Neurotox Res (2011) 19:286–307
DOI 10.1007/s12640-010-9205-z
Critical Age Windows for Neurodevelopmental Psychiatric
Disorders: Evidence from Animal Models
Eva M. Marco • Simone Macrı̀ • Giovanni Laviola
Received: 11 November 2009 / Revised: 1 June 2010 / Accepted: 1 June 2010 / Published online: 7 July 2010
! Springer Science+Business Media, LLC 2010
Abstract A disruption in normal brain development has
been hypothesized to contribute to the aetiology of
major psychiatric disorders. According to the ‘double-hit’
hypothesis, mutant genes-based deviations, associated with
specific environmental insults during brain development,
may result in neurobehavioural disturbances. The existence
of age windows of vulnerability to environmental condi-
tions during brain maturation will be discussed, using as
examples a series of studies we have performed during the
last years. Major deviations from normative neurobehavi-
oural trajectories have been reported in animal models
following exposure to severe stress (either episodes of
maternal separation, deprivation or corticosterone supple-
mentation) early in infancy. Rodent models of difficult and/
or stressful pregnancies, including obstetric complications
(e.g. prenatal restrain stress or neonatal hypoxia) and ges-
tational exposure to infection (e.g prenatal immune chal-
lenge), have been associated with profound long-lasting
deficits in the offspring’s emotional and social behaviour,
and with immune and endocrine changes. More recently,
adolescence, characterized by elevated rates of brain plas-
ticity, has emerged as an additional period during which
sensitivity to environmental influence (either adverse or
stimulatory) is maximal. We have reported that both phar-
macological (methylphenidate) and environmental (physi-
cal or social enrichment) interventions can be used to
counteract the detrimental effects of earlier-origin devel-
opmental insults. Present findings indicate that these age
periods (i.e. prenatal stage, early infancy and adolescence)
E. M. Marco ! S. Macrı̀ ! G. Laviola (&)
Section of Behavioural Neuroscience, Department of Cell
Biology and Neurosciences, Istituto Superiore di Sanità, 00161
Rome, Italy
e-mail: giovanni.laviola@iss.it
123
do represent critical windows open to plastic changes and
might be susceptible to both adverse and supportive shaping
environmental forces. Taken together, age-related neuro-
plasticity might be considered not only as a risk factor for
psychopathology but also as a potent mechanism for com-
pensation. A better understanding of these critical periods
of brain development is a concern for public health and may
provide new insights into prevention strategies and into
novel therapeutic approaches in neuropsychiatry.
Keywords Adolescent ! Childhood ! Infant ! Stress !
Pregnancy ! Mental health
Introduction
Although genetic factors account for a significant propor-
tion of liability to psychiatric diseases, an increasing body
of evidence attests to a significant environmental contri-
bution, particularly during the developmental period.
Development represents a critical moment for shaping
adult behaviour and may set the stage to disease vulnera-
bility later in life. From the prenatal period through the first
years of life and until adolescence, the brain undergoes its
most rapid development, and is most open to the influence
of external experiences, for better or for worse. There is
now compelling evidence that suboptimal environments
during prenatal or early neonatal life alter development and
predispose the individual to lifelong health problems,
including susceptibility to mental illness (Cirulli et al.
2009; Laviola et al. 2009). Conversely, exposure to sup-
portive-enriching situations may favour the adaptation of
the developing brain to its environment, promoting better
coping strategies and, in certain cases, the possibility of
recovery from previous deleterious insults.
Neurotox Res (2011) 19:286–307
In this regard, animal models constitute a valuable tool for
the investigation of critical periods in human-brain devel-
opment. Animal studies can complement human studies,
whereby several parallelisms in age windows of suscepti-
bility to external environmental stimulus have been identi-
fied. Animal models are essential to answer key questions
relating to the neurobehavioural outcomes of developmental
manipulations. This approach offers the possibility of
revealing the complex and finely tuned mechanisms under-
lying developmental programming through in vivo and in
vitro studies at different time points. Herein, we will review
data from animal studies aimed at investigating the existence
of critical age windows of vulnerability to environmental
adversities (i.e. gestational period, early infancy and ado-
lescence). Neurobehavioural outcomes of rodent’s exposure
to both adverse and supportive environmental forces during
development will be described. Not only will these findings
be discussed from the point of view of psychopathology
resilience, but also in the prospective of therapeutic treat-
ment potentiality. A better understanding of critical periods
of brain development is a concern for public health and may
provide new insights into prevention strategies and into
novel therapeutic approaches in neuropsychiatry.
Critical Periods During Development
Developmental and congenital disorders of the brain can be
caused by environmental or genetic interference with nor-
mal brain development and result in neuropsychiatric dis-
ease. A critical period in developmental biology is
considered as a time in the early stages of an organism’s life
during which environmental stimulation is highly effective
in producing long-lasting changes. Critical periods were
first described as particular age periods during which
experience must occur for it to have an effect on subsequent
development and behaviour. In the best known example,
Konrad Lorenz proposed that young precocial birds had to
have exposure to conspecifics within hours of hatching to
develop adaptive filial attachments and appropriate later
mate preferences, the ‘imprinting’ phenomenon (Lorenz
1937/1957). Not long after Lorenz’s study on the critical
periods for the establishment of social behaviours in birds,
John Paul Scott (1962) reported a critical period for the
development of socialization in dogs and cats. Unless
encouraged to interact with humans at a particular early age,
certain breeds of dogs and cats would not be able to be
socialized to human interaction. Similarly, Harry Harlow
(1965) also reported a series of studies on the development
of social abilities in Rhesus monkeys that indicated that
social deprivation during the infant monkey’s first 3 months
had rather catastrophic long-term developmental conse-
quences. Later, Bowlby (1969) proposed a critical period
287
for the formation of an attachment relationship between the
mother and child which if disrupted resulted in the devel-
opment of adult psychopathology. The early postnatal
period appeared as a critical period for the formation of an
attachment relationship that would become the basis for all
future social interactions, and that would significantly affect
future emotional bonds. These studies led to the view that
critical periods were major phenomena in brain and
behavioural development (Michel and Tyler 2005); how-
ever, more in-depth studies of some of these critical periods
revealed that they were not as critical in terms of their
timing and specificity as previously supposed. Contempo-
raneously, Schneirla and Rosenblatt (1963) reported that the
development of species-typical social behaviour in cats
exhibited a cascading set of events in which the experiences
associated with each social event was critical for the cas-
cade. This last research argued against the notion of critical
periods in development and more for critical sequences for
the emergence of specific social skills and abilities (see
Michel and Tyler 2005 for historical review).
The notion of critical period nowadays coexists with that
of sensitive period and, despite conceptual differences, we
will use both terms indistinctly. Sensitive periods are asso-
ciated with neural plasticity (i.e. maturational events of
neurogenesis, differentiation and survival), a major salient
feature of the nervous system, particularly during brain
development—from the formation of the neural plate to the
adolescent period (Nowakowski and Hayes 1999; Acosta
et al. 2002; Andersen 2003; Butz et al. 2009)—although
neural plasticity has also been observed at adulthood
(Mackowiak et al. 2004). The architecture and physiology of
the brain develop throughout gestation and continue post-
natally through adolescent development, although disconti-
nuities in neurodevelopmental trajectories have been
described (Giedd 2008; Giedd et al. 2009). Insult at any point
during this period may result in aberrant neural structure or
function. Maturational processes occur at different times in
almost every brain region and neural circuit; the timing of
developmental events varies with neural substrate. There-
fore, critical periods might be specific for each brain region
or neurotransmitter systems depending on the specific vul-
nerability of each brain structure to outside influences (Rice
and Barone 2000; Johnson 2005; Andersen and Teicher
2008). Taken together, these critical periods during brain
development can be considered as the two sides of the same
coin. Exposure to adversity (i.e. stressful live events) might
enhance deleterious long-lasting outcomes, frequently
related to psychiatric diseases; conversely, exposure to
favourable situations (i.e. nurturant parenting and/or sup-
portive-enriching environment) may favour the adaptation
of the developing brain to its environment, promoting
improved coping strategies and, in certain cases, the possi-
bility of recovery from previous deleterious insults.
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Neurodevelopmental Hypothesis for Neuropsychiatric
Disorders
Disruptions in normal brain development may underlie the
pathogenesis of major mental disorders. Accordingly, a
‘neurodevelopmental hypotheses’ for neuropsychiatric
disorders was proposed in the last decades. This hypothesis
postulates that insults during brain maturation may lead to
the activation of pathologic neural circuits later in life
favouring the emergence of psychopathological symptoms
during adolescence or young adulthood. Yet, the broadest
data for this neurodevelopmental hypothesis have been
derived from studies conducted in schizophrenia and aut-
ism (see for review Fatemi and Folsom 2009). First, epi-
demiological studies suggest that processes early in
development (e.g. early-life abuse, neglect, pregnancy
stress or obstetric complications) are involved in the
pathogenesis of these mental disorders (Heim and
Nemeroff 2001; Cannon et al. 2002; Kendler et al. 2002;
Biederman 2005; Shea et al. 2005; Teicher et al. 2006;
Morgan and Fisher 2007; Talge et al. 2007; Beydoun and
Saftlas 2008; Mittal et al. 2008; Weinstock 2008). This
notion is further supported by the fact that many genes of
susceptibility for psychiatric disorders have key roles in
neurodevelopment: Neuroligins, Neureglin-1, Dysbindin,
Reelin, Disrupted-in-Schizophrenia-1 (DISC1) and
SHANK3, genes that are associated with synaptic plasticity
(Sawamura and Sawa 2006; O’Tuathaigh et al. 2007;
Fatemi and Folsom 2009). Moreover, linkage and associ-
ation studies appear to support the idea that environmental
factors affect causative gene clusters to precipitate a
pathological phenotype in susceptible individuals. That is,
interaction of developmental exposure to adversity with
individual genetic liabilities may predispose to psychopa-
thology throughout life (Agid et al. 1999; Heim et al.
2008); a theory postulated by Gottesman and Hanson
(2005) as the ‘double-hit’ hypothesis (Gottesman and
Hanson 2005). Emotional or physical stressful episodes
occurring early in life may act as ‘second hit’, whereby
they can increase susceptibility to psychiatric disorders in
naturally predisposed individuals (Laviola et al. 2009).
Whilst genetics play a key role in psychiatric disorders,
an increasing body of evidence supports the view that
environmental factors—particularly during specific phases
of development—concur to (dys)functional regulations.
Analysing gene–environment interactions in the liability to
adult neuropsychiatryc disorders remains far beyond the
scope of the present review. Herein, we will discuss
environmental risk factors likely to interfere with normal
brain development. Further understanding this neurode-
velopmental contribution to psychopathology will inform
public debate about the health effects of external insults
during individual’s growth but also open avenues in
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Neurotox Res (2011) 19:286–307
discerning how modulation of the early environment may
provide a valuable therapeutic tool for a variety of neuro-
psychiatric disorders.
Rodent Models of Early-Life Stress and Development
of Abnormal Neurobehavioural Phenotypes
In 1918, Sigmund Freud (1918) proposed that ‘every
neurosis in an adult is built upon a neurosis which has
occurred in his childhood’ thus suggesting that the adver-
sities encountered during the early stages of life impinge on
the adult phenotype. Several recent human studies support
this view. Specifically, a history of early-life abuse or
neglect appears to increase the risk of psychopathology
development at adulthood (Heim and Nemeroff 2001; Shea
et al. 2005; Teicher et al. 2006). Accordingly, parental loss
during childhood has been reported to significantly
increase the likelihood of developing major depression and
psychosis (Kendler et al. 2002; Morgan and Fisher 2007).
This idea roots its systematic basic neuroscience scientific
support in the pivotal studies of Harry Harlow (Harlow
et al. 1964), Otto Weininger (Weininger 1954) and Sey-
mour Levine (Levine 1957), who demonstrated that vari-
ations in the neonatal environment had effects on the
developing individual that persisted throughout the entire
lifespan. Specifically, they demonstrated that neonatal
maternal separation in both rodents and monkeys is suffi-
cient to persistently modify the development of those
behavioural and neuroendocrine systems involved in the
pathophysiology of human psychiatric disorders (e.g.
depression and anxiety). Nowadays, a consensus on the
critical role of the early environment in programming
neurobiological development has been achieved. Experi-
ences during the first periods of life influence behavioural,
emotional, cognitive and physiological properties at
adulthood.
Here, we aim to provide an integrative summary of the
long-term neuropathological consequences of environ-
mental insults during early developmental stages (i.e.
prenatal, neonatal and adolescence) by using rodent
models. In addition, we highlight how abnormalities in
neurotransmitter systems may provide a neural basis for
the emergence of specific forms of psychosis-related
behaviour. Animal models are valuable experimental tools
in the study of possible causal links between neurode-
velopmental disruption, specific neuronal dysfunctions
and distinct forms of abnormal behaviour. They are,
therefore, valuable in the exploration of specific brain and
behaviour relationships in complex multifactorial neuro-
psychiatric disorders that are likely to involve neuronal
disturbances beyond one single brain region and neuro-
transmitter system.
Neurotox Res (2011) 19:286–307
Perinatal Stress in Rodents: Methodology and Main
Results
Events occurring within the intrauterine or perinatal envi-
ronment may have significant consequences for the
development and function of physiological systems
throughout an individual’s lifespan. Here, we discuss the
existing literature on the negative effects of prenatal stress
(i.e. difficult and/or stressful pregnancies, including
obstetric complications) and gestational exposure to
infection (e.g prenatal immune challenge) in the psycho-
physiological homeostasis of the offspring.
Early Prenatal Immune Challenge
A consistent body of evidence based on human epidemi-
ological data supports the association between maternal
infection during pregnancy and higher risk of schizophre-
nia in the offspring (Patterson 2007, 2009; Boksa 2008 ;
Brown 2006). Similarly, an increasing number of experi-
mental studies in rats and mice demonstrate that prenatal
immune challenge is causally linked to the emergence of
major behavioural alterations in adulthood (for review
Meyer et al. 2009). Maternal exposure, during pregnancy,
to influenza, poliovirus, rubella, measles, varicella–zoster,
retrovirus (Yolken et al. 2000; Brown and Susser 2002; Shi
et al. 2003; Zuckerman et al. 2003) and bacterial agents
(Borrell et al. 2002; Fortier et al. 2004), has been associ-
ated with an increased risk for schizophrenia and/or autism
in the offspring. Actually, many of the functional deficits
induced by prenatal immune challenge in rats and mice
have been associated with critical phenotypes of schizo-
phrenia and autism. Impairments in prepulse and latent
inhibition response have been consistently reported fol-
lowing prenatal immune challenge; abnormalities in
working memory, exploration impairment and deficits in
social behaviour have also been observed (see Meyer et al.
2009 for review). Maternal immunological stimulation has
also been reported to induce long-term changes in multiple
neurotransmitter levels in the brains of adult offspring,
including dopamine and serotonin mainly within the limbic
system. The converging evidence from these studies sug-
gests a causal relationship between activation of the
immune system during pregnancy and altered behaviour in
the offspring. In rats, neonatal administration of proin-
flammatory cytokines (Tohmi et al. 2004) or leukaemia
inhibitory factors (Watanabe et al. 2004) induce psycho-
behavioural and/or cognitive impairments later in life.
Thus, it is likely that the maternal immune response, par-
ticularly involving the proinflammatory cytokines, may
interfere with normal foetal brain development and matu-
ration, and subsequently lead to structural and functional
deficits that compromise neurobehavioural functions at
289
adulthood (see Penner and Brown 2007; Moy and Nadler
2008; Patterson 2009; Meyer et al. 2009 for review).
Prenatal Restraint Stress
Clinical observations have enabled to delineate an associ-
ation between psychological stress during pregnancy and
impairments in emotional and cognitive development
among the offspring. Stress during pregnancy has been
shown to relate to increased incidence of anxiety, depres-
sion and attention deficits in children, together with major
neuro-behavioural disturbances like autism and schizo-
phrenia (Talge et al. 2007; Beydoun and Saftlas 2008;
Weinstock 2008). Moreover, maternal stress during preg-
nancy may not only increase risk for these disorders in
adulthood, but it may also facilitate illness onset and
comorbidity (Rice et al. 2007; Mill and Petronis 2008;
Mittal et al. 2008; Sarkar 2008). Neurobehavioural altera-
tions similar to those observed in humans can be investi-
gated in laboratory rodents and non-human primates
exposed to stress during pregnancy. In this regard, a great
variety of stressors have been used (for different examples
please refer Coe et al. 2002; Kofman 2002; Maccari et al.
2003; Koenig et al. 2005; Abe et al. 2007; Lee et al. 2007b;
Weinstock 2008). Our research has focused on the repeated
exposure to restraint stress during the last week of gestation
(Prenatal Restraint Stress, PRS). In particular, pregnant
dams were submitted to restrain immobilization under
bright light for 45 min three times a day during the last
week of gestation (from gestational day 11 to delivery)
(Morley-Fletcher et al. 2003b; Laviola et al. 2004b).
In our hands, adolescent male rats from stressed preg-
nancies displayed a prominent reduction in social play
behaviour when compared with animals raised under
standard conditions. However, no changes in environ-
mental exploratory parameters were observed (Morley-
Fletcher et al. 2003b). These results paralleled previous
literature showing that adolescent rats from disturbed
pregnancies were impaired in affiliative behaviour and
exhibited a reduced amount of age-typical rough-and-
tumble play (Ward and Stehm 1991; Koenig et al. 2005;
Lee et al. 2007b). Similarly, children born following
stressful pregnancies were less sociable than their peers
(Meijer 1985). Additionally, poor social interaction is
hallmark in psychiatric patients whose mothers experi-
enced severe psychological stress during pregnancy (Sus-
man 2006; Rice et al. 2009). Compared to control subjects
(offspring from non-stressed dams), adolescent rats from
disturbed pregnancies displayed increased anxiety-like
profile in an open field. Specifically, they showed a higher
latency to approach a novel object in this testing arena
(Laviola et al. 2004a, b). Latency to approach an unfa-
miliar object has been traditionally related to increased
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emotionality (Weinstock 2002). In line with these obser-
vations, adult PRS male rats showed increased anxiety-like
behaviour in the elevated plus-maze (EPM) (Darnaudery
and Maccari 2008; Zuena et al. 2008). Therefore, gesta-
tional stress may sensitize rats to the development of
hyper-anxiety, and perhaps depressive-like phenotypes,
whereby animals exposed to PRS also showed increased
immobility when evaluated in the forced swim test, a
behavioural profile sensitive to antidepressant drugs
(Morley-Fletcher et al. 2003a). Learning and attention
deficits have also been reported in the offspring of PRS
rats. Impairments in spatial learning in the water maze have
been observed in PRS male rats (Lemaire et al. 2000),
although gender differences exist with deficits being more
evident in female PRS rats (Zuena et al. 2008). Long-term
consequences of PRS significantly depend on the offspring
gender, on the behaviour being tested (see Table 1), and
most importantly on the intensity and timing of the
maternal stress (See Darnaudery and Maccari 2008;
Weinstock 2008 for review).
Dysfunction of the hypothalamus-pituitary-adrenal axis
(HPA) has been extensively observed following prenatal
stress. Specifically, adolescent and adult rats exposed to
stress during gestation show increased HPA axis reac-
tivity in terms of both ACTH and corticosterone
responses to stress. Surprisingly, at adulthood, no dif-
ferences in baseline corticosterone levels were observed,
whereas at adolescence, a significant reduction in basal
corticosterone levels was reported due to PRS (Morley-
Fletcher et al. 2003b; Laviola et al. 2004b). These results
then confirm the profile of impaired feedback inhibition
of HPA axis activity caused by maternal stress, and
evidenced in the offspring throughout the different pha-
ses of ontogenesis (see Maccari and Morley-Fletcher
2007 for review).
Furthermore, prenatal stress negatively affects the
immune system, and may possibly contribute to the mal-
adaptive immune responses to stress that occur later in life.
Moreover, psychosocial stress during early stages of pre-
and postnatal life may increase the vulnerability of infants
to the effects of immunotoxicants or immune-mediated
diseases (Bellinger et al. 2008, Merlot et al. 2008). In the
long term, PRS in rats produced important alterations in a
number of peripheral and central immunological parame-
ters, as previously described for several animal species
(Coe et al. 2002; Llorente et al. 2002; Tuchscherer et al.
2002; Gotz and Stefanski 2007). Specifically, a decrease in
blood cell populations devoted to immune competence,
especially the CD4 T lymphocytes and T4/T8 ratio were
observed in response to PRS (Laviola et al. 2004b). PRS
also produced an elevation in pro-inflammatory IL-1b
concentration, both in the spleen and the frontal cortex
(Laviola et al. 2004b). This increment in IL-1b production
among the PRS offspring resembles human studies sug-
gesting that stress and psychopathology can be indexed
by a hyper-secretion of pro-inflammatory cytokines,
although other intervening variables have to be considered
(Dabkowska and Rybakowski 1994; Elenkov and Chrousos
2002; Schiepers et al. 2005; Fan et al. 2007).
In summary, prenatal stress has important effects
on the programming of stress responsiveness, mainly
through the HPA axis. Animal studies also report critical
changes in the emotional, social and cognitive abilities of
the offspring, mediated, at least in part, by changes in
neural plasticity and brain neurotransmitter systems.
Research on animal models, i.e. prenatal restrain stress,
becomes of critical relevance for evaluating the psycho-
physiological outcomes of stressful pregnancies as well
as the possible underlying mechanisms. Heightened stress
during pregnancy increases plasma levels of cortisol and

Table 1 Long-term consequences of prenatal restrain stress (PRS) on adolescent and adult male rats
Paradigm Behavioural evaluation at adolescence References

Open field test
Social interaction test
Stress responsiveness
Paradigm
Increased latency to first approach a novel object (Laviola et al. 2004b)
Decreased social play behaviour in adolescent rats (Morley-Fletcher et al. 2003b)
No changes in environmental exploration (Laviola et al. 2004b)
Impairment in basal corticosterone levels (Laviola et al. 2004b)
Behavioural evaluation at adulthood References

Elevated plus-maze Increased anxiety-like behaviour (Zuena et al. 2008)

Forced-swimming test
Stress responsiveness
Water maze
Increased immobility, depressive-like behaviour (Morley-Fletcher et al. 2003a)
No changes in baseline corticosterone (Morley-Fletcher et al. 2003b)
An increased and prolonged corticosterone response to stress (Morley-Fletcher et al. 2003a)
Spatial learning deficits (Lemaire et al. 2000)
Spatial learning was not affected (Zuena et al. 2008)

PRS, pregnant Sprague-Dawley rats were submitted three times a day to 45 min sessions of restraint stress during the last week of gestation

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corticotrophin releasing hormone in the mother and foetus,
and may thus interfere with the required adaptive mecha-
nisms and lead to persistent dysregulation of the HPA axis
(Weinstock 2002; Lazinski et al. 2008). In animal studies,
PRS-induced impairments in behaviour and HPA axis
responsiveness are prevented by maternal adrenalectomy.
However, maternal injection of corticosterone only rever-
ses the increased anxiety and HPA alterations in the off-
spring (Weinstock 2008). Accordingly, the HPA axis offers
one hormonal pathway that could mediate some of the
reported changes associated to stress exposure during
pregnancy. Taken together, circulating concentrations of
maternal corticosteroids may constitute one of the critical
factors mediating the programming of foetal neuroana-
tomical and behavioural regulations. However, alterations
in the placental function due to stress during gestation
might be also considered as a critical factor mediating
deleterious outcomes in the offspring (O’Donnell et al.
2009).
Important neurobiological changes have been reported
following PRS in animal models. In the short-term (first
postnatal week, postnatal day, pnd 1–8), a decrease in brain
cell proliferation was observed in the hippocampus, together
with a reduction in brain-derived neurotrophic factor
(BDNF) content in the hippocampus (Van den Hove et al.
2006). Impairments in hippocampal neuroplasticity param-
eters have been also observed in the long-term, i.e. in adult
animals. In particular, PRS was associated with reduced
survival of newborn cells in the dentate gyrus, a reduced
activity of mGlu1/5 metabotropic glutamate receptors in the
ventral hippocampus, and an increase in BDNF levels within
the hippocampus (Zuena et al. 2008). Furthermore, a reduced
5-HT1A receptor binding in the ventral hippocampus has also
been described in adult animals exposed to PRS (Van den
Hove et al. 2006). These detrimental central changes, both in
neuroplasticity and neurotransmitter systems, may partly
explain the increased susceptibility of prenatally stressed
subjects to alterations analogous to those observed in psy-
chiatric, mostly depressive, patients. The postnatal envi-
ronment and the quality of maternal attention may also
critically determine the behaviour of the offspring (Wein-
stock 2008). In the recent years, great efforts have been
devoted to understand the underlying mechanisms of the
deleterious outcomes observed in the offspring of stressed
pregnancies (Weinstock 2002; Maccari et al. 2003;
Darnaudery and Maccari 2008; Weinstock 2008), however,
research in this field in still needed.
Neonatal Asphyxia
Early-life stress during the neonatal life frequently comes
in the form of obstetric complications. Obstetric compli-
cations, including perinatal asphyxia, are a major concern
291
for public health that occurs in approximately 1–6 per 1,000
live full-term births. Subtle neuropsychological deficits,
mostly impairments in attention and cognition impairments,
have been observed as a function of perinatal asphyxia
(Maneru et al. 2001; de Haan et al. 2006; van Handel et al.
2007; Angelini and Verklan 2009). Accumulating evidence
suggests that obstetric complications might be considered
as a predisposing risk factor in attention-deficit/hyper-
activity disorder (ADHD) (Biederman, 2005), and might
also contribute to the pathogenesis of schizophrenia
(Cannon et al. 2002; Mittal et al. 2008). More recently, an
association between obstetric complications and an
increased risk for autism, autism spectrum disorders and
bipolar disorder has also been suggested (Scott et al. 2006;
Kolevzon et al. 2007). Thus, obstetric complications may
interact with vulnerability genes to precipitate the onset of
several psychopathological conditions.
Much of our current understanding of the neurological
and neuropathological outcomes of obstetric complications
has evolved from animal studies. In line with human
observational data, studies using animal models indicate
motor and cognitive abilities to be significantly affected by
perinatal asphyxia (Rufini et al. 2001; Wakuda et al. 2008).
Accordingly, over the past decades, a great variety of
models involving hypoxia/ischaemia in foetal and newborn
non-human primates, lambs, piglets and rodents have been
developed (see for review Raju 1992; Painter 1995; Rufini
et al. 2001; Wakuda et al. 2008). As for laboratory rodents,
at least three different procedures have been employed. In
the first two models, rat pups undergo an episode of
‘hypoxia/ischemia’ (8% oxygen atmosphere plus occlusion
of the carotid artery) on postnatal day (pnd) 7, or caesarean
section followed by immersion of the uterus in a water bath
(‘neonatal anoxia’). Conversely, the third model consists in
the exposure of rat pups to a 100% nitrogen atmosphere
(‘neonatal asphyxia’) early after birth. Although there is no
consensus about which rodent-model best describes early
human-brain insults, among the diversity of hypoxia/
ischemia paradigms available in rats, several reasons made
us choose the ‘neonatal asphyxia’ model. In first place, it is
a non-invasive procedure, second, it eliminates the bias
associated with caesarean section (El-Khodor and Boksa
1997, 1998, 2001), and finally, this method allows the
offspring to be reared by their natural lactating dam, thus
avoiding fostering procedures. One obvious feature in this
model is that anoxic insults are delivered to independently
breathing animals, while hypoxia experienced by a human
foetus is dependent on maternal blood supply.
In a first series of experiments (see Table 2), newborn
rat pups (12–24 h after birth) underwent a single 30 min
exposure to 100% nitrogen (N2) atmosphere and short- and
long-term emotional alterations throughout infancy and
adolescence were evaluated. At pnd 15, corresponding to
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Table 2 Long-term consequences of neonatal asphyxia on adolescent and adult male rats
Paradigm Reference

Single episode of neonatal asphyxia (100% nitrogen (N2) atmosphere) for 30-min at postnatal day 1
Open field test (pnd 15) No changes in basal locomotion and exploration (Laviola et al. 2004a)
Reduced AMPH-induced hyperlocomotion Increased AMPH-induced
stereotypes
Novelty-seeking test (pnd 37–43) Stronger avoidance and inhibition profile. (Laviola et al. 2004a)
Social interaction test Decreased time spent in social behaviours, i.e. allogrooming and play- (Laviola et al. 2004a)
soliciting behaviours
Increased time devoted to object recognition
Repeated neonatal asphyxia (100% nitrogen (N2) atmosphere) for 20 min on alternate days during the first postnatal week (pnd 1 to 7)
Social interaction test (pnd 40) Higher involvement in investigative behaviours, i.e. anogenital sniffing (Adriani et al. 2006a, b)
Reduced play-soliciting behaviours, i.e. crawl over
Novelty-seeking test (pnd 43) Decreased locomotion (Adriani et al. 2006a, b)
Arousal profile
Impulsivity (pnd 68–78) Increased impulsive behaviour (Adriani et al. 2006a, b)
Elevated plus-maze (pnd 60) Higher emotional state (Casolini et al. 2005)
Morris water maze (pnd 75) No deficits in spatial learning (Casolini et al. 2005)
AMPH amphetamine

the infancy period, neonatal asphyxia did not change
locomotor and exploratory activity in the open field
(Laviola et al. 2004a). Conversely, in the novelty-seeking
test at adolescence, neonatal asphyxia reduced the prefer-
ence for the novel environment (Laviola et al. 2004a).
Considering that the behavioural profile of inhibition and
avoidance to novelty might represent a freezing response,
then, an episode of neonatal asphyxia may potentiate the
stressful and aversive component of novelty, at least in
some vulnerable individuals. Regarding social interactions,
rats exposed to neonatal asphyxia devoted less time to
social behaviours, i.e. allogrooming and play-soliciting
behaviours, in the absence of changes in the duration of
rough-and-tumble play. In contrast, neonatal asphyxia
increased the time spent in object exploration (Laviola
et al. 2004a). Thus, neonatal asphyxia appeared to reduce
the active search for social interaction, whereas it increased
the levels of environment-directed behaviours.
In a second series of experiments (see Table 2), we
modified the previously described ‘neonatal-asphyxia’
protocol (see above Laviola et al. 2004a) in order to mimic
repeated hypoxic complications occurring during the last
3 months of human pregnancy. Herein, asphyxia was
administered repeatedly (alternative days from pnd 1 to 7)
(Casolini et al. 2005; Adriani et al. 2006a) and behavioural
outcomes were evaluated throughout adolescence until
adulthood. During free-choice exposure to novelty, rats
exposed to neonatal asphyxia exhibited lower levels of
locomotion, as compared to controls. This behavioural
profile might underlay a reduced activation of the behav-
ioural component of the stress response as a consequence
of neonatal asphyxia (Adriani et al. 2006a). In agreement
with previous experiments (Laviola et al. 2004a) neonatal
asphyxia altered levels of social interaction in adolescent
rats. Additionally, in this study, neonatal asphyxia also
increased the frequency of ano-genital sniffing when
compared to controls (Adriani et al. 2006a), a spontaneous
behaviour that might lead to quite-aggressive social per-
formance (Alleva 1993; Terranova et al. 1998). At adult-
hood, rats exposed to this protocol of repeated anoxia
showed a higher emotional state when submitted to the
elevated plus-maze (Casolini et al. 2005). In spite of the
cognitive impairments frequently observed after neonatal
anoxia (Buwalda et al. 1995; Iuvone et al. 1996; Grojean
et al. 2003), our protocol did not affect spatial learning in
the water maze. Complementary measures of attention
were evaluated by means of the sudden silence test which
measures the level of attention in response to an abrupt
change of the environment. Animals from the neonatal
asphyxia group showed a substantially reduced immobility
when compared to control rats that responded with freezing
and immobility to a sudden interruption of the background
noise in the second day of testing (Casolini et al. 2005).
Taken together, neonatal asphyxia seems to affect emo-
tional response and attention processes, but not spatial
learning. The observations are in accordance with findings
from a different protocol of neonatal asphyxia (Caputa
et al. 2005). Impulsivity, defined as a reduced ability to
tolerate a delay of gratification, was studied—in an operant
behaviour task—by providing a choice between large-but-
delayed reinforcement versus smaller-and-immediate one
(Evenden and Ryan 1999). In the preference-delay curve, a

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Neurotox Res (2011) 19:286–307
significant difference in impulsivity was observed between
rats exposed to neonatal asphyxia and control rats (Adriani
et al. 2006a). The phenotype resulting from neonatal
asphyxia might suggest elevated levels of ‘motivational’
impulsivity, a dysfunction in reward processes often asso-
ciated with disinhibition and hyperactivity (Johansen et al.
2002), that could also explain the exaggerated anogenital
sniffing evaluated in the social interaction test. Changes in
the spontaneous drive towards the seeking of novelty and
social interactions with peers seem to be a consequence of
neonatal asphyxia.
Although no major structural damage was described
following the repeated protocol of neonatal asphyxia
(Casolini et al. 2005), relevant functional alterations
involving important neurotransmitter systems have been
reported. Compared to control animals, rat pups exposed to
neonatal asphyxia showed reduced amphetamine (AMPH)-
dependent locomotor hyperactivity and higher levels of
behavioural stereotypies (Laviola et al. 2004a). This result
suggests that an altered brain dopaminergic system may
underlie some of the behavioural effects associated with
perinatal asphyxia. Accordingly, changes in brain dopa-
mine (DA) systems have been previously related to peri-
natal asphyxia (Chen et al. 1997; Decker et al. 2003). A
decrease in the levels of the neurotrophin BDNF within the
striatum, but not in the fronto-parietal cortex, has also been
reported due to neonatal asphyxia (Laviola et al. 2004a),
and similar results were also found in the cerebellum
(Scheepens et al. 2003). One of the physiological roles of
BDNF is to enhance the functional activity of DA neurons
and to increase their synaptic performance (Dluzen et al.
2002; Goggi et al. 2002). Therefore, the reduced striatal
levels of BDNF present in adult rats neonatally exposed to
an anoxic atmosphere may suggest a diminished trophic
action onto DA neurons, and thus a poorer performance at
synaptic terminals (Adriani et al. 2006a). Serotonin
receptor levels were also profoundly altered as a conse-
quence of the early episodes of neonatal anoxia. Specifi-
cally, subjects submitted to neonatal asphyxia showed an
overexpression of 5-HT2A receptors in the striatum com-
pared to controls (Adriani et al. 2006a). Despite the evi-
dence for serotonergic modulation of dopaminergic
neurons through 5-HT2A receptors (Nocjar et al. 2002), this
interaction is currently unclear. Within the striatum,
important crosstalk between serotonergic and dopaminer-
gic systems might be of great relevance. Our results sup-
port a decrease in striatal BDNF levels as a result of
neonatal asphyxia (Laviola et al. 2004a), as well as an
overexpression of 5-HT2A receptors in the same brain
region. Therefore, changes in the expression of this sero-
tonergic receptor could be interpreted as a possible adap-
tation to the reduced BDNF input onto DA neurons
(Kostrzewa et al. 1998). Our research provides evidence for
293
long-lasting neurobehavioral deficits resulting from neo-
natal asphyxia in a rat model. Future studies should con-
sider additional variables that may modulate the impact of
asphyxia on behavioural function, and will forward the
goals of predicting long-term outcomes and understanding
the mechanisms by which it unfolds.
Early Neonatal Stress in Rodents: Methodology
and Main Results
Whilst the aforementioned studies investigated the effects
of perinatal stress on offspring development, many other
scientists addressed the effects of stress occurring during
early postnatal life. Hundreds of studies have addressed the
effects of neonatal mother–offspring separation of various
lengths on the development of the stress response system in
mammals. In particular, many authors observed, through
the disruption of usual mother–infant interactions in
rodents, changes in the neurobiology, physiology and
emotional behaviour of adult animals (Fenton 2001; Mea-
ney 2001; Pryce and Feldon 2003; Ellenbroek et al. 2005;
Macrı̀ and Wurbel 2006; Law et al. 2009). The neonatal
separation protocols aimed at interfering with normal dam–
pup interaction in rodents are generally applied during the
first 2 weeks of postnatal life. Regarding the effects of these
procedures, not only the duration of the separation protocol
is of great concern, but also the time window during which
such stress is applied. Finally, the time point at which the
evaluation of effects is undertaken results critical. Despite a
certain degree of discrepancies observed in literature—
dependent on the diversity of separation protocols—there is
a general consensus that adverse early experiences may
contribute to adult psychopathology (see for example
Levine 2005; Moffett et al. 2007). In resume, exposure to
stressful situations early in life may result in emotional
alterations, cognitive impairments and heightened fear
responses in adult rodents. Please find bellow several evi-
dences in that direction achieved in our laboratory.
Maternal Separation (MS)
Maternal separation (MS) in rodents involves repeated
3–6 h daily separations between postnatal day 1 and the
second or third week of life (Pryce and Feldon 2003 for a
review). These protocols induce a primary disturbance of
the mother–offspring interaction thereby mimicking a form
of disrupted attachment (Bowlby 1988) and infantile stress.
Several studies, performed in independent laboratories,
showed that maternal separation increases plasma levels of
adreno-cortico-tropic-hormone (ACTH) and corticosterone
in basal conditions and in response to restraint stress,
corticotropin-releasing-factor (CRF) in the cerebro-spinal-
fluid (CSF), and reduces glucocorticoid receptor density in
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294
the hippocampus (see e.g. Macrı̀ and Wurbel 2006). Spe-
cifically, we demonstrated that long periods of early
maternal separation in Lister-Hooded rats increased—in
adulthood—ACTH and corticosterone response to restrain
stress and behavioural fearfulness measured as the time
spent in the centre of an open field (Macrı̀ et al. 2004).
Maternal separation is also associated with reduced sero-
tonin reuptake transporter in the raphe nucleus and
increased behavioural immobility in the forced swim test
(Lee et al. 2007a) and the open field (Plotsky and Meaney
1993; Liu et al. 1997; Macrı̀ et al. 2004). Furthermore,
repeated mother–offspring separations have been shown to
reduce motivation to self-administer palatable food
rewards in rats (Matthews and Robbins 2003; Ruedi-
Bettschen et al. 2005). The latter has been traditionally
considered an index of anhedonia. Since separation-
dependent reduction in maternal care has been suggested to
potentially mediate these effects, we also studied absolute
levels of maternal care in MS and in non-handled (control)
dams. In contrast with our predictions, we observed that,
compared to non-handled conditions, maternal separation
increased absolute levels of maternal care (Macrı̀ et al.
2004). We, therefore, suggested that the effects of maternal
separation on mother–offspring interaction and adult off-
spring HPA regulation dissociate (Macrı̀ and Wurbel
2006). In order to address this possibility, we attempted to
selectively manipulate levels of active maternal care and
durations of mother–offspring separations in neonate rats.
Rat pups were exposed to different combinations of early
handling and MS from postnatal day (pnd) 2–10 using a
split-litter design. Maternal behaviour was recorded from
pnd 2 to 8 and behavioural and endocrine responses to
stress were studied in adult male offspring (Macrı̀ et al.
2008). Low levels of maternal care combined with long
mother–offspring separations increased HPA-reactivity
compared to both high maternal care combined with long
mother–offspring separations and low maternal care com-
bined with brief separations. These findings supported the
hypothesis that active maternal care and long mother–off-
spring separation act independently, and exert opposing
effects, on adult offspring’s HPA responses, but that
increased maternal care may buffer the adverse conse-
quences of long separations (Macrı̀ et al. 2008).
Since maternal care cannot be considered the unique
factor mediating the effects of neonatal stressors, we sug-
gested that circulating levels of corticosterone, presumably
rising in response to separation stress (D’Amato et al.
1992), to be transmitted to the offspring through lactation,
may be a candidate mediator of these responses (Macrı̀ and
Wurbel 2006). Indirect experimental evidence demon-
strated that corticosterone supplementation in the maternal
drinking water induced persistent alterations in stress and
fear responses and HPA regulation in the adult offspring
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Neurotox Res (2011) 19:286–307
(Catalani et al. 1993, 2000). Specifically, low doses of
corticosterone supplemented in the rat maternal drinking
water (200 mg/l) throughout the entire lactation period (i.e.
between birth and weaning) reduced anxiety-related
behavioural responses and HPA activation in response to
external stressors (Catalani et al. 1993, 2000). Thus, these
studies showed that exogenous corticosterone administered
to dams during lactation directly regulated stress and fear
responses in the adult offspring. We recently started to
investigate the possibility that elevated doses of cortico-
sterone may be used to up-regulate stress and fear
responses and HPA-reactivity as a potential model of early-
life stress (Macrı̀ et al. 2007a, 2009). Compared to
maternal separation protocols, corticosterone administra-
tion has several advantages: first, it allows a simulation of
the hormonal events occurring during stress while mini-
mizing the impact of uncontrollable confounds (e.g. effects
of uncontrolled handling procedures on dams and pups,
nest disturbance); second, it allows a fine-grain regulation
of the independent variable mimicking environmental
stress (corticosterone administration) both in terms of
quantity (adjustable doses) and in terms of timing. Based
on these considerations and on previous literature, we
hypothesized that whilst low doses of neonatal corticoste-
rone, presumably mimicking low levels of external stim-
ulation, may down-regulate later stress and fear responses
in the adult offspring (as in Catalani et al. 1993), high doses
of neonatal corticosterone, mimicking severe external
stress/adversity, may have opposite effects. In order to test
this hypothesis, we administered low (33 mg/l) and high
(100 mg/l1) doses of corticosterone to mouse dams during
the first week of lactation and then investigated the effects
of corticosterone administration in developing offspring
(Macrı̀ et al. 2007a, 2009). In line with our predictions, we
observed that elevated doses of neonatal corticosterone—
that were directly transmitted to the progeny through lac-
tation—reduced hippocampal BDNF levels and the number
of natural antibodies directed to dopamine transporter
(Macrı̀ et al. 2009). Furthermore, we observed that elevated
levels of neonatal corticosterone reduce absolute levels of
maternal care and increase HPA-reactivity to restraint
stress in adulthood (Macrı̀ et al. 2007b). Although pre-
liminary, these studies demonstrate that maternal cortico-
sterone can be directly translated to the progeny and that it
modulates the development of individual phenotype.
Additionally, these studies indirectly support the hypothe-
sis that external stressors perceived by the dam may exert
their effects on offspring development through increasing
1
Please note that daily water intake per kg of body weight is
approximately four-fold higher in mice than in rats. Therefore,
whereas 200 mg/l constitute a low dose of corticosterone supplemen-
tation in rats, 100 mg/l constitute a high corticosterone dose in mice.
Neurotox Res (2011) 19:286–307
circulating levels of corticosterone, which are in turn
transmitted to the progeny through lactation.
Although neonatal corticosterone directly adjusts stress
and fear responses in the developing individual, the bio-
logical mechanisms underlying these adaptations have
remained elusive. A valid candidate target is the epigenetic
regulation of glucocorticoid receptor expression (Weaver
et al. 2004). Specifically, Weaver et al. (2004) demon-
strated that particularly active maternal styles are associ-
ated with reduced methylation in the region encoding for
the glucocorticoid receptors. Such methylation status is
associated with increased gene expression (Weaver et al.
2004). Although no such link has been yet demonstrated
between corticosterone administration and gene expres-
sion, future studies may offer scientific support to this
hypothesis. In conclusion, several independent studies
support the hypothesis of a strong link between neonatal
adverse experiences and abnormal regulation of stress and
fear responses in the adult offspring (Laviola et al. 2008).
Maternal Deprivation (MD)
As an alternative to the aforementioned protocols, long-
lasting consequences of a prolonged single episode (24 h)
of maternal deprivation during the first two weeks of
postnatal life (postnatal day 9) have been also extensively
investigated (see e.g. Marco et al. 2009). This early-life
stress procedure provides a profound disruption of the
‘natural’ patterning of dam-offspring interaction in mice
and rats (Levine 2000; Meaney 2001). Maternal depriva-
tion has been reported to induce both short- and long-term
alterations changes in the stress reactivity system (Levine
et al. 1991; Suchecki and Tufik 1997; van Oers et al. 1997;
Lehmann et al. 2002; Schmidt et al. 2004), and conse-
quently has been related to the emergence of depressive
symptoms. Moreover, adult animals exposed to MD
showed impairments in the prepulse inhibition response
(Ellenbroek et al. 1998; Ellenbroek and Cools 2002), a
disrupted latent inhibition response (Ellenbroek and Cools
1995) and disturbances in both the auditory sensory gating
and startle habituation (Ellenbroek et al. 2004); impair-
ments that are commonly described among schizophrenic
patients. Moreover, increased sensitivity to apomorphine
(Ellenbroek and Cools 2002) and amphetamine (Zimmer-
berg and Shartrand 1992) have been described in rodents
following a single episode of maternal deprivation. Thus,
in line with the dopamine hypothesis of schizophrenia
(Guillin et al. 2007), an altered dopaminergic neurotrans-
mission has been suggested to mediate, at least in part,
some of the detrimental outcomes of maternal deprivation
(Ellenbroek et al. 2005). An alteration in glutamatergic
neurotransmission has been demonstrated to underlay, at
least in part, some of the behavioural anomalies induced by
295
early maternal deprivation (Roceri et al. 2002; Pickering
et al. 2006). More recently, changes within the endocan-
nabinoid system have also been reported. A short-term
elevation of hippocampal levels of the endogenous can-
nabinoid ligand, 2-arachidonylglycerol (2-AG) has been
described together with a reduction in the expression of
CB1R within the same brain area. In contrast, the expres-
sion of CB2R appears to be increased as a consequence of
the early maternal deprivation procedure (Llorente et al.
2008; Suárez et al. 2009). Moreover, an impaired immu-
nological function has been reported as a consequence of
the maternal deprivation procedure in the short-term, and
the detrimental outcomes persisted into adulthood. In par-
ticular, a diminished lymphocyte proliferative response to
mitogens in the lymphoid organs analysed (i.e. spleen,
thymus and axillary nodes), together with a deficient che-
motaxis index were observed as a long-term effect of
maternal deprivation in rats (Llorente et al. 2007).
Relevance of Animal Models for Biological Psychiatry
The animal models here presented are highly suitable for
the elucidation of the environmental contributions to
developmental brain and behavioural abnormalities rele-
vant to psychopathology. Present data provide further
evidence for the value of animal models in biological
psychiatry. Classically, changes at the level of the seroto-
nergic and the dopaminergic systems have been also
associated with schizophrenia (Abi-Dargham 2007; Guillin
et al. 2007) and depression (Lowry et al. 2008; Stein 2008;
Nemeroff and Owens 2009). Accordingly, the selected
animal models here described often exhibit anomalies
within these neurotransmitter systems. Moreover, these
animal models are also valuable tool for the investigation
of comorbid psychiatric disorders, given that schizophrenia
(and other neuropsychiatric disorders) is often associated
with a variety of certain aspects of affective psychopa-
thology, including depression and anxiety-related symp-
tomathology (Fenton 2001; Green et al. 2003; Braga et al.
2004).
In most of the environmental procedures previously
described, a dysregulation of the HPA axis is commonly
observed. An adequate function of this stress response
system appears to be essential for the well being of the
organism; in turn, impairments in its physiology can result
in potentially deleterious consequences such as altered
stress reactivity and inability to cope with stressful situa-
tions. Indeed, evidence suggests that multiple mood dis-
orders, including anxiety conditions and major depression,
are associated with chronic alterations in the circulating
levels and circadian rhythm of adrenal glucocorticoids in
the control of emotional reactivity (de Kloet 2003; Akil
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296
2005; de Kloet et al. 2007; Rozeboom et al. 2007). Fur-
thermore, HPA axis dysfunction may be a major biological
factor underlying the relationships between psychological
experience of stress and the development of psychosis,
particularly in the early phase of schizophrenia (Phillips
et al. 2006). Similarly, immunological abnormalities have
long been described in diverse psychiatric disorders,
including schizophrenia (Muller et al. 2000; Schwarz et al.
2001; Strous and Shoenfeld 2006). Accordingly, gluco-
corticoid levels have been given a critical role in coordi-
nating the immune system function (Marchetti et al. 2001).
Therefore, the endocrine and immunological consequences
of environmental insult during critical phases of brain
developmental (i.e. prenatal and early postnatal period)
may be strongly related. Consequently, these rodent mod-
els may provide a valuable tool for the evaluation of
endocrine and immunological interactions during brain
development, and for their influence on psychobiological
responses later in life.
Adolescence as a Sensitive Period to Developmental
Interference
Adolescence is the transition period between childhood and
adulthood, a stage of dramatic neuroendocrine and physical
changes. The concept of adolescence is overlapping with
that of puberty, but the two are not interchangeable. Pub-
erty is associated with the development of reproductive
function, and can thus be exactly defined in physiological
terms, while adolescence refers also to psychological and
social factors, and their limits are not always well defined.
Adolescence is characterized by psychological changes
that affect individual’s sense of identity, their self-con-
sciousness and their relationships with others. Compared
with adults, adolescents are more sociable, and display a
propensity to sensation-seeking and risk-taking behaviours
(La Greca et al. 2001; Kelley et al. 2004). On the one hand,
a prominent inner drive for sensation-seeking behaviour at
adolescence may have adaptive benefits with regard to the
development of independence and survival without paren-
tal protection. Actually, an adaptive role of adolescence
during hominid evolution has been proposed (Lockwood
et al. 2007). In general, increments in lifespan were
accompanied by an extension of the adolescent phase that
enabled a longer period for the exploration of new envi-
ronments thus allowing the emergence of innovative
behaviours. On the other hand, these increased levels of
exploration may lead to risk-taking behaviours, including
experiences with psychotropic drugs (Adriani and Laviola
2004; Schepis et al. 2008; Doremus-Fitzwater et al. 2009).
Since most of the psychobiological aspects of the tran-
sition from infancy into adulthood can be identified in most
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Neurotox Res (2011) 19:286–307
mammalian species, it is possible to study some of these
variables in animal models. In fact, adolescent rodents and
non-human primates display a behavioural repertoire sim-
ilar to that described for humans. Particularly, adolescent
animals exhibit increased levels of social play and affiliate
behaviours (Primus and Kellogg 1989; Vanderschuren
et al. 1997) as well as novelty-seeking and risk-taking
behaviours (Laviola et al. 1999; Spear 2000; Laviola et al.
2003; Adriani and Laviola 2004). Developmental discon-
tinuities in brain development have been hypothesized to
be in the bases of adolescent behaviour. Indeed, during
adolescence, a gradual loss of synapses together with a
strengthening of remaining synaptic connections has been
reported. Furthermore, a remarkable remodelling of corti-
cal and limbic circuitries is a biological hallmark of ado-
lescence, and major neurotransmitter systems continue
their maturational processes during adolescence (Blake-
more 2008; Casey et al. 2008; Giedd 2008). For more
detailed descriptions of the behavioural and neurobiologi-
cal features of adolescence please consult some specific
reviews, for e. g. Laviola et al. (1999), Spear (2000), and
Kelley et al. (2004).
Long-Lasting Neurobehavioural Effects of Adolescent
Exposure to Insult
Most of the studies have been developed in the field of drug
abuse and addiction since adolescence is an age in which
many risky behaviours including drug consumption and
smoking typically begin. Furthermore, adolescence repre-
sents a unique developmental stage for the effects of
chronic drug exposure. As far as we are concerned, long-
lasting neurobehavioural effects have been reported in
association with adolescent exposure to a vast amount of
substances (see for review: alcohol (Brown and Tapert
2004; McBride et al. 2005), nicotine (Slotkin et al. 2007;
Dwyer et al. 2009), cannabinoids (Rubino and Parolaro
2008), ecstasy or MDMA (Montoya et al. 2002; Piper
2007). In our laboratory, research has largely focused on
psychostimulants, i.e. nicotine and metilphenidate (MPH).
Nicotine administration during adolescence has been
extensively associated with important changes in the
expression of a great variety of central nervous system
receptors. A long-lasting up-regulation of nicotinic recep-
tors with a distinctly different regional pattern from that
seen in the adult has been described following adolescent
nicotine administration (Slotkin 2002; Adriani and Laviola
2004; Slotkin et al. 2007). Adolescent nicotine dose-
dependently down-regulated levels of AMPA GluR2/3
subunits in the striatum (Adriani and Laviola 2004). Sim-
ilarly, both cannabinoid CB1 receptor (CB1R) and
mu-opioid receptors (MOR) have been reported to modify
Neurotox Res (2011) 19:286–307
their expression following subchronic adolescent nicotine
administration (0.4 mg/kg daily from postnatal day 34–43).
Up-regulation and down-regulation of CB1Rs have been
reported for hippocampus and striatum, respectively. On
the contrary, striatal and hippocampal MOR expression
was reported to be down regulated in the long-term (Marco
et al. 2007). Mostly, only if nicotine was administered
during the adolescent period, were long-lasting neuro-
behavioural consequences reported. Thus, adolescence may
constitute a developmental period particularly vulnerable
to nicotine (Slotkin 2002; Adriani et al. 2003; Adriani and
Laviola 2004; Slotkin et al. 2007). These persistent long-
term effects of adolescent nicotine might be associated
with the behavioural alterations described following juve-
nile smoking. Mainly, impairments in reward pathways and
circuits involved in learning, memory and mood have been
observed in adolescent smokers (Slotkin 2002). Since
cigarette smoking during adolescence has been suggested
as a strong predictor of subsequent alcohol and other drug
use (Ellickson et al. 2001; Siqueira and Brook 2003;
Wetzels et al. 2003), a ‘gateway theory’ has been proposed
that describe a sequence and progression into addictive
substance use, from tobacco and alcohol to cannabis and
then to other illicit drugs like heroin and cocaine (Kandel
et al. 1992). In fact, cigarette smoking appears to be a
potent gateway drug for diverse illegal drugs (Lai et al.
2000). Other theories postulate that there is no clear tem-
poral sequence in the use of different substances, and that
individual phenotypes (affinity to sensation-seeking and
risky behaviours) may play a crucial role in drug abuse
among adolescents (for a review see Faeh et al. 2006).
Administration of methylphenidate (MPH, Ritalin") to
children with attention deficit hyperactivity disorder
(ADHD) is an elective therapy but raises concerns for
public health due to possible persistent neurobehavioural
alterations. Data from animal models indicate that expo-
sure to MPH during adolescence causes behavioural
changes enduring into adulthood. Increased anxiety-like
behaviours, enhanced plasma corticosterone levels, toge-
ther with decreased behavioural impulsivity and a depres-
sive-like phenotype have been described in adult animals
administered with MPH during adolescence (Bolanos et al.
2003; Carlezon et al. 2003; Adriani et al. 2007). Moreover,
motivational-choice behaviours are also affected. An
enhanced psychomotor responses to cocaine and increased
cocaine self-administration has been reported (Brandon
et al. 2001). More recently, a dissociation between damp-
ened place conditioning and enhanced locomotor sensiti-
zation induced by cocaine was reported in adolescent rats
administered with MPH (from pnd 30 to 46) when com-
pared to control animals (Adriani et al. 2006b). Further-
more, MPH-administered rats displayed altered processing
of incentive values and a modified flexibility/habit balance
297
(Adriani et al. 2006b). Given the critical involvement of the
striatum in regulating innate and acquired behavioural
habits, as well as natural reinforcement (Everitt and Rob-
bins 2005), the above-described behavioural alterations
may underlie fundamental plastic structural and functional
plastic changes in striatal circuits. Indeed, genome-wide
expression profiling, followed by RT-PCR validation,
showed a long-lasting up-regulation for the kainate 2
subunit of the ionotropic glutamate receptor (Grik2, also
known as GluR6) and for the serotonin (5-hydroxytrypta-
mine, 5-HT) receptor 7 (Htr7) within the striatum follow-
ing an adolescent exposure to MPH (Adriani et al. 2006b).
More recently, a marked increase of Htr7, mainly within
the nucleus accumbens, has been reported in the long-term
due to adolescent MPH administration. In parallel, behav-
ioural pharmacology studies have provided evidence for a
role of Htr7 receptors in the modulation of impulsive
behaviour (Leo et al. 2009). These results highlight the
need for further research to improve understanding of the
effects of psychostimulants (and other drugs of abuse) on
the developing nervous system and the potential-enduring
effects resulting from early-life drug exposure.
Chronic cannabinoid administration during adolescence
has been reported to cause persistent behavioural altera-
tions in relation to emotional responses (Biscaia et al.
2003; Macrı̀ and Laviola 2004; O’Shea et al. 2004), vul-
nerability to drug abuse (Ellgren et al. 2007) and cognitive
function (Schneider and Koch 2003). In humans, a debate
on the possible association between regular cannabis use
and psychotic disorders such as schizophrenia is still open
(see Di Forti et al. 2007; Hall and Degenhardt 2008;
Leweke and Koethe 2008 for review). There is increasing
evidence for the involvement of the endocannabinoid sys-
tem in psychotic disorders (Fritzsche 2001; Skosnik et al.
2001; Ujike and Morita 2004; Leweke and Koethe 2008).
A case–control study found a greater post-mortem density
of CB1R in the prefrontal cortex of schizophrenic diag-
nosed persons (Dean et al. 2001) and elevated levels of
anandamide (AEA) have been found in the cerebrospinal
fluid of non-medicated schizophrenic patients (Leweke
et al. 1999). Moreover, cannabis use has been reported to
increase positive symptoms and frequency of relapse and
hospitalization among schizophrenic patients (de Irala et al.
2005; Grech et al. 2005). However, present evidence is not
strong enough so as to confirm a causal relationship
between cannabis and psychosis. The maturational pro-
cesses that occur during adolescence are likely to confer
this age group a higher risk of suffering from adverse
consequences from cannabinoid exposure. Lasting conse-
quences on emotionality and cognition have been reported
in rodents (Schneider 2008). Moreover, these long-lasting
effects may increase not only the risk for neuropsychiatric
disorders, but also promote further illegal drug intake and
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298
increase the likelihood of cannabis dependence (Schneider
2008). Heavy cannabis consumption during adolescence is
a matter of public health concern so that efforts should be
invested in understanding the mental health risks of can-
nabis use, particularly among the juvenile population.
Despite the robust evidence pointing at adolescence as a
time of increased sensitivity to external influence, scarce
interest has been paid to investigating the disruption of
adolescent maturation by environmental insult. It has been
only in the recent years that some studies in experimental
animals have begun to address this data gap. In this regard,
social isolation—rearing rat pups from weaning in isola-
tion—is one of the most extensively studied procedures
that interfere with adolescent brain development causing
notable deleterious outcomes. In particular, social isolation
has been reported to cause behavioural, morphological and
neurochemical abnormalities that strongly resemble core
features of schizophrenia (see Fone and Porkess 2008 for
review). Similarly, exposure to stressful events during
adolescence, both in humans and rodent studies, has been
associated with the development of different psychopa-
thologies, including depression, anxiety and drug abuse
(Vidal et al. 2007; Andersen and Teicher 2008; Sondeijker
et al. 2008; Andersen and Teicher 2009; Greaves-Lord
et al. 2009). The exploration of the consequences of insults
during adolescent brain development is a matter of great
concern that should be explored in the future. Discovering
the mechanisms that trigger such deleterious outcomes will
improve our understanding of the aetiology of neurode-
velopmental psychiatric disorders, and may enable identi-
fication of longitudinal biomarkers of dysfunction that
might allow a predictive screening for novel compounds
with potential antipsychotic efficacy.
Critical Periods of Development: An Opportunity
for Intervention
Just as a critical period may be regarded as a phase during
which negative events may result in pathological regula-
tions in the developing individual, so also it may be con-
sidered a developmental stage during which positive events
may either facilitate future adaptations (Boyce and Ellis
2005; Ellis et al. 2005), or counteract the consequences of
negative events (Laviola et al. 2008). Within this realm,
several ‘beneficial’ treatments have been developed with
the aim of providing increased levels of stimulation to the
developing offspring. The ultimate goal was to demonstrate
that such stimulation might benefit the developing indi-
vidual and counteract the effects of an adverse perinatal
environment. A large body of literature has focused on two
developmental stages during which sensitivity to context is
maximal: early infancy and adolescence. Two treatments,
123
Neurotox Res (2011) 19:286–307
previously reported to reduce behavioural emotionality and
HPA responses to stressors (Weininger 1954; Weininger
et al. 1954; Levine et al. 1957), have been extensively used
in several studies: neonatal handling during the early stages
of neonatal life and environmental enrichment during
adolescence (Fernandez-Teruel et al. 1997; Laviola et al.
2004b; Nithianantharajah and Hannan 2006).
Early Handling
In 1954, Otto Weininger started to investigate a phenom-
enon that would intrigue a great number of scientists for at
least the following fifty years: the neonatal induction of
adult stress resilience. Weininger (1954) exposed neonate
albino rats to a treatment called ‘gentling’, consisting of
stroking the pup for 10-min a day during the first three
weeks of life. He observed that gentling reduced later
physiological damage and behavioural fearfulness in
adulthood. This study pioneered a series of studies in which
Levine and his collaborators (Levine 1957; Levine et al.
1957) showed that even daily maternal separations for as
little as 3 min per day during the first three weeks of life
had similar effects on later development. Specifically, adult
rats separated from the dams for 3-min per day during
lactation showed reduced adrenal gland weight 24 h after a
saline injection (Levine 1957). After these studies, many
other authors attempted to identify the factors mediating
such neonatal plasticity. Smotherman and Bell (1980)
proposed that the effects of the neonatal manipulations
were modulated by separation-induced changes in maternal
behaviour (‘maternal mediation hypothesis’). Maternal
care, in rats, occurs in the form of nursing bouts: dams
approach the pups, gather them underneath, lick them and
finally assume an ‘upright, crouching posture, in which (the
dam) stands over all or most of the pups with a pronounced
dorsal arch’ (Stern and Johnson 1990). Supporting evi-
dence to this possibility came from a number of studies
performed by Michael Meaney and collaborators, showing
that absolute levels of maternal care are increased fol-
lowing early handling (reviewed in Meaney 2001; Macrı̀
and Wurbel 2006). In particular, Liu et al. (Liu et al. 1997)
demonstrated that the reduced stress (ACTH and cortico-
sterone release following restraint stress) and fear respon-
ses shown by adult rats separated from their mothers for
brief periods (daily 15 min sessions, early handling, EH)
during the first week of life correlated with a handling-
induced increase in levels of active maternal care compared
to non-handled controls (Liu et al. 1997). Similarly,
spontaneously elevated levels of active maternal care in
non-handled dams were able to reduce adult offspring
neuroendocrine stress reactivity and behavioural fearful-
ness (Liu et al. 1997; but see also Francis and Meaney
1999). Thus, the early postnatal habitat mediates the
Neurotox Res (2011) 19:286–307
neuro-behavioural re-arrangements occurring between
infancy and adulthood, and these effects likely relate to the
amount of active maternal care received in infancy. Liu
et al. (1997) showed that the reduced endocrine stress
reactivity was dependent on a handling-induced increase in
glucocorticoid receptor density. Later studies (Meaney
et al. 2000) showed that early handling increased thyroid
hormone-mediated adenylyl cyclase activity and hippo-
campal cAMP levels through the activation of ascending
serotonergic pathways. Although, as already discussed (see
above), maternal care cannot explain all the effects asso-
ciated with manipulations of the neonatal environment,
several independent studies demonstrated that manipula-
tion-induced increases in maternal care are associated with
reduced stress and fear responses in adulthood (Liu et al.
1997; Macrı̀ et al. 2004). Under these conditions, early
handling has been adopted as a treatment to potentially
compensate for the negative consequences of prenatal
stress. For example, Lemaire et al. (2006) observed that
neonatal handling was able to counteract the effects of pre-
natal stress on hippocampal neurogenesis. Specifically,
adult rats born from dams exposed to three daily 45 min
restrain sessions during the last week of gestation showed
reduced hippocampal neurogenesis at 4 and 26 months.
These effects were abolished by neonatal handling,
whereby prenatally stressed rats exposed to handling pro-
cedures (10 min daily separations between birth and
weaning) showed analogous hippocampal cell proliferation
compared to controls (Lemaire et al. 2006).
Environmental Enrichment
Environmental enrichment is ‘a combination of complex
inanimate and social stimulation’ (Rosenzweig et al. 1978),
generally constituted by bigger housing cages, equipped
with a running wheel and few toys that are periodically
changed to stimulate animal curiosity and exploration
(Rosenzweig and Bennett 1996; Bezard et al. 1999; Laviola
et al. 2008). Enriched environments have powerful effects
on brain functions and structure, however, most studies
using enriched environments have focused, at the behav-
ioural level, on learning and memory functions, and, at the
cellular and molecular level, on the brain structures such
the hippocampus that are involved in memory and learning
(see van Praag et al. 2000 for review). Nonetheless, envi-
ronmental enrichment also directly interacts with those
systems that are generally impaired by early-life stressful
procedures. Thus, in rats, environmental enrichment redu-
ces emotionality in the open field and elevated plus-maze,
reduces ACTH and corticosterone response to various
stressors (electric shock and 20 min restraint), and
increases glucocorticoid receptor expression, 5-HT and
5-HIAA levels in the hippocampus (e.g. Fernandez-Teruel
299
et al. 2002a; Fox et al. 2006). Based on these findings,
similarly to what has been previously discussed for early
handling, several authors tested whether environmental
enrichment effects were powerful enough to counteract the
effects of early stress (Escorihuela et al. 1993; Francis et al.
2002; Morley-Fletcher et al. 2003b; Laviola et al. 2004b).
In line with the predictions, Francis et al. demonstrated that
access to an enriched environment reversed the anxiogenic
effects of maternal separation. Specifically, while maternal
separation reduced the time spent in the centre of an open
field, the propensity to forage in a novel environment and
increased corticosterone release following a stressor,
environmental enrichment reversed these effects (Francis
et al. 2002). Likewise, environmental enrichment has been
reported to counteract the effect of prenatal stress. We
already discussed the effects of prenatal stress on HPA
regulation, social behaviour and response to immune-sup-
pressive drug challenge (see above Morley-Fletcher et al.
2003b; Laviola et al. 2004b)). In line with the predicted
compensatory effect of environmental enrichment, we also
observed that prenatally stressed rats, kept in enriched
cages throughout adolescence, showed indistinguishable
corticosterone response to restraint and reactivity to an
immunosuppressive agent compared to standard reared
non-stressed individuals (Laviola et al. 2004b). The same
compensatory effects of environmental enrichment were
observed on playful behaviour (Morley-Fletcher et al.
2003), and in the 5-HT1A receptor in the dorsal hippo-
campus (e.g. Rasmuson et al. 1998). Finally, environmental
enrichment has also been reported to rescue animals from
the deleterious effects of neonatal asphyxia. Reduced
impulse control associated with neonatal asphyxia was
reverted by being reared in an enriched environment (pnd
21–35) (Adriani et al. 2006a). An increase in the striatal
expression of 5-HT2A receptor was proposed to be in the
bases of the elevated impulsive behaviour displayed by the
animals exposed to neonatal asphyxia. Accordingly, envi-
ronmental enrichment also counteracted this effect (Adri-
ani et al. 2006a). Some other studies further support the
protective effect of environmental enrichment as regards
detrimental outcomes of neonatal asphyxia. Being housed
in an enriched environment (from pnd 8–30) appeared as
an effective strategy for preventing some cognitive deficits
caused by neonatal hypoxia–ischemia (Pereira et al. 2008,
2009).
In conclusion, a large body of experimental evidence
supports the view that stimulating environments during life
periods of elevated brain plasticity may counteract the
negative effects of adverse pre- and post-natal stress. Early
handling and environmental enrichment are the most com-
monly used methods of providing supplementary environ-
mental stimulation in rodent models. Great efforts have
been made to understand the mechanisms that underlie such
123
300
‘protective’ effects (see Fernandez-Teruel et al. 2002b for
review). Very recently, a second cluster for the DSM-IV and
ICD-10 has been considered, namely neurodevelopmental
disorders (Andrews et al. 2009). This fact further emphasizes
the urgent need for understanding developmental program-
ming as well as the cellular and molecular processes that
underlie the environmental manipulations that ‘protect’ from
resilience to psychopathology.
Conclusion and Final Remarks
There is a great concern in understanding the aetiopatho-
genesis of behavioural abnormalities that characterize
multiple neuropsychiatric disorders. Taken together, our
data support the hypothesis of disrupted neural develop-
ment in diverse psychiatric disorders. Based on the epi-
demiological association between exposure to insult during
critical age periods and enhanced risk of neurodevelop-
mental brain disorders in the offspring, a number of rodent
models (only some examples presented here) have since
been established to study this link on an experimental
basis. These models prove to be valid experimental strat-
egies to test the hypothesis of causality and biological
plausibility of human epidemiological findings. Indeed, the
exposure of developing rats and mice to adverse environ-
mental conditions produces reproducible, long-term chan-
ges including: neophobia, increased emotionality,
depressive-like behaviours, impaired response to stressful
events and cognitive deficits, neurobehavioural abnormal-
ities that strongly resemble core features of diverse neu-
ropsychiatric disorders. Unravelling the mechanisms that
trigger these sequels will improve our knowledge of the
aetiology of neurodevelopmental psychiatric disorders,
enable identification of longitudinal biomarkers of dys-
function and allow the identification of new windows of
opportunity for the development of novel therapeutic
interventions, opening the possibility of pharmacological
and/or environmental approaches.
The existence of discontinuities in brain development,
including periods of remarkable neuroplasticity, might be
considered as a negative feature whereby adverse stressful
episodes might predispose to the emergence of psychopa-
thologies later in life. Yet, being raised under enriching/
stimulating conditions offers a wide range of possibilities
for early intervention and positive prognosis. Genders may
differ in the sensitivity of developing brain areas to
external influences, and to the putative mediating mecha-
nisms (e.g. stress hormones). Possible sexual dimorphisms
should be systematically studied in future clinical research
and animal studies. Only in the recent years have animal
studies incorporated both males and females, and signifi-
cant differences have been reported (Weinstock 2007;
123
Neurotox Res (2011) 19:286–307
Zuena et al. 2008). However, several gaps in the present
knowledge can be pointed out, especially concerning the
boundaries of identified time windows (if any), the indi-
vidual differences in developmental brain sensitivity, and
whether compensation mechanisms occurring at early
stages might induce a shift in physiological set points that
may persist into adulthood.
Acknowledgements Supported by the collaborative program NIH–
ISS 0F14 and by FP6-Program ERARE-EuroRETT Network (to GL).
EM Marco has been supported by a post-doctoral fellowship from the
Spanish ‘Ministerio de Ciencia e Innovación (MCINN)’, and is now
at the Departamento de Fisiologı́a (Fisiologı́a Animal II) (planta 13,
despacho 16) Facultad de CC. Biológicas Universidad Complutense
de Madrid, Madrid (Spain). S Macrı̀ has been supported by a NAR-
SAD young investigator award. G.L. and S.M. acknowledge the
support by the project ‘ECS-EMOTION’ from the Department of
Antidrug Policies c/o Presidency of the Council of Ministers, Italy.
We would also like to acknowledge our expert colleagues that con-
tributed to the original research data presented: W. Adriani, S. Mor-
ley-Fletcher, M. Rea and L. Aloe.
References
Abe H, Hidaka N, Kawagoe C, Odagiri K, Watanabe Y, Ikeda T,
Ishizuka Y, Hashiguchi H, Takeda R, Nishimori T, Ishida Y
(2007) Prenatal psychological stress causes higher emotionality,
depression-like behavior, and elevated activity in the hypotha-
lamo-pituitary-adrenal axis. Neurosci Res 59:145–151
Abi-Dargham A (2007) Alterations of serotonin transmission in
schizophrenia. Int Rev Neurobiol 78:133–164
Acosta M, Gallo V, Batshaw ML (2002) Brain development and the
ontogeny of developmental disabilities. Adv Pediatr 49:1–57
Adriani W, Laviola G (2004) Windows of vulnerability to psycho-
pathology and therapeutic strategy in the adolescent rodent
model. Behav Pharmacol 15:341–352
Adriani W, Spijker S, Deroche-Gamonet V, Laviola G, Le Moal M,
Smit AB, Piazza PV (2003) Evidence for enhanced neurobe-
havioral vulnerability to nicotine during periadolescence in rats.
J Neurosci 23:4712–4716
Adriani W, Giannakopoulou D, Bokulic Z, Jernej B, Alleva E,
Laviola G (2006a) Response to novelty, social and self-control
behaviors, in rats exposed to neonatal anoxia: modulatory effects
of an enriched environment. Psychopharmacology (Berl)
184:155–165
Adriani W, Leo D, Greco D, Rea M, di Porzio U, Laviola G, Perrone-
Capano C (2006b) Methylphenidate administration to adolescent
rats determines plastic changes on reward-related behavior and
striatal gene expression. Neuropsychopharmacology 31:1946–
1956
Adriani W, Canese R, Podo F, Laviola G (2007) 1H MRS-detectable
metabolic brain changes and reduced impulsive behavior in adult
rats exposed to methylphenidate during adolescence. Neurotox-
icol Teratol 29:116–125
Agid O, Shapira B, Zislin J, Ritsner M, Hanin B, Murad H, Troudart
T, Bloch M, Heresco-Levy U, Lerer B (1999) Environment and
vulnerability to major psychiatric illness: a case control study of
early parental loss in major depression, bipolar disorder and
schizophrenia. Mol Psychiatr 4:163–172
Akil H (2005) Stressed and depressed. Nat Med 11:116–118
Alleva E (1993) Assessment of aggressive behavior in rodents.
Methods Neurosci 14:111–137
Neurotox Res (2011) 19:286–307
Andersen SL (2003) Trajectories of brain development: point of
vulnerability or window of opportunity? Neurosci Biobehav Rev
27:3–18
Andersen SL, Teicher MH (2008) Stress, sensitive periods and
maturational events in adolescent depression. Trends Neurosci
31:183–191
Andersen SL, Teicher MH (2009) Desperately driven and no brakes:
developmental stress exposure and subsequent risk for substance
abuse. Neurosci Biobehav R 33:516–524
Andrews G, Pine DS, Hobbs MJ, Anderson TM, Sunderland M (2009)
Neurodevelopmental disorders: cluster 2 of the proposed meta-
structure for DSM-V and ICD-11. Psychol Med 1–11
Angelini DJ, Verklan MT (2009) Complications during the intrapar-
tum and newborn period. J Perinatal Neonatal Nurs 23:1–2
Bellinger DL, Lubahn C, Lorton D (2008) Maternal and early life
stress effects on immune function: relevance to immunotoxicol-
ogy. J Immunotoxicol 5:419–444
Beydoun H, Saftlas AF (2008) Physical and mental health outcomes
of prenatal maternal stress in human and animal studies: a review
of recent evidence. Paediatr Perinatal Epidemiol 22:438–466
Bezard E, Gross CE, Fournier MC, Dovero S, Bloch B, Jaber M
(1999) Absence of MPTP-induced neuronal death in mice
lacking the dopamine transporter. Exp Neurol 155:268–273
Biederman J (2005) Attention-deficit/hyperactivity disorder: a selec-
tive overview. Biol Psychiatr 57:1215–1220
Biscaia M, Marin S, Fernandez B, Marco EM, Rubio M, Guaza C,
Ambrosio E, Viveros MP (2003) Chronic treatment with CP 55,
940 during the peri-adolescent period differentially affects the
behavioural responses of male and female rats in adulthood.
Psychopharmacology (Berl) 170:301–308
Blakemore SJ (2008) The social brain in adolescence. Nat Rev
Neurosci 9:267–277
Boksa P (2008) Maternal infection during pregnancy and schizophre-
nia. J Psychiatry Neurosci 33:183–185
Bolanos CA, Barrot M, Berton O, Wallace-Black D, Nestler EJ
(2003) Methylphenidate treatment during pre- and periadoles-
cence alters behavioral responses to emotional stimuli at
adulthood. Biol Psychiatr 54:1317–1329
Borrell J, Vela JM, Arevalo-Martin A, Molina-Holgado E, Guaza C
(2002) Prenatal immune challenge disrupts sensorimotor gating
in adult rats. Implications for the etiopathogenesis of schizo-
phrenia. Neuropsychopharmacology 26:204–215
Bowlby J (1969) Attachment and loss, Attachment. Basic Books, New
York
Bowlby J (1988) Developmental psychiatry comes of age. Am J
Psychiatr 145:1–10
Boyce WT, Ellis BJ (2005) Biological sensitivity to context: I. An
evolutionary-developmental theory of the origins and functions
of stress reactivity. Dev Psychopathol 17:271–301
Braga RJ, Petrides G, Figueira I (2004) Anxiety disorders in
schizophrenia. Compr Psychiatr 45:460–468
Brandon CL, Marinelli M, Baker LK, White FJ (2001) Enhanced
reactivity and vulnerability to cocaine following methylpheni-
date treatment in adolescent rats. Neuropsychopharmacology
25:651–661
Brown AS (2006) Prenatal infection as a risk factor for schizophrenia.
Schizophr Bull 32:200–202
Brown AS, Susser ES (2002) In utero infection and adult schizo-
phrenia. Ment Retard Dev Disabil Res Rev 8:51–57
Brown SA, Tapert SF (2004) Adolescence and the trajectory of alcohol
use: basic to clinical studies. Ann NY Acad Sci 1021:234–244
Butz M, Worgotter F, van Ooyen A (2009) Activity-dependent
structural plasticity. Brain Res Rev 60:287–305
Buwalda B, Nyakas C, Vosselman HJ, Luiten PG (1995) Effects of
early postnatal anoxia on adult learning and emotion in rats.
Behav Brain Res 67:85–90
301
Cannon M, Jones PB, Murray RM (2002) Obstetric complications and
schizophrenia: historical and meta-analytic review. Am J
Psychiatr 159:1080–1092
Caputa M, Rogalska J, Wentowska K, Nowakowska A (2005)
Perinatal asphyxia, hyperthermia and hyperferremia as factors
inducing behavioural disturbances in adulthood: a rat model.
Behav Brain Res 163:246–256
Carlezon WA Jr, Mague SD, Andersen SL (2003) Enduring
behavioral effects of early exposure to methylphenidate in rats.
Biol Psychiatr 54:1330–1337
Casey BJ, Jones RM, Hare TA (2008) The adolescent brain. Ann NY
Acad Sci 1124:111–126
Casolini P, Zuena AR, Cinque C, Matteucci P, Alema GS, Adriani W,
Carpinelli G, Santoro F, Alleva E, Bosco P, Nicoletti F, Laviola
G, Catalani A (2005) Sub-neurotoxic neonatal anoxia induces
subtle behavioural changes and specific abnormalities in brain
group-I metabotropic glutamate receptors in rats. J Neurochem
95:137–145
Catalani A, Marinelli M, Scaccianoce S, Nicolai R, Muscolo LA,
Porcu A, Koranyi L, Piazza PV, Angelucci L (1993) Progeny of
mothers drinking corticosterone during lactation has lower
stress-induced corticosterone secretion and better cognitive
performance. Brain Res 624:209–215
Catalani A, Casolini P, Scaccianoce S, Patacchioli FR, Spinozzi P,
Angelucci L (2000) Maternal corticosterone during lactation
permanently affects brain corticosteroid receptors, stress
response and behaviour in rat progeny. Neuroscience 100:319–
325
Chen Y, Hillefors-Berglund M, Herrera-Marschitz M, Bjelke B,
Gross J, Andersson K, von Euler G (1997) Perinatal asphyxia
induces long-term changes in dopamine D1, D2, and D3 receptor
binding in the rat brain. Exp Neurol 146:74–80
Cirulli F, Francia N, Berry A, Aloe L, Alleva E, Suomi SJ (2009)
Early life stress as a risk factor for mental health: role of
neurotrophins from rodents to non-human primates. Neurosci
Biobehav Rev 33:573–585
Coe CL, Kramer M, Kirschbaum C, Netter P, Fuchs E (2002) Prenatal
stress diminishes the cytokine response of leukocytes to
endotoxin stimulation in juvenile rhesus monkeys. J Clin
Endocrinol Metab 87:675–681
D’Amato FR, Cabib S, Puglisi-Allegra S, Patacchioli FR, Cigliana G,
Maccari S, Angelucci L (1992) Effects of acute and repeated
exposure to stress on the hypothalamo-pituitary-adrenocortical
activity in mice during postnatal development. Horm Behav
26:474–485
Dabkowska M, Rybakowski J (1994) Stress, depression and schizo-
phrenia in view of psychoimmunology. Psychiatr Pol 28:23–32
Darnaudery M, Maccari S (2008) Epigenetic programming of the
stress response in male and female rats by prenatal restraint
stress. Brain Res Rev 57:571–585
de Haan M, Wyatt JS, Roth S, Vargha-Khadem F, Gadian D, Mishkin
M (2006) Brain and cognitive-behavioural development after
asphyxia at term birth. Dev Sci 9:350–358
de Irala J, Ruiz-Canela M, Martinez-Gonzalez MA (2005) Causal
relationship between cannabis use and psychotic symptoms or
depression. Should we wait and see? A public health perspective.
Med Sci Monit 11:RA355–RA358
de Kloet ER (2003) Hormones, brain and stress. Endocr Regul 37:51–
68
de Kloet ER, Derijk RH, Meijer OC (2007) Therapy insight: is there
an imbalanced response of mineralocorticoid and glucocorticoid
receptors in depression? Nat Clin Pract Endocrinol Metab 3:168–
179
Dean B, Sundram S, Bradbury R, Scarr E, Copolov D (2001) Studies
on [3H]CP-55940 binding in the human central nervous system:
regional specific changes in density of cannabinoid-1 receptors
123
302
associated with schizophrenia and cannabis use. Neuroscience
103:9–15
Decker MJ, Hue GE, Caudle WM, Miller GW, Keating GL, Rye DB
(2003) Episodic neonatal hypoxia evokes executive dysfunction
and regionally specific alterations in markers of dopamine
signaling. Neuroscience 117:417–425
Di Forti M, Morrison PD, Butt A, Murray RM (2007) Cannabis use
and psychiatric and cognitive disorders: the chicken or the egg?
Curr Opin Psychiatr 20:228–234
Dluzen DE, Anderson LI, McDermott JL, Kucera J, Walro JM (2002)
Striatal dopamine output is compromised within ?/- BDNF
mice. Synapse 43:112–117
Doremus-Fitzwater TL, Varlinskaya EI Spear LP (2009) Motivational
systems in adolescence: Possible implications for age differences
in substance abuse and other risk-taking behaviors. Brain Cogn
72:114–123
Dwyer JB, McQuown SC, Leslie FM (2009) The dynamic effects of
nicotine on the developing brain. Pharmacol Therapeut 122:125–
139
Elenkov IJ, Chrousos GP (2002) Stress hormones, proinflammatory
and antiinflammatory cytokines, and autoimmunity. Ann N Y
Acad Sci 966:290–303
El-Khodor BF, Boksa P (1997) Long-term reciprocal changes in
dopamine levels in prefrontal cortex versus nucleus accumbens
in rats born by Caesarean section compared to vaginal birth. Exp
Neurol 145:118–129
El-Khodor BF, Boksa P (1998) Birth insult increases amphetamine-
induced behavioral responses in the adult rat. Neuroscience
87:893–904
El-Khodor B, Boksa P (2001) Caesarean section birth produces long
term changes in dopamine D1 receptors and in stress-induced
regulation of D3 and D4 receptors in the rat brain. Neuropsy-
chopharmacology 25:423–439
Ellenbroek BA, Cools AR (1995) Maternal separation reduces latent
inhibition in the conditioned taste aversion paradigm. Neurosci
Res Commun 17:27–33
Ellenbroek BA, Cools AR (2002) Early maternal deprivation and
prepulse inhibition: the role of the postdeprivation environment.
Pharmacol Biochem Behav 73:177–184
Ellenbroek BA, van den Kroonenberg PT, Cools AR (1998) The
effects of an early stressful life event on sensorimotor gating in
adult rats. Schizophr Res 30:251–260
Ellenbroek BA, de Bruin NM, van Den Kroonenburg PT, van
Luijtelaar EL, Cools AR (2004) The effects of early maternal
deprivation on auditory information processing in adult Wistar
rats. Biol Psychiatr 55:701–707
Ellenbroek BA, Derks N, Park HJ (2005) Early maternal deprivation
retards neurodevelopment in Wistar rats. Stress 8:247–257
Ellgren M, Spano SM, Hurd YL (2007) Adolescent cannabis exposure
alters opiate intake and opioid limbic neuronal populations in
adult rats. Neuropsychopharmacology 32:607–615
Ellickson PL, Tucker JS, Klein DJ (2001) High-risk behaviors
associated with early smoking: results from a 5-year follow-up. J
Adolesc Health 28:465–473
Ellis BJ, Essex MJ, Boyce WT (2005) Biological sensitivity to
context: II. Empirical explorations of an evolutionary-develop-
mental theory. Dev Psychopathol 17:303–328
Escorihuela RM, Fernandez-Teruel A, Zapata A, Nunez JF, Tobena A
(1993) Flumazenil prevents the anxiolytic effects of diazepam,
alprazolam and adinazolam on the early acquisition of two-way
active avoidance. Pharmacol Res 28:53–58
Evenden JL, Ryan CN (1999) The pharmacology of impulsive
behaviour in rats VI: the effects of ethanol and selective
serotonergic drugs on response choice with varying delays of
reinforcement. Psychopharmacology (Berl) 146:413–421
123
Neurotox Res (2011) 19:286–307
Everitt BJ, Robbins TW (2005) Neural systems of reinforcement for
drug addiction: from actions to habits to compulsion. Nat
Neurosci 8:1481–1489
Faeh D, Viswanathan B, Chiolero A, Warren W, Bovet P (2006)
Clustering of smoking, alcohol drinking and cannabis use in
adolescents in a rapidly developing country. BMC Public Health
6:169
Fan X, Goff DC, Henderson DC (2007) Inflammation and schizo-
phrenia. Expert Rev Neurother 7:789–796
Fatemi SH, Folsom TD (2009) The neurodevelopmental hypothesis of
schizophrenia, revisited. Schizophr Bull 35:528–548
Fenton WS (2001) Comorbid conditions in schizophrenia. Curr Opin
Psychiatr 14:17–23
Fernandez-Teruel A, Escorihuela RM, Castellano B, Gonzalez B,
Tobena A (1997) Neonatal handling and environmental enrich-
ment effects on emotionality, novelty/reward seeking, and age-
related cognitive and hippocampal impairments: focus on the
Roman rat lines. Behav Genet 27:513–526
Fernandez-Teruel A, Driscoll P, Gil L, Aguilar R, Tobena A,
Escorihuela RM (2002a) Enduring effects of environmental
enrichment on novelty seeking, saccharin and ethanol intake in
two rat lines (RHA/Verh and RLA/Verh) differing in incentive-
seeking behavior. Pharmacol Biochem Behav 73:225–231
Fernandez-Teruel A, Gimenez-Llort L, Escorihuela RM, Gil L,
Aguilar R, Steimer T, Tobena A (2002b) Early-life handling
stimulation and environmental enrichment: are some of their
effects mediated by similar neural mechanisms? Pharmacol
Biochem Behav 73:233–245
Fone KC, Porkess MV (2008) Behavioural and neurochemical effects
of post-weaning social isolation in rodents-relevance to devel-
opmental neuropsychiatric disorders. Neurosci Biobehav R
32:1087–1102
Fortier ME, Joober R, Luheshi GN, Boksa P (2004) Maternal
exposure to bacterial endotoxin during pregnancy enhances
amphetamine-induced locomotion and startle responses in adult
rat offspring. J Psychiatr Res 38:335–345
Fox C, Merali Z, Harrison C (2006) Therapeutic and protective effect
of environmental enrichment against psychogenic and neuro-
genic stress. Behav Brain Res 175:1–8
Francis DD, Meaney MJ (1999) Maternal care and the development
of stress responses. Curr Opin Neurobiol 9:128–134
Francis DD, Diorio J, Plotsky PM, Meaney MJ (2002) Environmental
enrichment reverses the effects of maternal separation on stress
reactivity. J Neurosci 22:7840–7843
Freud S (1918) From the history of an infantile neurosis. Hogarth
Press, London
Fritzsche M (2001) Are cannabinoid receptor knockout mice animal
models for schizophrenia? Med Hypotheses 56:638–643
Giedd JN (2008) The teen brain: insights from neuroimaging. J
Adolesc Health 42:335–343
Giedd JN, Lalonde FM, Celano MJ, White SL, Wallace GL, Lee NR,
Lenroot RK (2009) Anatomical brain magnetic resonance
imaging of typically developing children and adolescents. J
Am Acad Child Adolesc Psychiatr 48:465–470
Goggi J, Pullar IA, Carney SL, Bradford HF (2002) Modulation of
neurotransmitter release induced by brain-derived neurotrophic
factor in rat brain striatal slices in vitro. Brain Res 941:34–42
Gottesman II, Hanson DR (2005) Human development: biological
and genetic processes. Annu Rev Psychol 56:263–286
Gotz AA, Stefanski V (2007) Psychosocial maternal stress during
pregnancy affects serum corticosterone, blood immune param-
eters and anxiety behaviour in adult male rat offspring. Physiol
Behav 90:108–115
Greaves-Lord K, Huizink AC, Oldehinkel AJ, Ormel J, Verhulst FC,
Ferdinand RF (2009) Baseline cortisol measures and
Neurotox Res (2011) 19:286–307
developmental pathways of anxiety in early adolescence. Acta
psychiatrica Scandinavica 120:178–186
Grech A, Van Os J, Jones PB, Lewis SW, Murray RM (2005)
Cannabis use and outcome of recent onset psychosis. Eur
Psychiatr 20:349–353
Green AI, Canuso CM, Brenner MJ, Wojcik JD (2003) Detection and
management of comorbidity in patients with schizophrenia.
Psychiatr Clin North Am 26:115–139
Grojean S, Schroeder H, Pourie G, Charriaut-Marlangue C, Koziel V,
Desor D, Vert P, Daval JL (2003) Histopathological alterations
and functional brain deficits after transient hypoxia in the newborn
rat pup: a long term follow-up. Neurobiol Dis 14:265–278
Guillin O, Abi-Dargham A, Laruelle M (2007) Neurobiology of
dopamine in schizophrenia. Int Rev Neurobiol 78:1–39
Hall W, Degenhardt L (2008) Cannabis use and the risk of developing
a psychotic disorder. World Psychiatr 7:68–71
Harlow HF, Harlow MK (1965) The affectional systems. In: Schrier
AM, Harlow HF, Stollnitz, F (eds) Behavior of nonhuman
primates. Academic Press, New York and London, pp 287–334
Harlow HF, Rowland GL, Griffin GA (1964) The effect of total social
deprivation on the development of monkey behavior. Psychiatr
Res Rep Am Psychiatr Assoc 19:116–135
Heim C, Nemeroff CB (2001) The role of childhood trauma in the
neurobiology of mood and anxiety disorders: preclinical and
clinical studies. Biol Psychiatr 49:1023–1039
Heim C, Newport DJ, Mletzko T, Miller AH, Nemeroff CB (2008)
The link between childhood trauma and depression: insights
from HPA axis studies in humans. Psychoneuroendocrinology
33:693–710
Iuvone L, Geloso MC, Dell’Anna E (1996) Changes in open field
behavior, spatial memory, and hippocampal parvalbumin immu-
noreactivity following enrichment in rats exposed to neonatal
anoxia. Exp Neurol 139:25–33
Johansen EB, Aase H, Meyer A, Sagvolden T (2002) Attention-
deficit/hyperactivity disorder (ADHD) behaviour explained by
dysfunctioning reinforcement and extinction processes. Behav
Brain Res 130:37–45
Johnson MH (2005) Sensitive periods in functional brain develop-
ment: problems and prospects. Dev Psychobiol 46:287–292
Kandel DB, Yamaguchi K, Chen K (1992) Stages of progression in
drug involvement from adolescence to adulthood: further
evidence for the gateway theory. J Stud Alcohol 53:447–457
Kelley AE, Schochet T, Landry CF (2004) Risk taking and novelty
seeking in adolescence: introduction to part I. Ann NY Acad Sci
1021:27–32
Kendler KS, Sheth K, Gardner CO, Prescott CA (2002) Childhood
parental loss and risk for first-onset of major depression and
alcohol dependence: the time-decay of risk and sex differences.
Psychol Med 32:1187–1194
Koenig JI, Elmer GI, Shepard PD, Lee PR, Mayo C, Joy B, Hercher
E, Brady DL (2005) Prenatal exposure to a repeated variable
stress paradigm elicits behavioral and neuroendocrinological
changes in the adult offspring: potential relevance to schizo-
phrenia. Behav Brain Res 156:251–261
Kofman O (2002) The role of prenatal stress in the etiology of
developmental behavioural disorders. Neurosci Biobehav Rev
26:457–470
Kolevzon A, Gross R, Reichenberg A (2007) Prenatal and perinatal
risk factors for autism: a review and integration of findings. Arch
Pediatr Adolesc Med 161:326–333
Kostrzewa RM, Reader TA, Descarries L (1998) Serotonin neural
adaptations to ontogenetic loss of dopamine neurons in rat brain.
J Neurochem 70:889–898
La Greca AM, Prinstein MJ, Fetter MD (2001) Adolescent peer crowd
affiliation: linkages with health-risk behaviors and close friend-
ships. J Pediatr Psychol 26:131–143
303
Lai S, Lai H, Page JB, McCoy CB (2000) The association between
cigarette smoking and drug abuse in the United States. J Addict
Dis 19:11–24
Laviola G, Adriani W, Terranova ML, Gerra G (1999) Psychobio-
logical risk factors for vulnerability to psychostimulants in
human adolescents and animal models. Neurosci Biobehav Rev
23:993–1010
Laviola G, Macrı̀ S, Morley-Fletcher S, Adriani W (2003) Risk-taking
behavior in adolescent mice: psychobiological determinants and
early epigenetic influence. Neurosci Biobehav Rev 27:19–31
Laviola G, Adriani W, Rea M, Aloe L, Alleva E (2004a) Social
withdrawal, neophobia, and stereotyped behavior in developing
rats exposed to neonatal asphyxia. Psychopharmacology (Berl)
175:196–205
Laviola G, Rea M, Morley-Fletcher S, Di Carlo S, Bacosi A, De
Simone R, Bertini M, Pacifici R (2004b) Beneficial effects of
enriched environment on adolescent rats from stressed pregnan-
cies. Eur J Neurosci 20:1655–1664
Laviola G, Hannan AJ, Macrı̀ S, Solinas M, Jaber M (2008) Effects of
enriched environment on animal models of neurodegenerative
diseases and psychiatric disorders. Neurobiol Dis 31:159–168
Laviola G, Ognibene E, Romano E, Adriani W, Keller F (2009) Gene-
environment interaction during early development in the
heterozygous reeler mouse: clues for modelling of major
neurobehavioral syndromes. Neurosci Biobehav Rev 33:560–
572
Law AJ, Pei Q, Walker M, Gordon-Andrews H, Weickert CS, Feldon
J, Pryce CR, Harrison PJ (2009) Early parental deprivation in the
marmoset monkey produces long-term changes in hippocampal
expression of genes involved in synaptic plasticity and impli-
cated in mood disorder. Neuropsychopharmacology 34:1381–
1394
Lazinski MJ, Shea AK, Steiner M (2008) Effects of maternal prenatal
stress on offspring development: a commentary. Arch Womens
Ment Health 11:363–375
Lee JH, Kim HJ, Kim JG, Ryu V, Kim BT, Kang DW, Jahng JW
(2007a) Depressive behaviors and decreased expression of
serotonin reuptake transporter in rats that experienced neonatal
maternal separation. Neurosci Res 58:32–39
Lee PR, Brady DL, Shapiro RA, Dorsa DM, Koenig JI (2007b)
Prenatal stress generates deficits in rat social behavior: reversal
by oxytocin. Brain Res 1156:152–167
Lehmann J, Pryce CR, Jongen-Relo AL, Stohr T, Pothuizen HH,
Feldon J (2002) Comparison of maternal separation and early
handling in terms of their neurobehavioral effects in aged rats.
Neurobiol Aging 23:457–466
Lemaire V, Koehl M, Le Moal M, Abrous DN (2000) Prenatal stress
produces learning deficits associated with an inhibition of
neurogenesis in the hippocampus. Proc Natl Acad Sci USA
97:11032–11037
Lemaire V, Lamarque S, Le Moal M, Piazza PV, Abrous DN (2006)
Postnatal stimulation of the pups counteracts prenatal stress-
induced deficits in hippocampal neurogenesis. Biol Psychiatr
59:786–792
Leo D, Adriani W, Cavaliere C, Cirillo G, Marco EM, Romano E, di
Porzio U, Papa M, Perrone-Capano C, Laviola G (2009)
Methylphenidate to adolescent rats drives enduring changes of
accumbal Htr7 expression: implications for impulsive behavior
and neuronal morphology. Genes Brain Behav 8:356–368
Levine S (1957) Infantile experience and resistance to physiological
stress. Science 126:405
Levine S (2000) Influence of psychological variables on the activity
of the hypothalamic-pituitary-adrenal axis. Eur J Pharmacol
405:149–160
Levine S (2005) Developmental determinants of sensitivity and
resistance to stress. Psychoneuroendocrinology 30:939–946
123
304
Levine S, Alpert M, Lewis GW (1957) Infantile experience and the
maturation of the pituitary adrenal axis. Science 126:1347
Levine S, Huchton DM, Wiener SG, Rosenfeld P (1991) Time course
of the effect of maternal deprivation on the hypothalamic-
pituitary-adrenal axis in the infant rat. Dev Psychobiol 24:547–
558
Leweke FM, Koethe D (2008) Cannabis and psychiatric disorders: it
is not only addiction. Addict Biol 13:264–275
Leweke FM, Giuffrida A, Wurster U, Emrich HM, Piomelli D (1999)
Elevated endogenous cannabinoids in schizophrenia. Neurore-
port 10:1665–1669
Liu D, Diorio J, Tannenbaum B, Caldji C, Francis D, Freedman A,
Sharma S, Pearson D, Plotsky PM, Meaney MJ (1997) Maternal
care, hippocampal glucocorticoid receptors, and hypothalamic-
pituitary-adrenal responses to stress. Science 277:1659–1662
Llorente E, Brito ML, Machado P, Gonzalez MC (2002) Effect of
prenatal stress on the hormonal response to acute and chronic
stress and on immune parameters in the offspring. J Physiol
Biochem 58:143–149
Llorente R, Arranz L, Marco EM, Moreno E, Puerto M, Guaza C, De
la FM, Viveros MP (2007) Early maternal deprivation and
neonatal single administration with a cannabinoid agonist induce
long-term sex-dependent psychoimmunoendocrine effects in
adolescent rats. Psychoneuroendocrinology 32:636–650
Llorente R, Llorente-Berzal A, Petrosino S, Marco EM, Guaza C,
Prada C, Lopez-Gallardo M, Di Marzo V, Viveros MP (2008)
Gender-dependent cellular and biochemical effects of maternal
deprivation on the hippocampus of neonatal rats: a possible role
for the endocannabinoid system. Dev Neurobiol 68:1334–1347
Lockwood CA, Menter CG, Moggi-Cecchi J, Keyser AW (2007)
Extended male growth in a fossil hominin species. Science
318:1443–1446
Lorenz K (1937/1957) The conception of instinctive behavior. In:
Schiller CH (ed) Instinctive behavior. International Universities
Press, New York, pp 129–175
Lowry CA, Hale MW, Evans AK, Heerkens J, Staub DR, Gasser PJ,
Shekhar A (2008) Serotonergic systems, anxiety, and affective
disorder: focus on the dorsomedial part of the dorsal raphe
nucleus. Ann N Y Acad Sci 1148:86–94
Maccari S, Morley-Fletcher S (2007) Effects of prenatal restraint
stress on the hypothalamus-pituitary-adrenal axis and related
behavioural and neurobiological alterations. Psychoneuroendo-
crinology 32 Suppl 1:S10–S15
Maccari S, Darnaudery M, Morley-Fletcher S, Zuena AR, Cinque C,
Van Reeth O (2003) Prenatal stress and long-term consequences:
implications of glucocorticoid hormones. Neurosci Biobehav
Rev 27:119–127
Mackowiak M, Chocyk A, Markowicz-Kula K, Wedzony K (2004)
Neurogenesis in the adult brain. Pol J Pharmacol 56:673–687
Macrı̀ S, Chiarotti F, Wurbel H (2008) Maternal separation and
maternal care act independently on the development of HPA
responses in male rats. Behav Brain Res 191:227–234
Macrı̀ S, Laviola G (2004) Single episode of maternal deprivation and
adult depressive profile in mice: interaction with cannabinoid
exposure during adolescence. Behav Brain Res 154:231–238
Macrı̀ S, Wurbel H (2006) Developmental plasticity of HPA and fear
responses in rats: a critical review of the maternal mediation
hypothesis. Horm Behav 50:667–680
Macrı̀ S, Mason GJ, Wurbel H (2004) Dissociation in the effects of
neonatal maternal separations on maternal care and the off-
spring’s HPA and fear responses in rats. Eur J Neurosci
20:1017–1024
Macrı̀ S, Pasquali P, Bonsignore LT, Pieretti S, Cirulli F, Chiarotti F,
Laviola G (2007a) Moderate neonatal stress decreases within-
group variation in behavioral, immune and HPA responses in
adult mice. PLoS One 2:e1015
123
Neurotox Res (2011) 19:286–307
Macrı̀ S, Spinelli S, Adriani W, Dee Higley J, Laviola G (2007b)
Early adversity and alcohol availability persistently modify
serotonin and hypothalamic-pituitary-adrenal-axis metabolism
and related behavior: what experimental research on rodents and
primates can tell us. Neurosci Biobehav Rev 31:172–180
Macrı̀ S, Granstrem O, Shumilina M, Antunes Gomes dos Santos FJ,
Berry A, Saso L, Laviola G (2009) Resilience and vulnerability
are dose-dependently related to neonatal stressors in mice. Horm
Behav 56:391–398
Maneru C, Junque C, Botet F, Tallada M, Guardia J (2001)
Neuropsychological long-term sequelae of perinatal asphyxia.
Brain Inj 15:1029–1039
Marchetti B, Morale MC, Testa N, Tirolo C, Caniglia S, Amor S,
Dijkstra CD, Barden N (2001) Stress, the immune system and
vulnerability to degenerative disorders of the central nervous
system in transgenic mice expressing glucocorticoid receptor
antisense RNA. Brain Res Brain Res Rev 37:259–272
Marco EM, Granstrem O, Moreno E, Llorente R, Adriani W, Laviola
G, Viveros MP (2007) Subchronic nicotine exposure in adoles-
cence induces long-term effects on hippocampal and striatal
cannabinoid-CB1 and mu-opioid receptors in rats. Eur J
Pharmacol 557:37–43
Marco EM, Adriani W, Llorente R, Laviola G, Viveros MP (2009)
Detrimental psychophysiological effects of early maternal
deprivation in adolescent and adult rodents: altered responses
to cannabinoid exposure. Neurosci Biobehav Rev 33:498–507
Matthews K, Robbins TW (2003) Early experience as a determinant
of adult behavioural responses to reward: the effects of repeated
maternal separation in the rat. Neurosci Biobehav Rev 27:45–55
McBride WJ, Bell RL, Rodd ZA, Strother WN, Murphy JM (2005)
Adolescent alcohol drinking and its long-range consequences.
Studies with animal models. Recent Dev Alcohol 17:123–142
Meaney MJ (2001) Maternal care, gene expression, and the
transmission of individual differences in stress reactivity across
generations. Annu Rev Neurosci 24:1161–1192
Meaney MJ, Diorio J, Francis D, Weaver S, Yau J, Chapman K, Seckl
JR (2000) Postnatal handling increases the expression of cAMP-
inducible transcription factors in the rat hippocampus: the effects
of thyroid hormones and serotonin. J Neurosci 20:3926–3935
Meijer A (1985) Child psychiatric sequelae of maternal war stress.
Acta Psychiatr Scand 72:505–511
Merlot E, Couret D, Otten W (2008) Prenatal stress, fetal imprinting
and immunity. Brain Behav Immun 22:42–51
Meyer U, Feldon J, Fatemi SH (2009) In-vivo rodent models for the
experimental investigation of prenatal immune activation effects
in neurodevelopmental brain disorders. Neurosci Biobehav Rev
33:1061–1079
Michel GF, Tyler AN (2005) Critical period: a history of the
transition from questions of when, to what, to how. Dev
Psychobiol 46:156–162
Mill J, Petronis A (2008) Pre- and peri-natal environmental risks for
attention-deficit hyperactivity disorder (ADHD): the potential
role of epigenetic processes in mediating susceptibility. J Child
Psychol Psychiatry 49:1020–1030
Mittal VA, Ellman LM, Cannon TD (2008) Gene-environment
interaction and covariation in schizophrenia: the role of obstetric
complications. Schizophr Bull 34:1083–1094
Moffett MC, Vicentic A, Kozel M, Plotsky P, Francis DD, Kuhar MJ
(2007) Maternal separation alters drug intake patterns in
adulthood in rats. Biochem Pharmacol 73:321–330
Montoya AG, Sorrentino R, Lukas SE, Price BH (2002) Long-term
neuropsychiatric consequences of ‘‘ecstasy’’ (MDMA): a review.
Harv Rev Psychiatr 10:212–220
Morgan C, Fisher H (2007) Environment and schizophrenia:
environmental factors in schizophrenia: childhood trauma—a
critical review. Schizophr Bull 33:3–10
Neurotox Res (2011) 19:286–307
Morley-Fletcher S, Darnaudery M, Koehl M, Casolini P, Van Reeth
O, Maccari S (2003a) Prenatal stress in rats predicts immobility
behavior in the forced swim test. Effects of a chronic treatment
with tianeptine. Brain Res 989:246–251
Morley-Fletcher S, Rea M, Maccari S, Laviola G (2003b) Environ-
mental enrichment during adolescence reverses the effects of
prenatal stress on play behaviour and HPA axis reactivity in rats.
Eur J Neurosci 18:3367–3374
Moy SS, Nadler JJ (2008) Advances in behavioral genetics: mouse
models of autism. Mol Psychiatr 13:4–26
Muller N, Riedel M, Gruber R, Ackenheil M, Schwarz MJ (2000) The
immune system and schizophrenia. An integrative view. Ann NY
Acad Sci 917:456–467
Nemeroff CB, Owens MJ (2009) The role of serotonin in the
pathophysiology of depression: as important as ever. Clin Chem
55:1578–1579
Nithianantharajah J, Hannan AJ (2006) Enriched environments,
experience-dependent plasticity and disorders of the nervous
system. Nat Rev Neurosci 7:697–709
Nocjar C, Roth BL, Pehek EA (2002) Localization of 5-HT(2A)
receptors on dopamine cells in subnuclei of the midbrain A10
cell group. Neuroscience 111:163–176
Nowakowski RS, Hayes NL (1999) CNS development: an overview.
Dev Psychopathol 11:395–417
O’Donnell K, O’Connor TG, Glover V (2009) Prenatal stress and
neurodevelopment of the child: focus on the HPA axis and role
of the placenta. Dev Neurosci 31:285–292
O’Shea M, Singh ME, McGregor IS, Mallet PE (2004) Chronic
cannabinoid exposure produces lasting memory impairment and
increased anxiety in adolescent but not adult rats. J Psychophar-
macol 18:502–508
O’Tuathaigh CM, Babovic D, O’Meara G, Clifford JJ, Croke DT,
Waddington JL (2007) Susceptibility genes for schizophrenia:
characterisation of mutant mouse models at the level of
phenotypic behaviour. Neurosci Biobehav Rev 31:60–78
Painter MJ (1995) Animal models of perinatal asphyxia: contributions,
contradictions, clinical relevance. Semin Pediatr Neurol 2:37–56
Patterson PH (2007) Neuroscience. Maternal effects on schizophrenia
risk. Science 318:576–577
Patterson PH (2009) Immune involvement in schizophrenia and
autism: etiology, pathology and animal models. Behav Brain Res
204:313–321
Penner JD, Brown AS (2007) Prenatal infectious and nutritional
factors and risk of adult schizophrenia. Expert Rev Neurother
7:797–805
Pereira LO, Strapasson AC, Nabinger PM, Achaval M, Netto CA
(2008) Early enriched housing results in partial recovery of
memory deficits in female, but not in male, rats after neonatal
hypoxia-ischemia. Brain Res 1218:257–266
Pereira LO, Nabinger PM, Strapasson AC, Nardin P, Goncalves CA,
Siqueira IR, Netto CA (2009) Long-term effects of environmen-
tal stimulation following hypoxia-ischemia on the oxidative state
and BDNF levels in rat hippocampus and frontal cortex. Brain
Res 1247:188–195
Phillips LJ, McGorry PD, Garner B, Thompson KN, Pantelis C, Wood
SJ, Berger G (2006) Stress, the hippocampus and the
hypothalamic-pituitary-adrenal axis: implications for the devel-
opment of psychotic disorders. Aust NZ J Psychiatr 40:725–741
Pickering C, Gustafsson L, Cebere A, Nylander I, Liljequist S (2006)
Repeated maternal separation of male Wistar rats alters gluta-
mate receptor expression in the hippocampus but not the
prefrontal cortex. Brain Res 1099:101–108
Piper BJ (2007) A developmental comparison of the neurobehavioral
effects of ecstasy (MDMA). Neurotoxicol Teratol 29:288–300
Plotsky PM, Meaney MJ (1993) Early, postnatal experience alters
hypothalamic corticotropin-releasing factor (CRF) mRNA,
305
median eminence CRF content and stress-induced release in
adult rats. Brain Res Mol Brain Res 18:195–200
Primus RJ, Kellogg CK (1989) Pubertal-related changes influence the
development of environment-related social interaction in the
male rat. Dev Psychobiol 22:633–643
Pryce CR, Feldon J (2003) Long-term neurobehavioural impact of the
postnatal environment in rats: manipulations, effects and medi-
ating mechanisms. Neurosci Biobehav Rev 27:57–71
Raju TN (1992) Some animal models for the study of perinatal
asphyxia. Biol Neonate 62:202–214
Rasmuson S, Olsson T, Henriksson BG, Kelly PA, Holmes MC, Seckl
JR, Mohammed AH (1998) Environmental enrichment selec-
tively increases 5-HT1A receptor mRNA expression and binding
in the rat hippocampus. Brain Res Mol Brain Res 53:285–290
Rice D, Barone S Jr (2000) Critical periods of vulnerability for the
developing nervous system: evidence from humans and animal
models. Environ Health Perspect 108 (Suppl 3):511–533
Rice F, Jones I, Thapar A (2007) The impact of gestational stress and
prenatal growth on emotional problems in offspring: a review.
Acta Psychiatr Scand 115:171–183
Rice F, Harold GT, Boivin J, van den Bree M, Hay DF, Thapar A
(2009) The links between prenatal stress and offspring develop-
ment and psychopathology: disentangling environmental and
inherited influences. Psychol Med 1–11
Roceri M, Hendriks W, Racagni G, Ellenbroek BA, Riva MA (2002)
Early maternal deprivation reduces the expression of BDNF and
NMDA receptor subunits in rat hippocampus. Mol Psychiatr
7:609–616
Rosenzweig MR, Bennett EL (1996) Psychobiology of plasticity:
effects of training and experience on brain and behavior. Behav
Brain Res 78:57–65
Rosenzweig MR, Bennett EL, Hebert M, Morimoto H (1978) Social
grouping cannot account for cerebral effects of enriched
environments. Brain Res 153:563–576
Rozeboom AM, Akil H, Seasholtz AF (2007) Mineralocorticoid
receptor overexpression in forebrain decreases anxiety-like
behavior and alters the stress response in mice. Proc Natl Acad
Sci USA 104:4688–4693
Rubino T, Parolaro D (2008) Long lasting consequences of cannabis
exposure in adolescence. Mol Cell Endocrinol 286:S108–S113
Ruedi-Bettschen D, Pedersen EM, Feldon J, Pryce CR (2005) Early
deprivation under specific conditions leads to reduced interest in
reward in adulthood in Wistar rats. Behav Brain Res 156:297–
310
Rufini O, Valanzano A, Calamandrei G (2001) Animal models of
hypoxic-ischemic encephalopathy. Ann Ist Super Sanita 37:561–
566
Sarkar NN (2008) The impact of intimate partner violence on
women’s reproductive health and pregnancy outcome. J Obstet
Gynaecol 28:266–271
Sawamura N, Sawa A (2006) Disrupted-in-schizophrenia-1 (DISC1):
a key susceptibility factor for major mental illnesses. Ann NY
Acad Sci 1086:126–133
Scheepens A, Wassink G, Blanco CE (2003) The effect of a global
birth asphyxia on the ontogeny of BDNF and NGF protein
expression in the juvenile brain. Brain Res Dev Brain Res
140:215–221
Schepis TS, Adinoff B, Rao U (2008) Neurobiological processes in
adolescent addictive disorders. Am J Addict 17:6–23
Schiepers OJ, Wichers MC, Maes M (2005) Cytokines and major
depression. Prog Neuropsychopharmacol Biol Psychiatr 29:201–
217
Schmidt M, Enthoven L, van Woezik JH, Levine S, de Kloet ER,
Oitzl MS (2004) The dynamics of the hypothalamic-pituitary-
adrenal axis during maternal deprivation. J Neuroendocrinol
16:52–57
123
306
Schneider M (2008) Puberty as a highly vulnerable developmental
period for the consequences of cannabis exposure. Addict Biol
13:253–263
Schneider M, Koch M (2003) Chronic pubertal, but not adult chronic
cannabinoid treatment impairs sensorimotor gating, recognition
memory, and the performance in a progressive ratio task in adult
rats. Neuropsychopharmacology 28:1760–1769
Schneirla TC, Rosenblatt JS (1963) ‘‘Critical Periods’’ in the
Development of Behavior. Science 139:1110–1115
Schwarz MJ, Chiang S, Muller N, Ackenheil M (2001) T-helper-1
and T-helper-2 responses in psychiatric disorders. Brain Behav
Immun 15:340–370
Scott JP (1962) Critical periods in behavioral development. Science
138:949–958
Scott J, McNeill Y, Cavanagh J, Cannon M, Murray R (2006)
Exposure to obstetric complications and subsequent develop-
ment of bipolar disorder: systematic review. Br J Psychiatr
189:3–11
Shea A, Walsh C, Macmillan H, Steiner M (2005) Child maltreatment
and HPA axis dysregulation: relationship to major depressive
disorder and post traumatic stress disorder in females. Psycho-
neuroendocrinology 30:162–178
Shi L, Fatemi SH, Sidwell RW, Patterson PH (2003) Maternal
influenza infection causes marked behavioral and pharmacolog-
ical changes in the offspring. J Neurosci 23:297–302
Siqueira LM, Brook JS (2003) Tobacco use as a predictor of illicit
drug use and drug-related problems in Colombian youth. J
Adolesc Health 32:50–57
Skosnik PD, Spatz-Glenn L, Park S (2001) Cannabis use is associated
with schizotypy and attentional disinhibition. Schizophr Res
48:83–92
Slotkin TA (2002) Nicotine and the adolescent brain: insights from an
animal model. Neurotoxicol Teratol 24:369–384
Slotkin TA, MacKillop EA, Rudder CL, Ryde IT, Tate CA, Seidler FJ
(2007) Permanent, sex-selective effects of prenatal or adolescent
nicotine exposure, separately or sequentially, in rat brain
regions: indices of cholinergic and serotonergic synaptic func-
tion, cell signaling, and neural cell number and size at 6 months
of age. Neuropsychopharmacology 32:1082–1097
Smotherman WP, Bell RW (1980) Maternal mediation of early
experience. In: Smotherman WP, Bell RL (eds) Maternal
influences and early behavior. Spectrum, New York, pp 201–210
Sondeijker FE, Ferdinand RF, Oldehinkel AJ, Tiemeier H, Ormel J,
Verhulst FC (2008) HPA-axis activity as a predictor of future
disruptive behaviors in young adolescents. Psychophysiology
45:398–404
Spear LP (2000) The adolescent brain and age-related behavioral
manifestations. Neurosci Biobehav Rev 24:417–463
Stein DJ (2008) Depression, anhedonia, and psychomotor symp-
toms: the role of dopaminergic neurocircuitry. CNS Spectr
13:561–565
Stern JM, Johnson SK (1990) Ventral somatosensory determinants of
nursing behavior in Norway rats. I. Effects of variations in the
quality and quantity of pup stimuli. Physiol Behav 47:993–1011
Strous RD, Shoenfeld Y (2006) Schizophrenia, autoimmunity and
immune system dysregulation: a comprehensive model updated
and revisited. J Autoimmun 27:71–80
Suárez J, Llorente R, Romero-Zerbo SY, Mateos B, Bermudez-Silva
FJ, de Fonseca FR, Viveros MP (2009) Early maternal depri-
vation induces gender-dependent changes on the expression of
hippocampal CB(1) and CB(2) cannabinoid receptors of neonatal
rats. Hippocampus 19:623–632
Suchecki D, Tufik S (1997) Long-term effects of maternal deprivation
on the corticosterone response to stress in rats. Am J Physiol
273:R1332–R1338
123
Neurotox Res (2011) 19:286–307
Susman EJ (2006) Psychobiology of persistent antisocial behavior:
stress, early vulnerabilities and the attenuation hypothesis.
Neurosci Biobehav Rev 30:376–389
Talge NM, Neal C, Glover V (2007) Antenatal maternal stress and
long-term effects on child neurodevelopment: how and why? J
Child Psychol Psychiatr 48:245–261
Teicher MH, Tomoda A, Andersen SL (2006) Neurobiological
consequences of early stress and childhood maltreatment: are
results from human and animal studies comparable? Ann NY
Acad Sci 1071:313–323
Terranova ML, Laviola G, de Acetis L, Alleva E (1998) A description
of the ontogeny of mouse agonistic behavior. J Comp Psychol
112:3–12
Tohmi M, Tsuda N, Watanabe Y, Kakita A, Nawa H (2004) Perinatal
inflammatory cytokine challenge results in distinct neurobehav-
ioral alterations in rats: implication in psychiatric disorders of
developmental origin. Neurosci Res 50:67–75
Tuchscherer M, Kanitz E, Otten W, Tuchscherer A (2002) Effects of
prenatal stress on cellular and humoral immune responses in
neonatal pigs. Vet Immunol Immunopathol 86:195–203
Ujike H, Morita Y (2004) New perspectives in the studies on
endocannabinoid and cannabis: cannabinoid receptors and
schizophrenia. J Pharmacol Sci 96:376–381
Van den Hove DL, Steinbusch HW, Scheepens A, Van de Berg WD,
Kooiman LA, Boosten BJ, Prickaerts J, Blanco CE (2006)
Prenatal stress and neonatal rat brain development. Neuroscience
137:145–155
van Handel M, Swaab H, de Vries LS, Jongmans MJ (2007) Long-
term cognitive and behavioral consequences of neonatal enceph-
alopathy following perinatal asphyxia: a review. Eur J Pediatr
166:645–654
van Oers HJ, de Kloet ER, Levine S (1997) Persistent, but
paradoxical, effects on HPA regulation of infants maternally
deprived at different ages. Stress 1:249–262
van Praag H, Kempermann G, Gage FH (2000) Neural consequences
of environmental enrichment. Nat Rev Neurosci 1:191–198
Vanderschuren LJ, Niesink RJ, Van Ree JM (1997) The neurobiology
of social play behavior in rats. Neurosci Biobehav Rev 21:309–
326
Vidal J, Bie J, Granneman RA, Wallinga AE, Koolhaas JM, Buwalda
B (2007) Social stress during adolescence in Wistar rats induces
social anxiety in adulthood without affecting brain monoamin-
ergic content and activity. Physiol behav 92:824–830
Wakuda T, Matsuzaki H, Suzuki K, Iwata Y, Shinmura C, Suda S,
Iwata K, Yamamoto S, Sugihara G, Tsuchiya KJ, Ueki T,
Nakamura K, Nakahara D, Takei N, Mori N (2008) Perinatal
asphyxia reduces dentate granule cells and exacerbates meth-
amphetamine-induced hyperlocomotion in adulthood. PLoS One
3:e3648
Ward IL, Stehm KE (1991) Prenatal stress feminizes juvenile play
patterns in male rats. Physiol Behav 50:601–605
Watanabe Y, Hashimoto S, Kakita A, Takahashi H, Ko J, Mizuno M,
Someya T, Patterson PH, Nawa H (2004) Neonatal impact of
leukemia inhibitory factor on neurobehavioral development in
rats. Neurosci Res 48:345–353
Weaver IC, Cervoni N, Champagne FA, D’Alessio AC, Sharma S,
Seckl JR, Dymov S, Szyf M, Meaney MJ (2004) Epigenetic
programming by maternal behavior. Nat Neurosci 7:847–854
Weininger O (1954) Physiological damage under emotional stress as a
function of early experience. Science 119:285–286
Weininger O, Mc CW, Arima RK (1954) Gentling and weight gain in
the albino rat. Can J Psychol 8:147–151
Weinstock M (2002) Can the behaviour abnormalities induced by
gestational stress in rats be prevented or reversed? Stress 5:167–
176
Neurotox Res (2011) 19:286–307
Weinstock M (2007) Gender differences in the effects of prenatal
stress on brain development and behaviour. Neurochem Res
32:1730–1740
Weinstock M (2008) The long-term behavioural consequences of
prenatal stress. Neurosci Biobehav Rev 32:1073–1086
Wetzels JJ, Kremers SP, Vitoria PD, de Vries H (2003) The alcohol-
tobacco relationship: a prospective study among adolescents in
six European countries. Addiction 98:1755–1763
Yolken RH, Karlsson H, Yee F, Johnston-Wilson NL, Torrey EF
(2000) Endogenous retroviruses and schizophrenia. Brain Res
Brain Res Rev 31:193–199
Zimmerberg B, Shartrand AM (1992) Temperature-dependent effects
of maternal separation on growth, activity, and amphetamine
sensitivity in the rat. Dev Psychobiol 25:213–226
307
Zuckerman L, Rehavi M, Nachman R, Weiner I (2003) Immune
activation during pregnancy in rats leads to a postpubertal
emergence of disrupted latent inhibition, dopaminergic hyper-
function, and altered limbic morphology in the offspring: a novel
neurodevelopmental model of schizophrenia. Neuropsychophar-
macology 28:1778–1789
Zuena AR, Mairesse J, Casolini P, Cinque C, Alema GS, Morley-
Fletcher S, Chiodi V, Spagnoli LG, Gradini R, Catalani A,
Nicoletti F, Maccari S (2008) Prenatal restraint stress generates
two distinct behavioral and neurochemical profiles in male and
female rats. PLoS One 3:e2170
123