mental retardation and related conditions and to
describe a new inodel developed for investigation of autism.
Comparison of existing models should serve to remind
us that animal models of brain damage syndromes can be
constructed in many different ways. They may start with the
behaviors of interest and be used to investigate the underly-
ing brain abnormality. They may start with the creation of a
suspected brain injury and be used to test what behavioral
anomalies result. They may start with a method of injury, such as
exposure to a suspected teratogen, or the targeted knockout of a
particular gene, and be used to investigate effects on the brain
and functional consequences.
The four models to be compared were selected because
they demonstrate differences in approach to modeling brain
injuries. Three have been used to study specific human
o 1997 Wiley-Liss, Inc.
syndromes-fetal alcohol syndrome (FAS), Lesch-Nyhan syn-
drome, and phenylketonurea (PKU)-and one is a more general
model of developmental brain injury, exposure to the antinii-
totic agent, methylazoxyniethanol (MAM). FAS is characterized
by minor malformations of the face along with mental
retardation. Victims are typically aEable and better at verbal than
quantitative tasks. Researchers have only begun to unravel the
underlying CNS injury and the exposure conditions that are
required to produce the syndrome. Lesch-Nyhan syndrome is a
congenital disease characterized by self injury, especially biting of
the hands and lips, aggressive behavior, and varying degrees of
mental retardation. The genetic defect that produces the
syndrome is known, but the pathways by which the genetic
anomaly produces the symptoms are unclear. Development of
treatments depends on determining the sources of the syniptom-
atology. The mental retardation associated with PKU has long
been known to arise from various inborn defects of liver
nietabolism that lead to elevation of phenylalanine levels and
depression of tyrosine levels. Untreated, the victims suffer severe
brain damage and severe mental retardation. In this case an
effective treatment has already been established-a diet low in
phenylalanine allows patients to develop relatively normally.
The major question remaining for human and animal studies is
whether dietary restriction can eliminate completely the
brain-damaging effects of the metabolic disorder, and what level
of restriction is necessary to achieve the optimal result. MAM is
an antimitotic agent used to eliminate neurons as they are
forming. Brief exposures during brain development can model
the effects of a host of conditions that block cell proliferation.
Because the effects of blocking cell production depend on the
stage of development when exposure occurs, many different
patterns of brain damage can be induced with MAM.
COMPARISON OF MODELS
Experimental Manipulation
Figure 1 represents some features of animal models in a
general way and Figure 2 represents those features for the specific
models reviewed in this article The box a t the top stands for the
experimental manipulation of the animals, which might involve
treatments as diverse as a chemical exposure, the selection of a
*Correspmdeiicr: to: Patricia M . Rodicr, Ph.13.. Department of OB/C;YN, Box 668,
Medical Center, Univeruty of Rochester, Rochrstcr, NY I-lh.12. E-m.iil:
rodicrp(~ehsct7..enviiied.rochestcr.rdu.

;enera1 Features of Animal Models
experimental
manipulation
1 alteration of CNS
I
structural, chemical, or
functional expression
of injury
investigation
of unknowns
Figure 1. One way t o conceptualize an
animal model of a human behavioral
abnormality, such as mental retarda-
tion, is t o consider how it is created,
how the experimental manipulation
changes the CNS, what symptoms can be
tested for parallelism, and what un-
knowns are to be investigated.
strain with special genetic characteristics,
or a surgical lesion. While this step is
sometimes designed to reproduce the
events thought to cause the condition
being modeled, it is important to note
that this is not always so. Among the
models outlined in Figure 2, models of
FAS involve exposure to the same
teratogen identified as injurious in human
cases, but developing humans are never
exposed to 6-hydroxydopamine, alpha-
methylphenylalanine, or MAM. These
experimental manipulations have not
been used because they mimic human
exposures, but because they create bio-
logcal abnormalities associated with
changes in the central nervous system
known or suspected to play a role in
behavioral symptoms.
The 6-hydroxydopamine model of
Lesch-Nyhan syndrome and the alpha-
methylphenylalanine model of PKU are
ones that required extensive knowledge
of the diseases before they could be
created. In contrast, teratologic exposure
models may appear on the surface to be
easy to establish. In fact, even the attempt
to model known teratogens is a complex
process. W e cannot claim to reproduce
human exposure conditions unless we
know the details of human exposure
patterns, details of the human pharmaco-
kinetics of the agent, and the tissue
concentrations of the proximal teratogen
250
at the target organ. Without these data,
we cannot determine whether our animal
exposures are comparable to human
cases. In the case of ethanol, years of study
indicate that many species exhibit at least
parts of the syndrome observed in hu-
mans [Driscoll et al., 19901, but none of
our models does more than approximate
the pattcrns of human consumption,
because experimental animals will not
willingly consume large quantities of
ethanol. Thus, accurate models of terato-
logic exposure are sometimes difficult to
produce. Just as with models of human
diseases, teratologic models require exten-
sive preliminary study in humans and
animals to assure comparability between
animal and human results.
The classic example of a compound
for which the assumption of comparabil-
ity was erroneous is thalidomide. It was
approved as safe on the basis of rodent
tests and discovered to be a potent
teratogen in primates only after thousands
of human cases of birth defects were
traced to the drug [Lenz, 19621. A-
though the pharmacology of thalidomide
is not known, it is suspected that
susceptible species produce an unknown
metabolite that is the proximal teratogen,
while resistant species metabolize thalido-
mide to less toxic products.
Models of Lesch-Nyhan syndrome
provide another example of the fact that
recreating the conditions thought to
cause a syndrome is not a guarantee of an
optimal model. The human syndrome is
known to be caused by a genetic
deficiency of hypoxanthine phosphoribo-
syl transferase (HPRT) [Seegmiller et al.,
19671, yet transgenic mice deficient in
H P R T do not seem to reproduce either
the severe dopamine deficiency or the
behavioral abnormalities observed in the
human condition Uinnah et al., 1994,
19911, while rats treated neonatally with
6-hydroxydopaniine exhibit both [Breese
et al., 19841. The difference between the
dopamine effects of the two models
appears to be a matter of degree, with
6-hydroxydopanine treatment reducing
levels to approximately those observed in
human cases [Lloyd et al., 19811, and the
genetic model creating a less severe
deficiency. Thus, for studying some
aspects of the Lesch-Nyhan syndrome,
the 6-hydroxydopamine model is more
valuable than the model closer to the
cause of the disease.
Both mutant mice [Shedlosky et
al., 19931 and rodents treated from
infancy to inhibit metabolism of phenyl-
alanine to tyrosine [Greengard et al.,
19761 have been shown to model the
elevated phenylalanine and depressed
MRDD SEARCH REVIEWSA N I M AMODEL
L OF AUTISM RODIER
tyrosine levels of PKU, and the CNS
effects in the two types of models are
thought to be the same. Because the
genetic model exposes the subject to
PKU before and after birth, while the
pharmacological model creates PKU con-
ditions only after birth, it may be possible
to compare the two to discover whether
prenatal exposure causes damage that
cannot be alleviated by postnatal dietary
restriction. For many purposes, however,
these models seem so similar in their
effects that the choice between experi-
mental manipulations can be made on the
basis of cost, ease of adjusting transmitter
levels, etc., rather than which cause is
most similar to the human condition.
Methylazoxymethanol(MAM) mod-
els are good examples of ones that are
theoretical at every point except the
behavioral outcome. That is, in this case
the experimental manipulation is not the
same as any human insult, and the
resulting brain injury is not known a
priori to be similar to any human
disorder. Yet exposure to MAM can
produce many behavioral syndromes of
great interest to students of brain damage.
The investigator hypothesizes that injury
to some part of the nervous system causes
mental retardation, then uses MAM,
X-ray, or other antimitotic agents to
injure that region developmentally [re-
viewed in Rodier, 19861. MAM expo-
sure is not a cause of any human cases, but
it leads to a defined brain injury that can
be directed by the timing of exposure to
involve the region selected. The model
animals can then be tested for deficits on
tasks believed to be diagnostic of the
syndrome of interest. Because MAM
represents many kinds of teratogens that
injure neurons as they form, the treat-
ment models many teratogens, and is a
valuable tool for investigating which
brain regions are responsible for particular
behavioral symptoms. To the extent that
MAM-exposed animals exhibit behav-
ioral symptoms like those seen in mental
retardation, they can tell us what patterns
of neuron loss may underlie M R syn-
dromes. These models differ from the
others under discussion in that they are
not directed at only one type of brain
damage but can be adjusted to mimic
many different syndromes, including ones
not yet recognized, but theoretically
possible, in humans.
In summary, the experimental ma-
nipulation involved in a model may be
extremely similar to the one known to
cause the condition of interest, or it may
be quite different. Models using manipu-
lations different from that known to cause
the condition niay be preferred for many
 - Models of Fetal
Alcohol Syndrome

prenatal exposure
to EtOH in liquid diet
Model of Lesch-Nyhan
Syndrome

neonatal
6-hydroxydopamine
Models of PKU

neonatal methylphenyl-
alaninel HPH-5 mutant
r
MAM Models

cell loss and aberrant phenylalanine excess reduction of neuron

’
connections observedl
other effects on CNS
development not known
I DA neuron deficit
I
and tyrosine deficit in
blood (mechanism of
CNS injury not com-
numbers in regions
forming during
exposure
pletely known)

reduced contrast behavioral effects

abnormal suckling and
growth/ deficits in
response inhibition and
many behavioral tests
(SIB)/dopamine deple- sensitivity/ deficits on,
tion in cortex and stria- dependent on which
tasks requiring frontal regions were injured
turn/ receptor changes cortex, but not others

behavioral mechanisms environmental triggers structural-functional

mechanisms of EtOH
action/ endocrine
effects/ neuroanatomyf
of SIB/ transmitter and
receptor changed
therapy
mechanisms of injury/
Phe dose threshold I correlations/ behavior-
al mechanisms1 bio-
chemical effects

Figure 2. Four types of models that have been studied by many investigators are outlined according t o the scheme described in Figure 1.
Models of FAS, Lesch-Nyhan syndrome, and PKU were developed for study of specific human conditions. MAM models involve injuries to
the developing CNS targeted t o specific brain regions or systems. They allow investigators t o test many general hypotheses about
developmental brain damage.

reasons, including: to avoid some lack of
correspondence between animals and
humans, to increase the experimenter’s
control of dose, or to focus on some
known or suspected feature of a condi-
tion. In any case, the value of a model
cannot be judged by how closely the
manipulation matches the cause of hu-
man cases. Rather, it is the degree to
which the outcome of the manipulation
parallels human cases that is important.
CNS Alteration
The second box represents the
change in the nervous system that follows
the manipulation. This box is rarely filled
in completely in models of mental
retardation. Even in the chemical treat-
ment models in Figure 2, we are not
certain of all the details of how the CNS
is altered, even though much is known
about the chemical changes that lead to
damage and those that follow it. Since
our knowledge of the CNS injury in
most human conditions with mental
retardation IS limited or nonexistent,
there is rarely a chance to confirm in an
animal model some known human effect.
Without information, the neuroanatomy
MRDD RESEARCH MODEL
REVIEWS ANIMAL OF AUTISM RODIER
or neurochenlistry or physiology of the
CNS often become unknowns that we
hope will be revealed in the model. The
lack of human CNS data also encourages
the use of models in which the CNS
effects are inferred from the fact that
behavior is affected.
Tests of Parallelism
Box 3 includes examples of mea-
sures of expression of brain damage used
in various models. These are the tests that
have been used to confirm the correspon-
dence between the models and the
condition under investigation. Some are
very general. For example, in FAS
studies, a wide variety of behaviors have
been shown to be abnormal after prenatal
exposure of rodents to ethanol but few of
these measures have been investigated in
human cases. Conversely, many measures
of ethanol’s effects widely used to assess
human cases cannot be applied to animal
models. If we are willing to compare
more general categories ofbehavior (e.g.,
a maze task as a measure of learning versus
school performance as a measure of
learning) then the correspondence be-
tween humans and animals is reasonably
good. There is no question that animals
prenatally exposed to ethanol are brain-
damaged. What is harder to test is the
extent to which their injury duplicates
the one in human FAS cases, as opposed
to other kinds of brain damage. The
abnormalities of facial development char-
acteristic of FAS provide another test of
some rodent models [Webster et al.,
1983; Edwards and Dow-Edwards, 19911,
and support the idea that ethanol’s
teratologic effects are similar across spe-
cies.
The behavioral data for the PKU
models are unusually specific, in that
Diamond and her colleagues have worked
back and forth among species, using tests
that are similar in the brain regions on
which they depend [e.g., Diamond, 1994
and Strupp and Diamond, 19961. The
ability to monitor blood levels of the
abnormal amino acids of the condition,
the knowledge that these are the caus-
ative factors in the brain injury, and the
specificity of the behavioral tests make
this model virtually complete. That is,
there is no reason to think that anything
we discover in animals with excess
phenylalanine cannot be applied to the
251
human case. Most models are much more
limited. However, a model need not
match every aspect of the condition to be
useful; it need only allow us to learn new
things about the condition ofinterest. For
example, while the 6-hydroxydopamine
model of self-injurious behavior (SIB)
does not purport to reproduce all the
brain abnormalities of the genetic syn-
drome of Lesch-Nyhan, it does repro-
duce the dopamine depletion that appears
to be the abnormality underlying SIB.
Thus, while this model would have
limited value for studying the neuro-
anatomy of Lesch-Nyhan syndrome, it is
very useful for studying the environmen-
tal triggers of SIB [Tessel et al., 19951, or
testing drug therapies for SIB [Breese et
al., 19861.
It is impossible to overemphasize
the importance of human data in the
development of animal models. Our
ability to confirm an animal model is
totally dependent on the existence of
specific tests of the human condition that
discriminate it from others and that can
be applied to animals. For example, a
hypothetical human condition defined
only by low I Q cannot be modeled,
because we can never prove that an
animal exhibits low IQ. Even ifwe could,
so many conditions include low I Q that
we could never be certain that our aniinal
model was the same as the hypothetical
condition, rather than some other syn-
drome. O f course, it is possible to
imagine a condition that can be defined
only in human terms-e.g., stuttering-
and to imagine that all the details of the
initiating cause, CNS anomalies, associ-
ated problems, and therapies could be
worked out in humans without resorting
to the use of animal models. However,
when animal models are desired, then we
must encourage the collection of human
data that could provide tests of parallel-
ism.
Unknowns to be Investigated
Box 4 lists some typical unknowns
under study in the models. Just as the
models vary in the information on which
they are based, they vary in what
questions they can be used to address.
AN ANIMAL MODEL
OF AUTISM
Autism is a disease defined by
deficits in social interactions and conimu-
nication, as well as repetitive behaviors
and restricted interests, according to the
DSM-IV [American Psychiatric Associa-
tion, 19941. Although the syndrome
occurs with normal and even high
intellectual function, about 75% of cases
252
exhibit mental retardation [Rutter and
Lockyer, 19671. The diagnosis requires
that symptoms be present by the age of
three, but little is known about when the
disease begins or what its causes might be.
Estimates of the incidence of the disease
vary, but a prevalence study in Nova
Scotia indicates a rate of more than 1 per
1,000 [Bryson et al., 19881.
“It is impossible to
overemphasize the
importance of human
data in the development
of animal models. Our
ability to contrm an
animal model is totally
dependent on the existence
of spec@ tests of the
human condition that
discriminate it from
others and that can be
applied to animals. ”
The creation of an animal model
has been problematic for many reasons.
The diagnostic criteria are too general
and too focused on human behaviors
such as speech to be easily adapted for
animal testing. More specific functional
abnormalities have been reported to be
associated with autism. For example,
seizures are coninion [Olson et al., 19881,
but these are not diagnostic, since they
occur in some people who do not have
the disease and fail to occur in some who
do have the disease. Some experimental
lesions lead to behaviors that may be
related to autism. For example, neonatal
lesions of the limbic system in monkeys
produce a syndrome with lack of facial
expression and aberrant social behavior,
which could be related to similar symp-
toms in autism [Bachevalier and Merja-
nian, 19941. O n the other hand, the
alterations of recognition memory in this
model seem different from the deficits
reported in autism in that they interfere
with recognition through all sensory
modalities, while the defects reported in
autism appear to be restricted to auditory-
verbal material [reviewed in Killiany and
Moss, 1334].
Anatomical studies of autopsy cases
have not provided results that are easily
related to the symptoms of autism. No
REVIEWS ANIMALMODEL
MRDD RESEARCH
differences have been demonstrated in
studies of the cortex [Williams et al.,
1980; Coleman et al., 19851. One lab has
reported an increase in cell packing
density in the limbic system [Kemper and
Bauman, 19921 and several have observed
reductions of cell numbers in the cerebel-
lum [Bauman and Kemper, 1985; Ritvo
et al., 19861. While the number of cases
studied to date is very small, and further
research might reveal more subtle abnor-
malities, the present data rule out any
large or obvious lesions [reviewed in
Bauman and Kemper, 19941. Given the
absence of histological changes in most of
the brain, it is not surprising that MRI
studies in living subjects have often been
negative [Kleiman et al., 1992; Piven et
al., 19921. However, several groups have
reported reductions in the area of the
posterior lobe of the cerebellum in
midsagittal sections [reviewed in Cour-
chesne et al., 19941, and the largest study
confirms that finding and adds small but
significant reductions of the midbrain,
pons, and medulla [Hashinioto et al.,
19951.
A new discovery suggests that it
might be possible to model the kind of
developmental accident from which au-
tism arises. While studying ocular motil-
ity in victims of prenatal thalidomi.de
exposure, Miller and Stromland [ 19931
recognized that a surprising number of
the cases were autistic. They saw four
cases among the 86 patients from the
Swedish regstry they examined, and
found a fifth in records of the fourteen
unavailable for study [Stromland et al.,
19941. The external malformations caused
by thalidomide have been studied exten-
sively, making it possible to link particu-
lar abnormalities to injury at particular
stages of development [reviewed in Miller,
19911. (Most of the thousands of women
who took the drug during pregnancy
took it for weeks or months, but some
took it briefly, and a few only once. This
has made it possible for teratologsts to
piece together a timetable for defects
caused by thalidomide exposure.) When
the cases in the Swedish registry were
classified by time of injury, all the autistic
cases fell into the group exposed between
the 20th and 24th days of gestation. Only
15 cases had evidence of such early
exposure [Miller, 19911. Thus, the rate of
autism in the early exposure group was
one in three.
The results of the thalidomide
study provide the information needed to
create an animal model of autism. They
pinpoint the time of injury and id en ti^
one teratogen with the potential to cause
the disease. W e do not know what the
OF AUTISM RODIER
 histology of the brains of these living
cases is like, but each patient has neuro-
logical symptoms indicative of a brain
stem injury, expressed as malfunctions of
several motor cranial nerves. In addition,
each has hearing deficits and malformed
ears [Stromland et al., 19941. The neuro-
logical defects of the cases make sense in
the context of the stage of development
identified, because it is the stage when
motor neurons of the cranial nerve nuclei
form (Bayer et al., 19931. The results also
suggest that structures outside the brain
stem could not have been injured directly
by thalidomide, because only structures
of the midbrain, pons, and medulla form
so early. The 20-24 day period precedes
the initiation of neuron production for
the cerebellum or limbic system [Bayer et
al., 19931. Thus, the thalidomide results
suggest that the primary injury underly-
ing autism is centered in the early-
fomiing parts of the brain stem, with the
abnormalities of other structures repre-
senting secondary effects.
O n the surface, the idea that autism
involves an injury of the brain stem might
seem at odds with the current interest in
the cognitive impairments in the disease.
However, there is no conflict between
the thalidomide results and cognitive
effects. A developmental injury that
disrupts the brain stem might well alter
development of higher centers forming
subsequent to the injury. Further, dis-
turbed input from the brain stem could
lead to malfunctions a t any level of the
CNS, even if development after the
injury proceeded normally. The thalido-
mide results do not rule out malfunctions
of any area of the brain; they only rule out
a direct injury to any but the earliest-
forming neurons.
One thing that makes the new
thalidomide data seem surprising is that
neuroteratologists have often assumed
that the period around closure of the
neural tube is of interest only with regard
to neural tube defects. The idea has been
that such early injuries are more likely to
result in death or full recovery than in
brain-damaged survivors. There are prac-
tical reasons for studying later CNS
injuries, too. Early exposure to teratogens
runs the risk of injury to other organs,
confounding brain damage with somatic
problems. The result of these consider-
ations has been that we have studied fetal
and postnatal insults to the brain, and
avoided embryonic ones. In fact, the
cranial nerve effects of thalidomide were
noted as long ago as 1964 [d’Avignon and
Barr]. Thus, long before thalidomide was
linked to autism, the data indicated that
the earliest stages of brain development
MRDD RESEARCH
REVIEWSANIMAL
MODEL
OF AUTISM RODIER
were sensitive to this agent, and probably
many others.
Unfortunately, as mentioned ear-
lier, thalidomide’s effects in rodents are
different from its effects in primates
[Schumacher et al., 19681. Therefore, to
model the brain injury of the thalidomide
cases, we used valproic acid (VPA). This
anticonvulsant produces a sequence of
malformations similar to those of thalido-
mide and the effects are seen in all species
tested [Binkerd et al., 19881.VPA is trans-
ferred to the embryo along with its main
active metabolite, 2-en-VPA [e.g., Nau,
19861. The metabolite is not teratogenic
[Nau et al., 19841 while VPA is highly
teratogenic in embryo culture as well as
in vivo [Kao et al., 19811 suggesting that
VPA is the active teratogen. VPA expo-
sure has been linked to both neural tube
defects [e.g., Ehlers et al., 19921 and au-
tism [Christianson et al., 19941.
“The results of the
thalidomide study provide
the information
- needed to
create an animal model of
autism. They pinpoint
the time of injury and
identify one teratogen
with the potential to
cause the disease. ”
Experimental Manipulation
W e tested whether it is possible to
disrupt the formation of the motor cranial
nerve nuclei as the neurons are forming
and leave the brain-damaged animals
viable [Rodier et al., 19961. Counting the
day when a plug was found as the first day
of gestation, we targeted the stages of rat
gestation when neurons first appear
[Bayer et al., 19931 treating on day 11.5,
12, or 12.5 with 350 mg/kg of VPA via
IP injections to the dam. Control and
treated offspring were killed 10-11 days
after birth and their brains prepared for
study of the cranial nerve nuclei.
CNS Alteration
Counts of motor neurons showed
that the earliest treatment time affected
only the earliest-forming nuclei, the
motor nucleus of trigenllnal and the
hypoglossal nucleus. At the next treat-
ment time, neuron numbers in these
nuclei and the abducens nucleus were
reduced. Only at the latest time were the
neuron numbers of the oculoniotor
nucleus affected, along with the three
other nuclei affected at earlier treatment
times. None of the treatment times was
late enough to involve the facial nucleus,
which forms slightly later and over a
longer period than the other nuclei
studied. Reductions in the number of
neurons in motor nuclei persisted in rats
held to 60 days.
The great specificity of VPA effects
with sniall changes in exposure time is
typical of many teratogens with antimi-
totic action [reviewed in Rodier, 19861.
However, VPA’s action appears to be
selective on some other basis in addition
to time, because we found no effects on
neuron number when we counted three
cell groups that form during the exposure
period but contain no somatic motor
neurons. These were the locus ceruleus,
the dorsal motor nucleus of the vagus,
and the mesencephalic nucleus of trigemi-
nal.
How well do the VPA-exposed
rats reproduce the features of the thalido-
nllde cases? They have lesions of the
cranial nerve motor nuclei and they lack
limb malformations, like the human
cases. However, none of the animals had
ear malformations. Since ear malforma-
tions have been shown to occur in rats
exposed as embryos to higher doses of
VPA and sacrificed just before birth
[Binkerd et al., 19881, it may be that our
model could be improved by using a
higher dose or multiple doses, but so far
we have been unable to reproduce the ear
effects even at doses with low survival
rates.
Tests of Parallelism
The next step in developnient of a
model of autism is to show that the
animals resemble human cases in ways
other than those the treatment was
designed to produce. In particular, we
would like to be able to demonstrate that
their behavior is like that of people
diagnosed as autistic. T o do this, of
course, requires that we can determine
which human behaviors discriminate
between people with autism and others
and compare those behaviors in treated
rats and controls. As mentioned earlier,
this is not a simple task. For cxaniple, we
cannot look for delayed speech in an
animal model, but we could study
vocalizations in developing rats [Adams et
al., 19831. I f w e had data to indicate that
rat vocalizations are analogous to human
speech, that would make testing them
worthwhile. Alternatively, if we had data
indicating that human vocalizations other
than speech are disturbed in autism, we
253
could develop a test for animals. The
needed studies could be done, but they
have not been done.
An extremely concrete and highly
discriminating behavioral characteristic of
autistic children is their failure to point, as
described by Osterling and Dawson
[1994]. W e have not been able to think of
animal behaviors used to draw another
animal’s attention to something-the
essential feature of this kind of pointing.
Even “pointing” in dogs, while it com-
municates information to the master,
does not seem to be a behavior initiated
by the dog for purposes of communica-
tion; it may be more related to freezing
behavior.
Social behaviors, as a general cat-
egory, are affected by many kinds ofbrain
damage. For example, developmental
exposure to methylmercury [Burbacher
Model of Autism
VPA during neural
tube closure
reduction of neuron
numbers in cranial
nerve motor nuclei
I
changes in‘posterior
lobe of cerebellum,
shortening of brain
stem
primary and secondary
effects on anatomy/
connections/ behavior?
Figure 3. A new animal model of au-
tism is based on human evidence that
the disorder can be induced at a very
specific stage of CNS development. The
model has yet to be tested for behav-
ioral outcomes, but has been tested for
the similarity of its neuroanatomical
characteristics t o those reported in hu-
man cases of autism.
254
et al., 19901, developmental exposure to
lead [Laughlin et al., 19911, maternal
deprivation [Seay and Harlow, 19651,
and amygdaloid lesions [Kluver and Bucy,
19391 all disrupt social behavior. To use
abnormalities of social behavior in ani-
mals as confirmations of parallelism to
autism we would need to identify social
behaviors affected in autism, but not in
other brain damage syndromes.
Stereotypies occurring naturally in
some dogs respond to the same drugs
effective in reducing obsessive-compul-
sive disorder (OCD) in humans [Rapoport
et al., 19921. Recent drug treatment
studies of autistic subjects suggest that
O C D symptoms in autism are related to
the O C D symptoms in other human
syndromes [Gordon et al., 19921. If this is
the case, these behaviors-very common
in mice-might be useful in developing
animal models ofautism. Again, we need
more information. W e feel certain that it
will be possible eventually to identi@
behaviors that can serve to confirm an
animal model, but it may require new
studies of humans as well as new studies
of animals.
While it is frustrating to find no
ready test of autistic behavior for animals,
it is noteworthy that several of the models
diagrammed in Figure 2 have included
tests of parallelism that are not behavioral.
Until sound behavioral tests can be
identified, there is at least one set of
findings that provides specific test hypoth-
eses for comparing an animal model of
autism to the human condition, and that
is the abnornialities of cerebellar anatomy
reported in histology and imaging studies.
Because we know that no cerebellar
neurons form as early as the treatment
times used in the VPA model, and thus
cerebellar neurons could not have been
injured directly by exposure, any abnor-
malities observed in the cerebellum
would not fit into the “CNS alteration”
part of the model, as do the effects on
cranial nerve motor nuclei. Rather, such
results would be a confirmation of the
parallelism between the animal model
and the human condition. In fact, cell
counts of Purkinje cells and neurons of
the deep nuclei, and measures of the size
of the cerebellar lobules in young animals
and adults indicate that VPA exposure on
day 12.5 of gestation leads to changes in
the cerebellum that are like those ob-
served in human cases of autism [Ingram
et al., 19961. Not only are the Purkinje
cells reduced in number, but the effect in
the animal model is localized to the
neocerebellum. The size difference is also
present only in the posterior lobe, just as
in MRI studies. The deficits in cell
MRDD RESEARCH
REVIEWS ANIMAL
MODEL
OF AUTISM RODIER
numbers in the deep nuclei exist in the
nucleus interpositus, corresponding to
the globose and emboliform nuclei af-
fected in human cases, while the dentate
nucleus is spared in the animal model as it
is in humans [Bauman and Kemper,
19941. The fastigial nucleus is cell poor in
autistic brains, but not significantly af-
fected in the rodent model. With this
small exception, the pattern of cerebellar
effects observed in the model is amazingly
similar to that reported in autism. Since
the effects in the animal cerebella must be
secondary to the injury of cranial nerve
motor nuclei, we now suspect that the
cerebellar abnormalities of the human
cases are likely to be secondary effects of
an injury to earlier-forming neurons, just
as the timing of the thalidomide cases
predicted. The animal findings also pre-
dict that the Cerebellum must be affected
in the thalidomide cases-a hypothesis
that could be tested.
In a well-documented autopsy case
of autism, we have been able to demon-
strate a brain stem injury related to both
the thalidomide symptoms and the char-
acteristics of the animal model [Rodier et
al., 19961. The autopsy case has abnor-
malities of the facial and hypoglossal
nuclei, as well as a distinctive shortening
of the brain stem localized just caudal to
the facial nucleus. When we measured
the distances from one nucleus to another
in VPA-exposed rats, we found a signifi-
cant shortening in the same region in
each treated group [unpublished data].
Further studies of the cranial nerve nuclei
and brain stem distances in human cases
are needed to determine whether such
injuries are typical of all cases or only a
subset of the autistic population.
Unknowns
Even if the animal model we have
described and outlined in Figure 3 should
be a perfect representation of the CNS
abnormality underlying autism, the dis-
ease will remain mysterious until we
answer many basic questions. Knowing
how the initiating injury occurs does not
tell us the degree of injury required to
produce symptoms, or what parts of the
injury actually mediate the symptoms. At
this early stage of examining the model,
we already have recognized what appears
to be a secondary injury of the cerebel-
lum. What other parts of the brain have
been altered? For example, do the
VPA-exposed animals have increased
cell-packing density in the limbic system?
How is the brain stem wired in these
animals? One might expect changes in
the development of pathways when
early-forming cell groups are absent as
later-forming groups make their connec-
tions. What other toxic agents have the
potential to cause similar injuries? These
are a few of the questions that might be
addressed in an animal model. From
humans, we need many other kinds of
information. For example, are the thalido-
nlide cases representative of all cases of
autism? H o w does the genetic factor or
factors in the disease [reviewed in Piven
and Folstein, 19941 contribute to symp-
toms? What behavioral tasks can be
applied to animals to diagnose some as
“autistic” and others as “not autistic?”
SUMMARY
Some models of brain damage
work backward from the behavior to
hypotheses about the underlying injury,
but when information on the nature of
the injury is available, it is possible to
work forward from the injury to the
behavior or other characteristics of the
syndrome under study. In the case of
autism, the data from the thalidomide
cases specifi a type of injury and a
developmental stage when it occurs. The
injury indicated by the facts of these cases
is diffe-ent from the injuries one might
hypothesize from descriptions of autism’s
behavioral symptoms. However, our de-
scriptions of the symptoms of autism and
other brain damage syndromes include
interpretations that may or may not be
accurate. For example, the lack of facial
expression typically described as a defi-
ciency of social response o r conimunica-
tion is clearly due to lack of normal
innervation of the facial muscles in several
of the thalidomide cases and in our
autopsy case. The great advantage of
beginning with the facts of the injury is
that we need no interpretation to create
an animal model of what happened in the
thalidomide cases. The injury can be
reproduced in animals and the model
animals are robust and free of external
malformations. They have distinctive
changes in the cerebellum and shortening
of the medulla, paralleling human cases of
autism. This suggests that VPA-exposed
animals may serve as a model of the
disease. The value of the niodel will
depend on how well it provides new test
hypotheses for studies of human cases. W
ACKNOWLEDGMENTS
This research was supported by
research grants R01 NS24287 and R01
AAO8666 from NIH and R824758 from
EPA.
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