ZOOLOGICAL RESEARCH Consequences of early adverse rearing experience (EARE) on development: insights from non-human primate studies Bo Zhang1,2,3,* 1 Yunnan Key Laboratory of Primate Biomedical Research, Kunming Yunnan 650500, China 2 Institute of Primate Translational Medicine, Kunming University of Science and Technology, Kunming Yunnan 650500, China 3 National Institute of Health, Bethesda, Maryland, USA ABSTRACT individual's social relationship throughout life is influenced by the initial attachment with the mother (Bowlby, 1969). Early rearing experiences are important in one’s Attachment theory is a psychological, evolutionary and whole life, whereas early adverse rearing experience ethological theory concerning relationships among humans. (EARE) is usually related to various physical and Within the theory, attachment means an affectional bond or tie mental disorders in later life. Although there were between an individual and an attachment figure (usually a many studies on human and animals, regarding the caregiver). The core is that a child needs to build relationship effect of EARE on brain development, neuroendocrine with at least one primary caregiver to develop normal social and systems, as well as the consequential mental disorders emotional behaviors. In many orphans, the lack of normal and behavioral abnormalities, the underlying attachment to parents would cause behavioral and physical mechanisms remain unclear. Due to the close problems in childhood and possibly continuing throughout adult genetic relationship and similarity in social life (McEwen, 2003). Adults with adverse experience were more organizations with humans, non-human primate vulnerable to physical, psychosocial and mental disorders (NHP) studies were performed for over 60 years. (Maughan & McCarthy, 1997; 1Pirkola et al., 2005). In human, Various EARE models were developed to disrupt the early adverse rearing experience (EARE) usually refers to child early normal interactions between infants and abuse, which is a worldwide problem and is defined as neglect mothers or peers. Those studies provided important or physical, sexual or emotional mistreatment or abuse of insights of EARE induced effects on the physiological children (Newton & Vandeven, 2009, 2010). Although human and behavioral systems of NHPs across life span, based studies revealed compelling associations between EARE such as social behaviors (including disturbance and psychological outcomes, both retrospective and behavior, social deficiency, sexual behavior, etc), prospective studies showed their limits, e.g., inaccurate self- learning and memory ability, brain structural and report due to biased or even false memory, failure in controlling functional developments (including influences on accompanying environmental and genetic factors. Therefore, neurons and glia cells, neuroendocrine systems, e.g., the long-term effects of EARE on subjects were usually not hypothalamic-pituitary-adrenal (HPA) axis, etc). In the direct consequences, but were inevitably intervened or this review, the effects of EARE and the underlying masked by uncontrollable factors. However, experimental epigenetic mechanisms were comprehensively animals can be raised in laboratory environments, therefore summarized and the possibility of rehabilitation was allow researchers to carry out randomized prospective discussed. longitudinal studies, e.g., rigorously control or systematically manipulate early experiences throughout the entire period of Keywords: Early adverse rearing experience; Non- investigation. human primates INTRODUCTION Received: 01 December 2016; Accepted: 30 December 2016 Foundation items: This research was supported by Hainan special One of factors affecting life-long health of humans is the stability fund project for science and technology (KJHZ2015-20) of early childhood, especially children’s relationship with their *Corresponding author, E-mail: [email protected] mothers. John Bowlby's attachment theory suggests that DOI:10.13918/j.issn.2095-8137.2017.002 Science Press Zoological Research 38(1): 7-35, 2017 7 Rodents are easy to manipulate genetically, and the related learning and memory ability, brain structural and functional studies indicate EARE as a developmental risk factor with development (e.g., development of neurons and glia cells, profound, long-term effects on later life (Meaney, 2001; Pryce et neuroendocrine dysregulation, etc). In this review, the previous al., 2005b; Sánchez et al., 2001). Whereas the high similarities findings on EARE were systematically summarized, and the of NHPs with humans make it irreplaceable in investigating the underlying epigenetic mechanisms and the potential methods of effects of EARE on physiological and behavioral development, rehabilitation were thoroughly discussed. e.g., NHPs and chimpanzees in specific, share over 90% and 98.8% genomes with human beings, respectively (Lovejoy, EARE MODELS IN NON-HUMAN PRIMATES 1981). High similarities were found in both biological (Azmitia & Gannon, 1986; Uylings & van Eden, 1991) and socio- Controlled rearing conditions in standard laboratory settings are ecological aspects, e.g., social organizations and clear designed to simulate natural environments. The infants are dominance hierarchies (Bailey & Aunger, 1990; DeVore, 1990; reared by their mothers and live in a group consisted of other Wright, 1990). The phenomenon that in NHPs, 2%-10% of infants, juveniles and adults, allowing infants to be exposed to infants were physically abused or neglected by their mothers complex social interactions. In abnormal rearing conditions, the in group-living conditions, allow the possible screening of mother deprivation method is applied. The newborn is taken natural child abuse models (Maestripieri & Carroll, 1998; away from their mothers at birth and is reared in incubators with Maestripieri et al., 1997). Moreover, like humans NHPs has regular medical attention and laboratory nursery. A period of prolonged postnatal period of maturation during which time (usually 1-month) later, when able to feed themselves, mother–infant relationship and neural system development infants are moved to other rearing conditions depending on can be influenced by environment and early life experience aims of research, e.g., be reared alone in social isolation (Levine & Wiener, 1988; Suomi, 2005). condition, with nursery/peers of the same age in nursery/peer Harlow (Harlow & Harlow, 1965) introduced the concept of rearing condition, with a surrogate in surrogate mother/foster affectional systems to characterize the relationships in the rearing condition, etc. They could also be separated from social groups of primates, and five distinct affectional systems mothers at later time for once (temporary maternal separations) were described, including the infant-mother affectional system, or several times (repetitive maternal separations); or even the maternal affectional system, the age-mate/peer affectional though staying with their mother all the time, but still suffer from system, the heterosexual affectional system and the paternal EARE (maternal neglect). affectional system. The infant-mother and the maternal affectional systems in Harlow’s affectional systems are similar Social isolation to Bowlby’s concept of mother-infant attachment theory in Social isolation (including total and partial social isolation) is humans. In normal living group, most monkey infants virtually initially described in early 1960s by Harlow and his colleagues, spend all of their initial days or weeks of life clinging with their and has been used ever since to raise monkeys in simulating biological mothers, ventral to ventral, during which, specific and social behavior deficits in humans (Table 1). In total isolation, strong attachment bonds are built. When about 2-month old, the infant is reared in a cage alone without any auditory, visual, infants begin to explore the physical and social environment, olfactory and tactile contact with conspecifics, including mothers, spending increasing amount of time participating social peers and other monkeys (Baysinger et al., 1972; Harlow & interactions, especially playing with peers. From 6-month of age Harlow, 1962; Harlow et al., 1964, 1965). In partial social until puberty, playing with peers becomes the major social isolation, although infants are separately caged from their activity (Hinde & Spencer-Booth, 1967; Suomi, 1997, 2005). In mothers, peers, and social groups, they have auditory, visual, fact, the infant and juvenile monkeys always maintain a close and olfactory but not tactile contact with their conspecifics social relationship with their mothers, while the mother plays the (Cross & Harlow, 1965; Mason & Sponholz, 1963; Struble & role of protector especially under stressful situations, and Riesen, 1978; Suomi et al., 1971). These early studies by mentor in teaching developing appropriate social behaviors. Harlow and his colleagues, especially their extreme Accordingly, the studies regarding EARE usually involve manipulations, including total isolation, "pit of despair" and "rape disruption of the normal infant-mother relationships, by maternal rack" devices, were controversial and were most likely deprivation of newborns, maternal separation or induced stress forbidden to perform due to ethical issues. In 1950s, many on older infants and juvenile monkeys. Although some researchers assumed that the only necessity of mother was epidemiological studies in humans suggest possible direct supplying food to infants, whereas excessive intimacy relationships between EARE and abnormal behaviors in later between mother and infant would hinder the growth of infant, life, no solid evidence was raised to prove the precise impact of or even induce over dependence in adulthood. Harlow childhood adversities on psychiatric disorders (Benjet, 2010; disagreed with the viewpoints; performed a series of Bick & Nelson, 2016; Gershon et al., 2013; Kessler et al., 1997; isolation studies on primates to prove that to acquire Kessler & Wang, 2008; Klein et al., 2013; Sheridan et al., 2010). necessary social skills, to obtain both physically and The over 60 years NHP studies shed lights on the psychologically healthy development, infants need mothers’ understanding of the influences of EARE on physiological and affection, as well as normal social interaction and emotional behavioral development, including social behaviors (e.g., relationship with peers. However, their intention was to disturbance behavior, social deficiency, sexual behavior, etc), prove the essential role of mother's love to infants, in the 8 www.zoores.ac.cn Table 1 Early adverse rearing experience (EARE) methods Paradigms Description References Social Total Infants are reared in a cage alone at birth, no any auditory, visual, Baysinger et al., 1972; Harlow et al., 1965; Isolation olfactory and tactile contact with conspecifics is allowed Harlow & Harlow, 1962; Harlow et al., 1964 Partial Infants are separately caged at birth, reared with auditory, visual, and Cross & Harlow, 1965; Mason & Sponholz, olfactory, but not tactile contact with conspecifics 1963; Struble & Riesen, 1978; Suomi et al., 1971 Maternal Permanent Peer-rearing Continuous Infants are reared by pairs throughout Chamove et al., 1973; Erwin et al., 1973; separation development Sackett, 1967; Worlein & Sackett, 1997 Intermittent Peers are allowed to contact with each Rommeck et al., 2009b other for a limited period of time and then infants are housed singly during the rest of the time Rotational Infants are continuously housed with Novak & Sackett, 1997; Rommeck et al., different peers 2009b Surrogate mothers Inanimate objects are placed into the cage as an Capitanio & Mason, 2000; Dettmer et al., rearing (SMR) artificial surrogate mother 2008; Schneider & Suomi, 1992; Suomi, 1973 Surrogate-peer Combination of SMR and PR Bastian et al., 2003; Lutz et al., 2007; rearing (SPR ) Meyer et al., 1975 Temporary One time Infants are taken away from their mothers at later Hinde & Mcginnis, 1977; Hinde et al., 1966; stages of life for a period of time, followed by mother- Kaufman & Rosenblum, 1967; Seay et al., infant reunion 1962; Spencer-Booth & Hinde, 1971 Repetitive Repeatedly separating infants from their natal group Clarke et al., 1998; Dettling et al., 2002a, b; for relatively short periods of time, followed by Levine & Mody, 2003; Sánchez et al., repeated reunions 2005; Suomi et al., 1983 Maternal neglect Infant-mother was confronted with various foraging Andrews & Rosenblum, 1991; Coplan et conditions to induce different levels of stress in the al., 1996; Rosenblum & Andrews, 1994; mother Rosenblum & Paully, 1984 form of her availability all the time, her physical touching, Previous study showed that continuous rotational pairing caring and protection, which was an obvious fact to us today induces a behavioral profile quite similar with that of mother without any necessity to prove. rearing in socially complex environment (Rommeck et al., 2011). However, although isolation models are important in highlighting Compared with social isolation, PR is less severe and thus the devastating consequences of maternal deprivation, the more widely used in recent NHP EARE studies. Surrogate extreme manipulations could induce severe cognitive and mothers rearing (SMR) is another early rearing method, in emotional deficits, or even self-injurious behaviors, which are which inanimate object is placed into the cage as an artificial very difficult to remediate in primates. Therefore, less severe surrogate mother (Capitanio & Mason, 2000; Dettmer et al., rearing conditions were developed afterwards at least partially 2008; Eastman & Mason, 1975; Harlow, 1958; Harlow & due to ethical considerations. Zimmermann, 1959; Hennessy & Kaplan, 1982; Kaplan, 1974; Mason & Berkson, 1975; Roy et al., 1978; Schneider & Suomi, Maternal separations 1992; Suomi, 1973). Infants could quickly develop attachment Peer-rearing (PR) (or nursery rearing, NR) (including continuous with surrogate mothers, and some studies indicated that the pair rearing, intermittent and rotational peer rearing) is another infants usually preferred cloth surrogate mothers than wired widely used rearing condition, in which infants were reared ones (Harlow, 1958; Harlow & Zimmermann, 1959). Previous together with peers of the same age (Chamove et al., 1973; study reported that surrogate mothers could affect the Erwin et al., 1973; Sackett, 1967; Worlein & Sackett, 1997) behaviors of infants, and different characters of surrogate (Table 1). In continuous pair rearing condition, infants are mothers such as mobility and orientation had different usually reared by pairs throughout development (Chamove et influences (Dettmer et al., 2008). Surrogate-peer rearing (SPR) al., 1973; Fekete et al., 2000; Hotchkiss & Paule, 2003; Novak method is a combination of SMR and PR, in which the infants & Sackett, 1997). Intermittent peer rearing allow peers to are reared with inanimate surrogate mothers (SMR condition) contact with each other for a limited period of time, and then during the initial several months of life, and then are allowed to infants are housed singly during the rest of the time (Rommeck have peer interactions for a limited period of time (PR condition) et al., 2009b). Within the rotational peer rearing condition, (Bastian et al., 2003; Lutz et al., 2007; Meyer et al., 1975). infants are continuously peer housed with different infant Comparing with permanent removal of the mother, infants are partners (Novak & Sackett, 1997; Rommeck et al., 2009b). not separated from their mothers right away at born in Zoological Research 38(1): 7-35, 2017 9 temporary maternal separations, but after a period of time demand have to work hard to get food (Andrews & Rosenblum, usually several hours, days or weeks, following by mother-infant 1991; Coplan et al., 1996; Rosenblum & Andrews, 1994; reunion (Hinde et al., 1966; Hinde & McGinnis, 1977; Kaufman & Rosenblum & Paully, 1984). The advantage of this model is that Rosenblum, 1967; Seay et al., 1962; Spencer-Booth & Hinde, even though infants are still in adverse situation, the severe 1971). Temporary maternal separation usually contains a one- adverse experience of mother and peer deprivation can be time separation although different time delay could be adopted. avoided. In addition, other rearing strategies are applied in this A modified version of one-time separation is repetitive mother- model, i.e., infants were reared by a female which was not their infant separation, in which infants are separated from and biological mother (Maestripieri, 2005; Novak & Suomi, 1991); reunited with their natal group repeatedly for relatively short infants were housed with non-reproductive female adults periods of time (Clarke et al., 1998; Dettling et al., 2002b; (Champoux et al., 1989b). Levine & Mody, 2003; Sánchez et al., 2005; Suomi et al., 1983). The impact of these procedures appeared to be further EARE EFFECTS intensified if the separations were unpredictable (Levine, 2000; Although partial social isolation tends to induce less severe Sánchez et al., 2005). Unlike social isolation, maternal defects than total social isolation, the expression of behavior separation adopted relatively mild manipulations, the presence defects is similar. Isolated monkeys reared without exposure to of surrogate mothers and the opportunity of direct contact with companions during early life, especially the first 6 months, develop mothers and peers added social complexity to the infants’ living a pervasive pattern of abnormalities referred to as the isolation environment, therefore could avoid severe social and emotional syndrome. Mason (Mason, 1968) summarized the syndrome deficits associated with mothers’ absence. under four headings: (1) abnormal posturing and movements, such as rocking; (2) motivational disturbances, such as Maternal neglect excessive fearfulness or arousal; (3) poor integration of motor Compared with isolation and maternal separation methods patterns, such as inadequate sexual behavior; (4) deficiencies described above, maternal abuse and neglect during early life in social communication, such as failure to withdraw after being are more common in humans, therefore are more widely used threatened by an aggressing animal. In this section, the effects on NHPs to study adult mood and anxiety disorders. In NHP of EARE on social behaviors, learning and memory ability, brain maternal neglect models, in order to induce stress in the mother, structural and functional developments, including influences on infant mothers are confronted with various foraging conditions, neurons and glia cells, neuroendocrine dysregulation, such as variable/unpredictable foraging demand (VFD), especially stress related HPA axis will be reviewed. consistently low foraging demand (LFD) and consistently high (but predictable) foraging demand (HFD). Mothers in LFD Social behavior condition have easy access to food while those in high foraging Effects of EARE on social behaviors are detailed in Table 2. Table 2 Effects of EARE on social behaviors Behavior types Behavior descriptions Stereotypic behaviors Whole-body stereotypes (e.g., rocking, pacing, bouncing, swing, and back-flipping) Self-directed stereotypes (e.g., saluting, digit-sucking, self-clutching, self-clasping, eye-poking, eye-covering and hair-pulling) Self-directed behaviors Self-manipulation, self-scratching, self-grasping, self-rubbing Self-injurious behavior (SIB) Aggression Less aggression during infancy and more aggression during later life Affiliative behavior Tend to show more affiliative behavior during infancy but less affiliative behavior during adulthood Social and environmental exploration Decreased social and environmental exploration Social dominance Tend to show low dominance rank Sexual behaviors Less and abnormal sexual behaviors Others Polyphagia and polydipsia Harlow, 1962; Harlow & Suomi, 1971a; Mason & Sponholz, Disturbance behavior 1963; Mitchell, 1968; Suomi et al., 1971). Among these Monkeys exposed to adverse early experience tended to show monkeys, some abnormal movements, such as rocking and more disturbance behaviors, such as stereotypic and self- self-grasping, could present very early in their lives, even at the directed behaviors, motivational disturbances and social first month (Baysinger et al., 1972). Additionally, some of these deficiency. The isolated monkeys appeared to show more behaviors could turn into stereotypic behaviors, including disturbance behaviors, including crouching, clutching, rocking, repetitive movements or postures, as well as ritualized pacing, flipping, hugging, clasping, thumb-sucking (Harlow & movements, and could be divided into whole-body stereotypes 10 www.zoores.ac.cn (e.g., rocking, pacing, bouncing, swing, and back-flipping), self- compared with MR infants. Lack of sufficient social interaction directed stereotypes (e.g., saluting, digit-sucking, self-clutching, led to the fact that EARE exposed monkeys could not self-clasping, eye-poking, eye-covering and hair-pulling) and successfully adapt to living in a large social group (Griffin & other idiosyncratic behaviors (e.g., teeth grinding, head tossing, Harlow, 1966; Harlow & Harlow, 1962; Mason & Sponholz, or making noise by blowing air into the cheeks). It was reported 1963; Ruppenthal et al., 1991). Not only social interaction, that whole-body stereotypes were much more common than studies also showed decreased environmental exploration in self-directed stereotypes (Lutz et al., 2003). Previous studies isolated monkeys (Griffin & Harlow, 1966; Mason & Sponholz, indicated that isolated monkeys showed more repetitive whole-body 1963; Mitchell, 1968), VFD and PR monkeys (Rosenblum & stereotypes (Mitchell, 1968), while PR monkeys showed more self- Paully, 1984; Ruppenthal et al., 1991). Another major index of directed stereotypes (Lutz et al., 2003; Suomi et al., 1971). exploratory behavior is locomotor activity, while some NHP EARE exposed monkeys tended to show more self-directed studies showed less locomotion in isolated adults (Harlow & behaviors. Isolated monkeys showed self-manipulation, self- Suomi, 1971a; Mason & Sponholz, 1963; Mitchell, 1968) and scratching, self-grasping, self-rubbing, and autoeroticism while PR infants (Feng et al., 2011), others found no differences in in isolation (Baysinger et al., 1972), or showed remarkable PR adults (Winslow et al., 2003), or even higher activity levels increases in self-clasping soon after removal from isolation in PR infants during the first month after isolation (Champoux et (Harlow et al., 1965; Suomi et al., 1974), or self-clutching after al., 1991). Therefore, there was no agreed tendency of EARE surrogate mother removing (Harlow & Zimmermann, 1959). PR influence on locomotor activity in monkeys, making it an invalid reared infants and juvenile monkeys showed increased self- measure of exploratory behavior if used alone (Wright, 1983). stimulation behaviors, including self-sucking, self-clinging, self- Another domain of EARE induced social deficiency is social clasping and other self-directed behaviors (Champoux et al., dominance. In monkey society, social dominance is a complex 1991; Lutz et al., 2003; Suomi et al., 1971). Moreover, short- phenomenon mediated by different mechanisms and various term stress by temporary physical restrictions could also induce factors such as kinship, age, sex, and physical factors like body significant increases in self-clasping and huddling behaviors weight, appearance and health (Bernstein & Cooper, 1999; when the infants returned to their home cages (Harlow & Suomi, Bernstein & Mason, 1963; Morgan et al., 2000; Sprague, 1998; 1971a). Those self-directed behaviors often turned into self- Takahashi, 2002). Kinship seemed to be the major factor in injurious behavior (SIB), with males showing a much higher determining dominant rank at least until puberty (Koford, 1963; level of vulnerability than females (Cross & Harlow, 1965; Koyama, 1967), but became weaker during the development Gluck & Sackett, 1974; Lutz et al., 2003; Suomi et al., 1971), (Bernstein & Williams, 1983). Both dominance formation and and PR monkeys usually much more vulnerable than MR maintenance among males in a living group are usually monkeys (Rommeck et al., 2009a). Surrogate mothers achieved by aggressive behavior such as fighting, with the appeared to provide a certain degree of contact acceptability, stronger and more aggressive subjects winning and thus security and trust sufficient for isolated monkeys to suppress becoming dominant. However, appropriate use of aggression is existing self-directed disturbance activity, and to initiate crude critical for both acquiring and maintaining social status, as social interactions with other isolated monkeys (Harlow & overly aggressive monkeys may risk social ostracism from their Suomi, 1971b). However, Lutz et al. (Lutz et al., 2007) reported conspecifics. Moreover, aggressive behavior was not that SPR monkeys showed significantly more self-biting indispensable to obtain and keep dominance status and comparing to PR and MR reared animals, and it was suggested dominance sustained without aggression was more stable than that surrogate rearing in combination with lower levels of social that formed on the basis of aggression (Fonberg, 1988). contact during play may be risk factors for the later Monkeys exposed to EARE tended to show less aggression development of self-biting behavior. Actually, self-directed during infancy (Chamove et al., 1973; Harlow et al., 1965), and behaviors were hypothesized to result from the redirection of more aggression during later life (Chamove et al., 1973; normal social behaviors toward one's own body and were Mitchell, 1968; Suomi et al., 1974; Winslow et al., 2003). The suggested to be symptoms of some mental diseases (Goosen, aggressive monkeys exposed to EARE may repeatedly attack a 1981; Mason & Berkson, 1975). These findings indicate that helpless infant or attempt to attack a dominant male, while EARE exposed monkeys could be used as an ideal model of infant-directed aggression is abnormal adult-directed aggression is related human mental disorders from behavioral perspective. both abnormal and suicidal (Chamove et al., 1973; Mitchell, 1968; Suomi et al., 1974; Winslow et al., 2003). On the other Social deficiency hand, studies showed EARE exposed monkeys showed In natural environments, infants and juvenile monkeys are heightened fear in all age stages (Champoux et al., 1991; supposed to be more active in joining the social play with peers, Dettling et al., 2002b; Levine & Mody, 2003; Mitchell, 1968). It but monkeys exposed to EARE show decreased social playing. seems that EARE makes monkeys more emotional in two Isolated monkeys showed less (Harlow et al., 1965; Mitchell, opposite directions, both aggression and fear. In addition to 1968), or even no contact playing at all (Harlow et al., 1965). aggression, affiliative behavior, such as grooming and proximity, Pair and peer reared infants (Chamove et al., 1973), VFD is also important in establishing and maintaining alliances and reared infants (Andrews & Rosenblum, 1991; Rosenblum & reinforcing the dominance hierarchy. Affiliative behavior was Paully, 1984), repeated parental deprivation infants (Dettling et suggested to be more positively related to dominance rank than al., 2002b; Levine & Mody, 2003) all showed less social playing kinship in Japanese monkeys (Singh et al., 1992). On the Zoological Research 38(1): 7-35, 2017 11 contrary to aggression, EARE exposed monkeys showed more aggressive, rank-related, sexual, and emotional behavior in affiliative behavior during infancy (Chamove et al., 1973; rhesus monkeys (Bauer & Baker, 2016). Rosenblum & Paully, 1984; Ruppenthal et al., 1991), but less affiliative behavior during adulthood (Kraemer & McKinney, 1979; Learning and memory Rosenblum & Paully, 1984; Winslow et al., 2003). With more Early primate studies showed EARE exposed adults performed aggressive and less affiliative behavior which both contribute to adequately on simple discriminations or delayed-response acquiring and reinforcing social dominance, EARE exposed adult (Gluck et al., 1973), but showed impairments in certain complex monkeys are supposed to have low social dominant rank in a tasks such as those requiring engaging working memory with living group, and studies indeed indicated that both isolated and dynamic rules or delays or response inhibition (Beauchamp & PR adult monkeys showed low social dominance (Kraemer & Gluck, 1988; Beauchamp et al., 1991; Gluck et al., 1973; Gluck & McKinney, 1979; Mitchell, 1968; Ruppenthal et al., 1991). Sackett, 1976; Sánchez et al., 1998). These results were obtained mostly from adult monkeys separated from their Sexual behavior mothers at birth and reared in total isolation for 9-12 months. Monkeys exposed to EARE demonstrated less or abnormal PR reared juvenile monkeys also showed cognitive deficits, sexual behaviors (Chamove et al., 1973; Harlow et al., 1966; they had more difficulty acquiring the delayed non-matching to Harlow, 1962; Harlow et al., 1965; Mitchell, 1968). Abnormal sample (DNMS) task and were also impaired in object but not sexual behaviors (abortive mount) is defined as any improperly spatial reversal learning (Sánchez et al., 1998). Moreover, even oriented mount, accompanied by pelvic thrusting including brief social isolation impaired performance in a multiple video- standing-to-head, standing-to-side and ventral lie-on (Wallen et task assessment in adult rhesus monkeys (Washburn & al., 1981). Males usually were not mount properly as they Rumbaugh, 1991) and impaired reversal learning and engaged in varied but misplaced heterosexual efforts, while behavioral inhibition in adult marmosets (Pryce et al., 2004a, b). females were not maintain the sexual present (stood These results were consistent with the results of human studies, quadripedally with the perineal area directed towards the which showed the post institutionalized children (Bauer et al., recipient) or turned their bodies when mounted. Mount behavior 2009) and childhood exposed to neglect and abuse (Majer et al., includes no-foot-clasp mount and foot-clasp mount, which could 2010) were associated with impaired learning and memory be differentially affect by different EARE. Males with short during adulthood. Although those studies revealed EARE access periods with peers (0.5 h) rarely or never foot-clasp- induced impairment of learning and memory ability in a task mounted peers, while those given 24 h access regularly foot- dependent way in adult monkeys, other primate studies indicate clasp-mounted peers (Wallen et al., 1981). Isosexually exposure to mild early life stress improves prefrontal dependent reared males showed less foot-clasp mounting and more response inhibition in primates, suggesting its beneficial effect presenting than heterosexual males, while conversely, on cognitive control (Parker et al., 2005, 2012). isosexually reared females showed statistically more mounting and less presenting than heterosexual females Brain structure and function (Goldfoot et al., 1984). Moreover, females exposed to EARE The first documentation of the effects of negative early also showed abnormal maternal behaviors, in a way that experiences on monkey brain was provided by Martin et al. those never experienced mother caring not only were unable (1991), which showed significant alterations in the chemo to exhibit caring to their own offspring, but also far more architecture of the striatum 19-24 years after social deprivation. likely to display inadequate, abusive or neglectful behavior Additionally, Siegel et al. (1993) demonstrated that early social toward their offspring (Bridges et al., 2008; Champoux et al., deprivation resulted in an increase in the amount of non- 1992; Harlow & Suomi, 1971b; Seay et al., 1964; Suomi, phosphorylated neurofilament protein in hippocampal dentate 1978; Suomi et al., 1974; Suomi & Ripp, 1983), consistent gyrus granule cells in rhesus monkeys. Further studies showed with human findings showing abusive behavior appeared to structure and function changes in many brain regions including be transmitted across generations (Roustit et al., 2009). amygdala, hippocampus, prefrontal cortex (PFC), anterior Primate studies also showed other EARE induced behavioral cingulate cortex (ACC), corpus callosum and cerebellum etc, effects besides listed above, including polyphagia and both in humans and animals exposed to EARE (Andersen, polydipsia in isolated adults (Miller et al., 1969), more 2015; Bick & Nelson, 2016; Gilmer & McKinney, 2003; Gorman vulnerable to excessive alcohol consumption (Fahlke et al., et al., 2002; Hart & Rubia, 2012; Korosi et al., 2012; McEwen, 2000; Higley et al., 1991) and elevated response to both 2003; Worlein, 2014)(Table 3). aversive and rewarding stimuli (Nelson et al., 2009) in PR monkeys and abnormal sleep rhythmicity (Barrett et al., 2009; Amygdala Boccia et al., 1989; Kaemingk & Reite, 1987; Reite et al., 1974; Amygdala is a group of almond-shaped nuclei located deep Reite & Short, 1978). An interesting research showed EARE within the medial temporal lobes of the brain in complex significantly influenced the development of lateralisation, as PR vertebrates. It was considered as the emotion center and monkeys demonstrated greater left-hand bias compared to MR responsible for emotion reactions like reward, fear and anxiety reared monkeys (Bennett et al., 2008). Despite of EARE effects (Davis, 1992; Gallagher & Chiba, 1996; Ledoux, 2003; Phelps, described above, recent research suggested that modern PR 2006). Rodent studies showed acceleration of amygdala practices might not result in inevitable perturbations in development in early weaning rodents (Kikusui & Mori, 2009; 12 www.zoores.ac.cn Table 3 Effects of EARE on brain structure and function Outcomes Human Studies Primate Studies Amygdala Children No significant volumes changes De Bellis et al., 2001; De Brito et al., 2013; No significant volume changes (Howell Hanson et al., 2010; Woon & Hedges, 2008 et al., 2014); Larger volume and elevated Lupien et al., 2011; Mehta et al., 2009; Decreased SERT binding potential response Tottenham et al., 2010 (Ichise et al., 2006); Decreased volume Edmiston et al., 2011; Hanson et al., 2015; Differential expression of one gene Luby et al., 2013 GUCY1A3 (Sabatini et al., 2007) Adults No significant volume changes Bremner et al., 1997; Cohen et al., 2006 Larger volume Evans et al., 2016; Lyons-Ruth et al., 2016 Elevated activity Casement et al., 2014; Javanbakht et al., 2015; Kim et al., 2013 Hippocampus Children Decreased volume Edmiston et al., 2011; Hanson et al., 2015; No significant volume change (Law et Luby et al., 2013 al., 2009a, b; Sánchez et al., 1998; No significant volume change Carrion et al., 2001; De Bellis et al., 2001; Spinelli et al., 2009) De Bellis et al., 1999; De Bellis et al., 2002; Mehta et al., 2009; Tottenham et al., 2010; Woon & Hedges, 2008 Adults Decreased volume Bremner et al., 1997; Cohen et al., 2006; Stein et al., 1997; Woon & Hedges, 2008 Prefrontal Children No significant volume changes De Bellis et al., 1999 Greater enlarged medial prefrontal cortex (PFC) cortex (mPFC) size (Spinelli et al., De Bellis et al., 2002; Edmiston et al., 2011; 2009) Decreased volume Hanson et al., 2010; Morey et al., 2016; Thomaes et al., 2010 Larger volume Carrion et al., 2009; Richert et al., 2006 Tomoda et al., 2009; van Harmelen et al., Decreased volume 2010 Casement et al., 2015; Kim et al., 2013; Reduced activity Adults Romens et al., 2015; Schweizer et al., 2016 Casement et al., 2014; Javanbakht et al., Increased response 2015; Jedd et al., 2015; Wang et al., 2016; White et al., 2015 Ono et al., 2008). The limited amount of primate studies found found no significant changes of amygdala volume (Bremner et no significant amygdala volume changes (Howell et al., 2014), al., 1997; Cohen et al., 2006), while others found larger volume but functional changes including decreased SERT binding (Evans et al., 2016; Lyons-Ruth et al., 2016), interrupted potential (Ichise et al., 2006) and differential expression of one regulation of negative emotion (Kim et al., 2013), increased gene GUCY1A3 (Sabatini et al., 2007) in amygdala of EARE response to potential rewards (Casement et al., 2014), elevated exposed monkeys. However, human studies in maltreated amygdala responses to threat but not happy faces (Javanbakht children showed contrary results, with some studies found no et al., 2015). In addition to amygdala structure and activity volume changes (De Bellis et al., 2001; De Brito et al., 2013; changes, its connectivity with other brain regions was also Hanson et al., 2010; Woon & Hedges, 2008), while others affected (Barch et al., 2016; Jedd et al., 2015). Despite those revealed decreased volume (Edmiston et al., 2011; Hanson et controversial results, the influence of EARE on emotion such as al., 2015; Luby et al., 2013) or greater volume and elevated the elevated response to emotion stimuli both in human and response (Lupien et al., 2011; Mehta et al., 2009; Tottenham et primates (Casement et al., 2014; Javanbakht et al., 2015; al., 2010). Furthermore, those studies found greater volume Nelson et al., 2009) should be mainly achieved through its and elevated response of amygdala (Mehta et al., 2009; influence on amygdala. Tottenham et al., 2010) were performed several years after the institutionalized children adopted by high socio-economic status Hippocampus families. These data suggested that EARE modified amygdala Hippocampus, a major component of the brains located inside changes was resistant to recovery, and it was consistent with the medial temporal lobe and beneath the cortical surface, is primate research that suggested abnormal behaviors was involved in episodic, declarative, contextual, and spatial resistant to environmental enrichment treatments (Lutz et al., learning and memory, as well as being a component in the 2004; Lutz & Novak, 2005; Novak et al., 1998; Rommeck et al., control of autonomic and vegetative functions (Buckley, 2005; 2009a). Similarly, in adults exposed to EARE some studies Eichenbaum, 2001; Eichenbaum et al., 1992, 1996; Manns & Zoological Research 38(1): 7-35, 2017 13 Eichenbaum, 2006; Opitz, 2014; Shohamy & Turk-Browne, childhood maltreatment was a consistent finding (Tomoda et 2013). In human studies, EARE induced significant reduction of al., 2009; van Harmelen et al., 2010). That might due to the hippocampal volume was an consistent finding in adults fact that PFC continues to develop during adolescence (Bremner et al., 1997; Cohen et al., 2006; Hart & Rubia, 2012; (Houston et al., 2014; Lenroot & Giedd, 2006; Richards & Xie, McCrory et al., 2011; Stein et al., 1997; Woon & Hedges, 2008). 2015), therefore might be particularly vulnerable to the effects However, children and adolescents studies showed inconsistent of stress during adolescence. In addition to the structural results, with few found decreased volume (Edmiston et al., 2011; changes, EARE could also induce PFC functional changes, Hanson et al., 2015; Luby et al., 2013), while most found no with some human adults exposed to EARE showing reduced significant change (Carrion et al., 2001; De Bellis et al., 2001, prefrontal cortex activity during monetary reward anticipation 1999, 2002; Mehta et al., 2009; Tottenham et al., 2010; Woon & and emotion regulation (Casement et al., 2015; Kim et al., Hedges, 2008). Primate studies also found no significant 2013; Romens et al., 2015; Schweizer et al., 2016), while hippocampal volume change in PR (Sánchez et al., 1998; others showing increased response to potential rewards and Spinelli et al., 2009) and repeated mother deprived (Law et al., threatening faces and in passive viewing conditions 2009b) juvenile monkeys, suggesting changes of hippocampus (Casement et al., 2014; Javanbakht et al., 2015; Jedd et al., seemed to happen later in life compared to early life amygdala 2015; Wang et al., 2016; White et al., 2015). One primate changes. Two possible explanations could account for the report indicated PR juvenile monkeys showed greater discrepancy of children and adult findings. Firstly, that might enlarged medial prefrontal cortex (mPFC) size (Spinelli et al., due to the fact that the hippocampus develops mainly in the first 2009). Moreover, both rodent and primate studies revealed years of life, therefore less affected by exposure to adversity in the direct underlying epigenetic mechanisms of EARE on PFC childhood and adolescence (Houston et al., 2014; Lenroot & through influencing differential gene expression, histone Giedd, 2006; Richards & Xie, 2015). Another possibility is that acetylation and DNA methylation (Blaze et al., 2015a; EARE might not have an immediate effect on the hippocampus Provençal et al., 2012; Wall et al., 2012). Studies regarding but induced changes over time, and long-term effects of EARE EARE effects on PFC in primates are rare, and further exposure may be delayed and became manifest only in later investigations are necessary. phases of development when the vulnerable brain reaches maturation (Andersen & Teicher, 2004; Brunson et al., 2005; Other brain regions Gluckman & Hanson, 2004; Gluckman et al., 2007; Sapolsky et The anterior cingulate cortex (ACC) is the frontal part of the al., 1985). Moreover, human studies found interesting results cingulate cortex, and appears to play a role in a wide variety of concerned with influence of EARE exposure on structure and rational cognitive functions, such as reward anticipation, activity of hippocampus and amygdala, with decreased decision-making, empathy and emotion (Devinsky et al., 1995; hippocampal volume and activity in humans exposed to Drevets et al., 2008). It can be divided anatomically into dorsal adulthood stress (Bremner et al., 2007; Lupien et al., 2007; and ventral components, with dorsal part connected with PFC Rauch et al., 2000) and adults experiencing EARE (Bremner et making its involvement in cognition possible, and the ventral al., 1997; Cohen et al., 2006; Stein et al., 1997; Woon & part connected with amygdala making its involvement in Hedges, 2008), while increased amygdala volume and activity emotion possible (Bush et al., 2000; Morecraft et al., 2007). in humans exposed to adulthood stress (Bremner et al., 2007; Human studies showed reduced volume of adult ACC in people Lupien et al., 2007; Rauch et al., 2000) and adults experiencing with mood disorders (Botteron et al., 2002; Drevets et al., 1997; EARE (Mehta et al., 2009; Tottenham et al., 2010). Although the Yamasue et al., 2003), adults exposed to early life stress (ELS) biological mechanism and meaning of this phenomenon (Cohen et al., 2006) and abuse-related Posttraumatic stress remains unclear, that might contribute to or even be the direct disorder ( PTSD ) (Kitayama et al., 2006; Thomaes et al., 2010) reason for the impaired learning and memory ability (decreased and major depressive disorder (Treadway et al., 2009). On the hippocampal volume and activity related) and elevated contrary, a primate study found enlarged ACC in PR juvenile response to emotional stimuli (increased amygdala volume and monkeys (Spinelli et al., 2009). Moreover, an epigenetic study activity related) described above. showed parental separations in infant marmoset affected expression of genes in the ACC of adolescent monkeys (Law et Prefrontal cortex al., 2009a). Additionally, both human and primate studies The prefrontal cortex (PFC) is the anterior part of the frontal revealed EARE affected cerebellum, with human studies lobes of the brain and implicated in planning complex cognitive showing decreased cerebellum (Bauer et al., 2009; Edmiston et behaviors, personality expression, decision making and al., 2011), while a primate study revealing larger cerebellar moderating correct social behavior. Children and adolescents vermis area in PR juvenile monkeys (Spinelli et al., 2009). studies showed inconsistent results of EARE induced PFC EARE effect on primate cerebellum might due to the fact that structural changes, with findings of either no significant macaque cerebellum has high density of glucocorticoid differences (De Bellis et al., 1999), or significantly smaller receptors (GRs) (Sánchez et al., 2000), which put it particularly volume (De Bellis et al., 2002; Edmiston et al., 2011; Hanson et vulnerable to stress hormones related over stimulation. Striatum al., 2010; Morey et al., 2016; Thomaes et al., 2010) or was another brain region affected by EARE, with increased significantly larger volume (Carrion et al., 2009; Richert et al., response to potential rewards (Casement et al., 2014) and 2006). In contrast, decreased PFC volume in adults exposed to elevated dopamine responses to amphetamine (Oswald et al., 14 www.zoores.ac.cn 2014), and a potential neurobiological mechanism linking early- numbers in many brain regions, including PFC, mPFC, life adversity and altered ventral striatal development was hippocampus, cingulate cortex and amygdala (Leventopoulos indicated (Goff & Tottenham, 2015). In addition to those specific et al., 2007), and could alter behavioral, autonomic and regional changes, PR chimpanzees showed less global white- endocrine responses to environmental challenge (Musholt et al., to-grey matter volume and cortical folding (Bogart et al., 2014). 2009; Rüedi-Bettschen et al., 2006). Although there was no direct Structural connectivity between different brain regions was also evidence pointing out that glia cell changes were responsible affected by EARE, as studies showed affected corpus callosum, for those altered responses in rats, those studies at least a wide and flat bundle of axons beneath the cortex connecting suggested the involvement of glia cell in EARE induced effects. left and right cerebral hemispheres and facilitating inter- Moreover, human studies showed that glial cell depletion in hemispheric communication, in a inconsistent way that most many brain regions was related to mood disorders, as the human studies showing EARE reduced corpus callosum (De number of glia cell was reduced in PFC of both major Bellis et al., 1999; Rinne-Albers et al., 2016; Teicher et al., 2004, depressive disorder (MDD) and bipolar disorder (BD) patients 1997), while few showing no significant changes (Mehta et al., (Öngür et al., 1998), in the amygdala of major depressive 2009). Primate studies also found either decreased corpus disorder patients (Bowley et al., 2002) and in anterior cingulate callosum size (Sánchez et al., 1998) or no significant changes cortex of major depressive disorder and schizophrenia patients (Spinelli et al., 2009). (Cotter et al., 2001). Considering the important trophic influence of glia on neurons, glia cell deficits induced by EARE could Neurons and glia cells possibly be responsible for EARE effects on neurons and Neurons are the basic unit of brain. Neuronal network is furthermore to abnormal behavioral function. If that is true, how responsible for the daily cognitive and emotional behaviors. Glia does it happen? Rodent Studies showed that stress related cell is a group of non-neuronal cells that support and protect the hormone glucocorticoid receptors (GRs) were also expressed in neurons in the brain. Rodent studies showed that maternal glia cells (Bohn et al., 1991; Jung-Testas & Baulieu, 1998; separation could induce morphological alteration of the apical Vielkind et al., 1990). Glucocorticoid is the product of the HPA dendrites of CA3 pyramidal neurons (Kwak et al., 2008); could axis, so EARE might take effects through its influence on stress increase corticotropin releasing factor (CRF)-containing neurons related hormones, like glucocorticoid, and then exert influence in amygdala (Becker et al., 2007); and could decrease in vivo on glia cells leading to various effects (Jauregui-Huerta et al., firing activity of amygdala neurons (Adams & Rosenkranz, 2016) 2010). Indeed, in vitro and in vivo studies showed that and sex related neurogenesis (Oomen et al., 2009). Chronic glucocorticoids could influence gene expression in glia cells stress could induce atrophy of dendrites in hippocampus of rats (Bohn et al., 1994; Kumar et al., 1985) and could regulate the (Brunson et al., 2005; McEwen, 1999) and tree shrews concentration of glial fibrillary acidic proteins (O'Callaghan et al., (Magariños et al., 1996), and could induce hippocampal 1989). By playing central roles in learning and memory, neuroplasticity changes (Fenoglio et al., 2006). Bartesaghi and hippocampal astrocyte number was dose-dependently colleagues used guinea-pig as animal model to investigate the increased by corticosterone treatment (Bridges et al., 2008), effects of early isolation on neurons, and they found that early and glial responses in hippocampus was also regulated by isolation could induce morphologic changes of neurons in glucocorticoid through influencing gene expression (Nichols et entorhinal cortex and hippocampus (Bartesaghi et al., 2003a, b; al., 2005). However, few studies were performed to investigate Bartesaghi & Serrai, 2001, 2004). Although primate studies this issue and EARE affected glia cell changes were link directly found neuronal morphological changes in EARE exposed to behavioral outcomes without solid evidence. In primate monkeys (Bryan & Riesen, 1989; Floeter & Greenough, 1979; studies, there are lack of evidence to support that EARE affects Stell & Riesen, 1987; Struble & Riesen, 1978), these early glia cell structural and functional changes, and furthermore, findings were limited to cerebellum, somatosensory and motor induces behavioral outcomes. cortex, with limited information on other important brain regions, such as hippocampus, amygdala and PFC. Recent studies Lateralisation showed that different environments could induce neuron Some studies suggested that the influence of EARE on different plasticity changes in the key brain regions involved in learning brain hemisphere might be different, and different type of EARE and memory. Complex environment could enhance complexity might take effects differentially on the same brain structure. A of the dendritic tree and density of dendritic spine in human study found that the institutionalized children had hippocampus and PFC in monkeys (Kozorovitskiy et al., 2005). greater right amygdala volume, while the left amygdala volume Early parental deprivation in the marmoset monkey could was smaller in the children experienced longer periods of produce long-term changes in hippocampal expression of deprivation (Mehta et al., 2009). Another human study showed genes involved in synaptic plasticity and implicated in mood that patients with child abuse-related complex PTSD showed disorder (Law et al., 2009b). So these neuron morphological reduced gray matter concentration in right hippocampus and and plasticity changes might explain and account for how right dorsal ACC, but not in the left areas (Thomaes et al., EARE take effects on cell level, and then further more leading 2010). In primate studies, maternal separation was associated to behavioral changes. with activation in the right dorsolateral PFC and decreased As EARE effects on glia cells, rodent studies revealed that activity in the left dorsolateral PFC of juvenile rhesus monkeys EARE could induce long-term changes of astrocyte density and (Rilling et al., 2001). Not only brain structure and function Zoological Research 38(1): 7-35, 2017 15 showed lateralisation affection by early experiences, behavioral decreasing in number and length of apical dendritic branch of research also found lateralisation in primates, as an interesting medial prefrontal cortex in male rats, whereas increasing in research showed that PR monkeys demonstrated greater left- apical dendritic length in female rats (Garrett & Wellman, 2009). hand bias compared to MR reared monkeys (Bennett et al., Isolated males showed less dendritic branches, shorter 2008). The number of lateralisation related studies is limited dendritic length and smaller dendritic spine density than control and the underlying mechanism remains unknown, which males, while isolated females had more dendritic branches than certainly adds complexity to the understanding of the influence control females in guinea pig (Bartesaghi et al., 2003a). EARE on brain structure and functional changes and the Neurogenesis was significantly increased in male but related abnormal behavioral outcomes. decreased in female offspring after maternal deprivation in rats (Oomen et al., 2009). The mechanism of those sexual Other EARE effects differences remains unclear, but one possible explanation is the Young animals are particularly vulnerable to EARE effects gender related physiological differences, such as neuroendocrine Adverse experience has its influence over all life stages, system and brain structure and function, which may induce including early, middle and later life, in which infants are different behavioral and physiological responses in male and especially vulnerable to EARE and the consequences could female subjects. . persistent into later life. That might due to the fact that the most sensitive period of the whole life is the early stage, during which Time effects of EARE the body is undergoing profound physiological development, Early life is a time of heightened susceptibility to EARE and such as HPA axis, and brain is also undergoing profound neural expression of adverse experiences induced effects would be development, such as neurogenesis. The amygdala developes different across life time, therefore the time of administration of rapidly during the early postnatal period in animals, e.g., in rats, adverse experiences and subjects age of measurement might cats and primates (Kikusui & Mori, 2009; Lupien et al., 2009; partially explain the discrepant findings across studies Payne et al., 2010; Wakefield & Levine, 1985). Stress related (Tottenham & Sheridan, 2009). hormones and receptors were also maximally expressed in the The time of adverse experiences administration is important, brain early in development (Avishai-Eliner et al., 1996; Baram & as different brain regions might have unique windows of Hatalski, 1998; Meaney & Szyf, 2005; Pryce et al., 2005a; vulnerability to stress, e.g., human studies indicate that the time Vazquez et al., 2006). These early physiological development window of hippocampus, corpus callosum and frontal cortex is heighten the vulnerability of the brain to environmental at ages of 3-5, 9-10 and 14-16 years, respectively (Andersen et exposures. On the other hand, the proper development needs al., 2008). Rodent studies revealed the critical importance of proper environmental stimuli, and the natural and best stimuli specific time windows early in life for the outcome of maternal during early life is the attachment between caregivers, separation (Bock et al., 2005; Gos et al., 2008; Pryce et al., especially mothers, and infants, as mothers could supply tactile 2005b). Early primate studies by Harlow et al. showed the contact, physical warmth, nourishment, and psychological importance of administration time of adverse experiences comforts. As stated in attachment theory and affectional system, (isolation), in a way that isolation beginning at birth generated infants need to develop a stable relationship with the mother for most severe effects and persisting abnormalities (Harlow et al., social and emotional development to occur normally, while 1965; Mitchell, 1968), while the isolation starting until later in life various EARE intervene the forming of the bonds of this relation, would produce less severe effects and persistent abnormalities therefore both short-term and long-term devastating influence (Harlow et al., 1965; Mitchell, 1968). Moreover, different lasting are inevitable. period of EARE also produced different effects even was all initiated at birth, i.e., 3 months isolation only induced reversible Sexual differences in EARE effects debilitating behavioral deficits, while at least six months Humans studies showed that affectability of various mental isolation generated most severe effects and persisting disorders were sex-related during development, with boys abnormalities; 3 months isolation induced least, 6 months showing higher tendencies to develop aggression and novelty isolation induced moderate and 12 months isolation induced seeking behaviors (Farrington & Loeber, 2000) while girls more most severe defects (Griffin & Harlow, 1966; Harlow et al., susceptible to anxiety and depression (Kessler, 2003). 1965; Mitchell, 1968). These studies suggested that both the Additionally, EARE influence might also be sex related, e.g., time point of administration of EARE and the lasting period corpus callosum volume reduction was only found in EARE have different influences on behavioral and biological exposed males (De Bellis et al., 1999). Similarly, animal studies outcomes. also revealed the vulnerability of males to EARE in rodents Human studies showed different, or even contrary effects of (Galea et al., 1997; Kikusui & Mori, 2009) and primates (Clarke, EARE in children and adults, suggesting EARE might induce 1993; Cross & Harlow, 1965; Mitchell, 1968; Rommeck et al., differential outcomes across lifespan. For example, childhood 2009a; Suomi et al., 1971). On the contrary, other studies abuse induced significant reduction of hippocampal volume in showed preference of EARE on females in humans (Heim & adults (Bremner et al., 1997; Cohen et al., 2006; Stein et al., Nemeroff, 2001; Klimes-Dougan et al., 2001), rodents (Hoyer et 1997; Woon & Hedges, 2008) but not in children (Carrion et al., al., 2013; Ziabreva et al., 2003b) and primates (Sánchez et al., 2001; De Bellis et al., 2001, 1999; Woon & Hedges, 2008); 2005). Previous studies showed that stress could induce EARE induced hypercortisolism in children (Essex et al., 2002; 16 www.zoores.ac.cn