The onset of puberty in colony-housed male and female titi monkeys (Plecturocebus cupreus): Possible effects of oxytocin treatment during peri-adolescent development.

Oxytocin has been used to treat neurodevelopmental conditions in adolescent patients but possible effects on reproductive development have not been well investigated. The effects of daily intra-nasal oxytocin treatment (12-18 months of age) on puberty and fertility were studied in colony-housed, male and female titi monkeys (Plecturocebus cupreus). Body weight, urinary conjugated pregnanes and estrogens (defining cyclicity) in females, and androgens and sperm in urine of in males, were measured from 1 to 3 years of age to detect puberty. Serum testosterone was also measured in males at 13, 23 and 33 months of age and hemi-castration at 3 years of age enabled assessment of testicular morphometry and oxytocin receptor expression. An oxytocin treatment*time interaction suggested a minor, transient suppression in weight gain after treatment ended. Note that females weighed 10% less across all ages. Oxytocin-treated females exhibited early, spurious ovulations but neither regular cyclicity (≈30 months) nor pregnancies were affected by treatment. Oxytocin did not affect the pubertal increase in urinary androgen or the first appearance of sperm, which occurred as early as 15 months of age. Treatment did delay the puberty-associated rise in serum testosterone in males. All males were pubertal by 22 months and all females by 32 months of age. Although no major male or female fertility outcome was observed, oxytocin demonstrated some physiological effects through a delay of testosterone secretion in males, induction of precocious ovulation in females, and a suppression of general weight gain for the months following treatment.

Social stability influences the association between adrenal responsiveness and hair cortisol concentrations in rhesus macaques.

Hair cortisol concentrations are increasingly being used in both humans and nonhuman animals as a biomarker of chronic stress. However, many details regarding how hair cortisol concentrations relate to the dynamic activity and regulation of the HPA axis are still unknown. The current study explores 1) how the regulation of the HPA axis in infancy relates to hair cortisol concentrations (HCC) in infancy 2) whether this relationship persists into adulthood under conditions of social stability, and 3) how social instability impacts these relationships. All subjects were rhesus monkeys housed in large social groups at the California National Primate Research Center, and all had participated in a 25-hr. long BioBehavioral Assessment (BBA) at 3-4 months of age when four plasma samples were taken to assess HPA regulation, in particular cortisol responses to 1) 2-hour social separation and relocation, 2) sustained challenge, 3) dexamethasone and 4) ACTH administration. In Study 1, hair samples were collected at the end of the BBA testing from 25 infant rhesus monkeys from 2 different stable social groups. In Study 2, hair samples were obtained at three timepoints from 108 adults from 3 different stable social groups (1 in the Spring/Summer and 2 in the Fall/Winter) to examine the temporal stability of the relationship between HCC and HPA axis regulation. In Study 3, subjects included 31 infants and 33 adults from a single social group experiencing social instability following the same procedures as in Studies 1 and 2. Generalized linear models were used to determine if infants' HPA axis activity and regulation predicted HCC in infancy (Study 1), in adulthood with animals living in stable social conditions (Study 2) or in animals living in an unstable social group (Study 3). Results indicated that for both infants and adults living in stable social groups, HCC are associated with the adrenal response to ACTH in infancy. Samples collected in the winter also had higher HCC than those collected in summer. In the unstable social group, adult hair cortisol levels were higher than in the stable social groups. Additionally, there were no consistent relationships between HCC and infant HPA axis regulation among adults or infants living in a group experiencing social instability. These results suggest that the aspects of the HPA axis that drive HCC may differ depending on context. Under stable, non-stressed conditions there seems to be a trait-like association between adrenal responsivity and HCC in infancy and adulthood. However, this association may be reduced or eliminated under conditions of social stress.

Examining cortisol rhythmicity and responsivity to stress in children with Tourette syndrome.

BACKGROUND: Tourette syndrome (TS) is characterized by motor and vocal tics, which are often exacerbated by stress. The hypothalamic-pituitary-adrenocortical (HPA) axis, a major stress response system is thus of interest for understanding TS. METHODS: Diurnal cortisol rhythms were estimated in medication-free children 7-13 years with TS (N=20) and healthy age-matched controls (N=16). Salivary samples were collected on 3 consecutive days from the home. HPA responsivity was assessed by examining cortisol in response to a mock and real MRI scan. RESULTS: The results of diurnal rhythmicity revealed a trend showing marginally lower evening cortisol for the TS group. By contrast, the TS group had higher cortisol levels in response to the stressor. There were strong, negative correlations between evening cortisol and tic severity as well as diurnal cortisol and anxiety. CONCLUSIONS: The children with TS showed increased cortisol in response to the MRI environment, supporting a model of enhanced HPA responsivity. The lower evening cortisol may be the result of chronic daily stress. Alternatively, the negative associations between cortisol and reported anxiety and tics may reflect biologically based anxiolytic properties of tic expression. Taken together, the results clearly implicate involvement of the HPA axis in the neuropathology of TS.

Dimensions of response to novelty are associated with social engagement and aggression in adult male rhesus macaques (Macaca mulatta).

In this study, the authors tested the hypothesis that behavioral response across social and nonsocial, novel and familiar conditions may be guided by the same trait(s) related to impulsivity in adult male rhesus macaques. The authors assessed 23 individuals' behavioral response to a series of nonsocial novel scenarios, as well as aggression and sociality within familiar and novel social contexts. Factor analysis of responses to nonsocial novelty identified two factors: Caution, which reflected latency to engage different novel situations, and Interest in Novelty, which consisted of duration and quality of exploration. Each dimension was associated with different social manifestations. Caution was negatively correlated with social aggression in novel and familiar social circumstances; Interest in Novelty was positively associated with social engagement in familiar, but not novel, social circumstances. The authors conclude that traits influencing impulsive response to novelty contribute to risky and normal social behavior across social contexts.

The effects of morphine, naloxone, and κ opioid manipulation on endocrine functioning and social behavior in monogamous titi monkeys (Callicebus cupreus).

The µ opioid receptor (MOR) and κ opioid receptor (KOR) have been implicated in pair-bond formation and maintenance in socially monogamous species. Utilizing monogamous titi monkeys (Callicebus cupreus), the present study examined the potential role opioids play in modulating the response to separation, a potent challenge to the pair-bond. In Experiment 1, paired male titi monkeys were separated from their pair-mate for 30-min and then received saline, naloxone (1.0mg/kg), morphine (0.25mg/kg), or the KOR agonist, U50,488 (0.01, 0.03, or 0.1mg/kg) in a counter-balanced fashion, immediately prior to a 30-min reunion with their mate. Blood samples were collected immediately prior to and after the reunion. Males receiving morphine approached females less, initiated contact less, and females broke contact with the males less. The increase in cortisol in response to naloxone was greater compared to vehicle, and the increase in cortisol in response to the high dose of U50,488 compared to vehicle approached significance. In Experiment 2, paired males were treated with the KOR antagonist, GNTI (0.1, 0.3, or 1.0mg/kg), or saline 24h prior to a 60-min separation from their mate. Blood samples were collected at the time of injection and immediately before and after separation. Administration of the low dose of GNTI decreased the locomotor component of the separation response compared to vehicle. The present study found that the opioid system is involved in both the affiliative and separation distress components of a pair-bond, and these components are regulated by different opioid receptors.

µ and κ opioid receptor distribution in the monogamous titi monkey (Callicebus cupreus): implications for social behavior and endocrine functioning.

The opioid system is involved in infant-mother bonds and adult-adult bonds in many species. We have previously shown that µ opioid receptors (MORs) and κ opioid receptors (KORs) are involved in regulating the adult attachment of the monogamous titi monkey. The present study sought to determine the distribution of MOR and KOR in the titi monkey brain using receptor autoradiography. We used [(3)H][D-Ala(2),N-Me-Phe(4),Gly(5)-ol]-enkephalin (DAMGO) to label MORs and [(3)H]U69,593 to label KORs. MOR binding was heterogeneous throughout the titi monkey brain. Specifically, MOR binding was observed in the cingulate gyrus (CG), striatum, septal regions, diagonal band, amygdala, hypothalamus, hippocampus, and thalamus. Binding was particularly dense in the septum, medial amygdala, paraventricular nucleus of the hypothalamus, mediodorsal thalamus with moderate binding in the nucleus accumbens. Consistent with other primate species, MOR were also observed in "neurochemically unique domains of the accumbens and putamen" (NUDAPs). In general KOR binding was more homogenous. KORs were primarily found in the CG, striatum, amygdala and hippocampus. Dense KOR binding was observed in the claustrum. Relative MOR and KOR binding in the titi monkey striatum was similar to other humans and primates, but was much lower compared to rodents. Relative MOR binding in the titi monkey hypothalamus was much greater than that found in rodents. This study was the first to examine MOR and KOR binding in a monogamous primate. The location of these receptors gives insight into where ligands may be acting to regulate social behavior and endocrine function.

Increased social fear and decreased fear of objects in monkeys with neonatal amygdala lesions.

The amygdala has been implicated in the mediation of emotional and species-specific social behavior (Kling et al., 1970; Kling and Brothers, 1992; Kluver and Bucy, 1939; Rosvold et al., 1954). Humans with bilateral amygdala damage are impaired in judging negative emotion in facial expressions and making accurate judgements of trustworthiness (Adolphs et al., 1998, 1994). Amygdala dysfunction has also been implicated in human disorders ranging from social anxiety (Birbaumer et al., 1998) to depression (Drevets, 2000) to autism (Bachevalier, 1994; Baron-Cohen et al., 2000; Bauman and Kemper, 1993). We produced selective amygdala lesions in 2-week-old macaque monkeys who were returned to their mothers for rearing. At 6-8 months of age, the lesioned animals demonstrated less fear of novel objects such as rubber snakes than age-matched controls. However, they displayed substantially more fear behavior than controls during dyadic social interactions. These results suggest that neonatal amygdala lesions dissociate a system that mediates social fear from one that mediates fear of inanimate objects. Furthermore, much of the age-appropriate repertoire of social behavior was present in amygdala-lesioned infants indicating that these lesions do not produce autistic-like behavior in monkeys. Finally, amygdala lesions early in development have different effects on social behavior than lesions produced in adulthood.

Generic aspects of primate attachments: parents, offspring and mates.

We examine behavioral and physiological aspects of primate emotional attachments in the context of four relationships: infant-to-parent, parent-to-infant, and adult male-to-female and adult female-to-male in a monogamous New World species. Emotional attachments in each of these relationships show striking similarities at a basic functional level. The nature of these similarities suggests that they are produced by the same psychoneuroendocrine core, which appears to be present in all mammals. We also consider the development of each of kind of attachment. In contrast to fundamental similarities in the expression of attachment, their development in each case appears to be based on distinct, species-typical dispositions and constraints.

The effects of bilateral lesions of the amygdala on dyadic social interactions in rhesus monkeys (Macaca mulatta).

The role of the amygdala in dyadic social interactions of adult rhesus monkeys (Macaca mulatta) was assessed after bilateral ibotenic acid lesions. Social, nonsocial, and spatial behaviors of amygdalectomized and control monkeys were assessed in 3 dyadic experiments: constrained, unconstrained, and round robin. Lesions produced extensive bilateral damage to the amygdala. Across all experiments, the amygdalectomized monkeys demonstrated increased social affiliation, decreased anxiety, and increased confidence compared with control monkeys, particularly during early encounters. Normal subjects also demonstrated increased social affiliation toward the amygdalectomized subjects. These results indicate that amygdala lesions in adult monkeys lead to a decrease in the species-normal reluctance to immediately engage a novel conspecific in social behavior. The altered behavior of the amygdalectomized monkeys may have induced the increased social interactions from their normal companions. This is contrary to the idea that amygdalectomy produces a decrease in social interaction and increased aggression from conspecifics.

Breeding readiness in squirrel monkeys: female-primed females are triggered by males.

The South American squirrel monkey (Saimiri) shows pronounced annual variations in reproductive activity. We investigated the contribution of social stimulation to breeding readiness, as reflected in behavior and gonadal hormones. A previous study indicated that formation of new heterosexual pairs can induce breeding readiness in males, but not in females. In the present study we examined the response of individually housed females to the formation of triads of unacquainted females and, subsequently, the response of these triads to the introduction of a single male. The formation of the all-female group was accompanied by a marked reduction in basal cortisol levels. Introduction of a male to such a group promptly induced breeding readiness in the females, as indicated by increased gonadal hormone production and cyclic variation in estrogen and progesterone levels. The contrasting responses of group-living or single females to a new male suggest that at least two processes are involved in the induction of breeding readiness by group formation in females. One is a "priming" effect resulting from interfemale stimulation: the other is a "triggering" effect resulting from the presence of a male.

Contrasting responses to intruders and to involuntary separation by monogamous and polygynous New World monkeys.

The strength and quality of the relationship between heterosexual pairmates were compared in two species of New World monkeys, the polygynous squirrel monkey (Saimiri) and the monogamous titi monkey (Callicebus). This was accomplished by measuring behavioral and adrenocortical responses to intruders and to involuntary separation from the pairmate. The species diverged sharply in their responses to these conditions. For titi monkeys, the presence of an intruder resulted in a marked increase in behavioral signs of agitation, which were more prominent in the subject of the same sex as the intruder. Most male titi monkeys showed an increase in plasma cortisol in the presence of a male intruder; this pattern was not seen in their response to a female intruder. Female titi monkeys showed no consistent pattern of adrenocortical response to intruders of either sex. For squirrel monkeys, the presence of an intruder resulted in increased frequency of displays in male subjects, but not in female subjects. Both sexes showed a reliable reduction in plasma cortisol, below basal levels, in the presence of a female intruder, whereas no effect on cortisol was produced by a male intruder. Titi monkeys, but not squirrel monkeys, responded to separation with a significant increase in plasma cortisol and in behavioral indices of agitation or distress.

Behavioral and endocrine consequences of heterosexual pair formation in squirrel monkeys.

The South American squirrel monkey (Saimiri sciureus) typically lives in large social groups containing several individuals of all age/sex categories. When living in established heterosexual pairs, reproduction in this seasonally breeding primate is poor. We attempted to induce breeding activity in pair-housed monkeys by forming new heterosexual pairs just prior to the breeding season. Breeding readiness, as reflected in behavior and gonadal hormones, was induced in males, but not in females. Males also showed persistent increases in cortisol levels following formation of new heterosexual pairs; females did not. The results indicate that social stimulation provided by a single novel female is sufficient to enhance breeding readiness in male squirrel monkeys. Females, on the other hand, are apparently unresponsive to a single male whether novel or familiar; this may account for the poor reproductive success in squirrel monkeys housed in heterosexual pairs.

Sociophysiology of relationships in squirrel monkeys. I. Formation of female dyads.

Physiological and behavioral consequences of formation of female-female relationships were investigated in adult squirrel monkeys. Plasma cortisol, heart rate, and behavior were evaluated during confinement in a test cage while animals were housed alone, during the first hour of isosexual pair formation, and while animals were housed as isosexual dyads. In addition, basal cortisol levels were assessed both before and after pair formation, as were behavioral and cardiovascular responses to social and nonsocial stimulus presentations. Basal cortisol levels underwent a marked and sustained reduction following formation of pairs, independent of both dominance status and the quality of social interactions between pairmates. In contrast, adrenocortical responsiveness was not altered by social conditions, apart from a modest and transitory difference between dominant and subordinate females in their initial response to pair formation. Heart rate declined sharply during each test session, but did not reliably differentiate social conditions, stimulus conditions, or dominance status. These findings contrast with results of a parallel study of male squirrel monkeys and suggest that isosexual relationships in males and females are associated with different sociophysiological processes.

Behavioral and physiological adaptation to repeated chair restraint in rhesus macaques.

Physical restraint is a commonly used procedure when working closely with nonhuman primates. Nonhuman primates show rapid behavioral changes when learning the restraint procedure, and these changes have been taken to reflect behavioral and physiological habituation to the procedure. This study examined the behavioral and adrenocortical responses to repeated physical restraint in a large sample of adult male rhesus monkeys. Subjects showed a decline in behavioral agitation and cortisol concentrations across seven consecutive days of restraint. The changes in adrenocortical responsiveness were also coincident with an increased sensitivity to dexamethasone and a change in early morning basal cortisol secretion. The subjects were restrained for a single session 6 months later, and while the reduction in behavioral agitation was still present, the majority of changes in adrenocortical responsiveness were no longer present. These data show that behavior is not necessarily an indicator of underlying physiological processes and that the reduction of hypothalamic-pituitary-adrenal (HPA) activity with repeated restraint is due to physiological adaptation to high glucocorticoid concentrations and not to psychological habituation to the restraint procedures.

Endocrine sensitivity to novelty in squirrel monkeys and titi monkeys: species differences in characteristic modes of responding to the environment.

The present study examined plasma cortisol and behavioral responses to environmental novelty in squirrel monkey and titi monkey male-female pairs. Overall, seemingly trivial increments in novelty evoked sustained plasma cortisol elevations. In individually tested animals, the minimal level of novelty sufficient to evoke a cortisol response was smaller, and the ability of the response to discriminate among levels of novelty was greater, in titis than in squirrel monkeys. When tested with the pairmate, the sensitivity of the response was reduced in titis but not in squirrel monkeys. Behavioral measures were not as sensitive to novelty as was the cortisol response. The results suggest that differential endocrine responsiveness to novelty is an important physiological concomitant to previously described differences between squirrel monkeys and titi monkeys in their characteristic modes of relating to the environment.

Seasonality in squirrel monkeys (Saimiri sciureus): social facilitation by females.

The extent to which social living arrangements influenced seasonal changes in physiology and behavior was examined in adult squirrel monkeys (Saimiri sciureus). Data were collected over 20 months (encompassing two breeding seasons) from animals that were housed in three mixed-sex social configurations that varied in the number of heterosexual and isosexual social partners. For both sexes, the presence of multiple females was found to facilitate reproduction. Females housed with other females were more likely to exhibit seasonal ovarian cyclicity and tended to have higher conception rates. Social facilitation of reproduction was particularly prominent for subordinate females. The presence of same-sex companions also resulted in reduced adrenocortical output in females during the first nonbreeding season. In males, the availability of multiple females increased plasma testosterone levels, except in the presence of more dominant males. Seasonal increases in male weight coincided with increased cortisol levels and were most prominent in social groups containing multiple females. The influence of multiple females on male seasonality occurred despite the finding that male-female interactions were infrequent and, in fact, occurred less frequently when isosexual partners were available. Unexpectedly, affiliative social interactions between same-sex and opposite-sex partners occurred less frequently during the breeding season. As expected, behaviors associated with sex tended to increase during the breeding season. Very little agonism was observed during the course of the study and there was no evidence of interanimal competition for mates.

Sexual segregation in squirrel monkeys (Saimiri sciureus): a transactional analysis of adult social dynamics.

Female agonism against males and male interventions in social transactions between the sexes are 2 explanations for the low rates of social engagement observed between male and female squirrel monkeys (Saimiri sciureus). These possibilities were evaluated by comparing the frequency, structure, and content of transactions between the sexes in 3 experimental social units: male-female pairs, single-male-multifemale groups, and multimale-multifemale groups. Affiliative transactions between the sexes occurred 3-5 times less often in multimale-multifemale groups than in male-female pairs and in single-male-multifemale groups. Intermale agonism in the multimale-multifemale groups often coincided with ongoing transactions between the sexes, whereas female agonism against males was rather uncommon in all social units. The results failed to support the female agonism hypothesis and indicate a need for more detailed studies of intermale social dynamics.

Differences between Saimiri sciureus and Callicebus moloch in physiological responsiveness: implications for behavior.

The New World primates Callicebus moloch and Saimiri sciureus differ markedly in life modes. Physiological responsiveness (heart rate, cortisol) of these related taxa was compared in two situations that differed in the presumed degree of stress imposed, exposure to a novel environment and physical restraint. In both situations, Saimiri exhibited higher initial levels of heart rate and corticosteroids. In addition, the species differed in their patterns of response to both situations. Saimiri maintained initial heart rate levels across a 1-hour exposure period, whereas Callicebus showed a marked and continuous decline in heart rate. Adrenocortical activity increased in both species in response to test situations, but the elevations in plasma corticosteroids were substantially greater for Saimiri than for Callicebus. The results indicate that the species differ with respect to organization of physiological regulatory systems in a manner consistent with behavioral contrasts between them.

Comparative studies of social behavior in Callicebus and Saimiri: Behavioral and physiological responses of established pairs to unfamiliar pairs.

Comparisons were made of the behavioral and physiological responses of heterosexual pairs of two closely related New World primates, Callicebus moloch and Saimiri sciureus, to confrontations with unfamiliar pairs. Subject pairs were exposed to strangers in two situations differing in the presumed degree of arousal they imposed: obligatory (full visual exposure) and facultative (one part of the test cage allowed visual withdrawal). Behavioral reactions were evaluated employing frequencies of affiliative behaviors, agonistic behaviors, and spatial distribution. Physiological reactions were evaluated employing heartrate and plasma corticosteroid levels. Callicebus males and females were similarly affected by unfamiliar pairs. Both sexes behaved agonistically towards strangers; pairmates drew closer together and behaved in a coordinated fashion. Display behaviors declined, and spatial withdrawal increased over time. Heartrate in both sexes increased during confrontations, particularly during obligatory exposure. The reactions of Saimiri to strangers differed between the sexes. Males reacted agonistically toward unfamiliar conspecifics, as reflected in displays and spatial adjustments. In contrast, females showed a strong attraction to unfamiliar females. Despite the sex differences in behavior, heartrate did not differentiate confrontation conditions clearly in either sex. Plasma cortisol measures failed to differentiate between experimental conditions in either species.

Long-term exposure to intranasal oxytocin in a mouse autism model.

Oxytocin (OT) is a neuropeptide involved in mammalian social behavior. It is currently in clinical trials for the treatment of autism spectrum disorder (ASD). Previous studies in healthy rodents (prairie voles and C57BL/6J mice) have shown that there may be detrimental effects of long-term intranasal administration, raising the questions about safety and efficacy. To investigate the effects of OT on the aspects of ASD phenotype, we conducted the first study of chronic intranasal OT in a well-validated mouse model of autism, the BTBR T+ Itpr3tf/J inbred strain (BTBR), which displays low sociability and high repetitive behaviors. BTBR and C57BL/6J (B6) mice (N=94) were administered 0.8 IU/kg of OT intranasally, daily for 30 days, starting on day 21. We ran a well-characterized set of behavioral tasks relevant to diagnostic and associated symptoms of autism, including juvenile reciprocal social interactions, three-chambered social approach, open-field exploratory activity, repetitive self-grooming and fear-conditioned learning and memory, some during and some post treatment. Intranasal OT did not improve autism-relevant behaviors in BTBR, except for female sniffing in the three-chambered social interaction test. Male saline-treated BTBR mice showed increased interest in a novel mouse, both in chamber time and sniffing time, whereas OT-treated male BTBR mice showed a preference for the novel mouse in sniffing time only. No deleterious effects of OT were detected in either B6 or BTBR mice, except possibly for the lack of a preference for the novel mouse's chamber in OT-treated male BTBR mice. These results highlight the complexity inherent in understanding the effects of OT on behavior. Future investigations of chronic intranasal OT should include a wider dose range and early developmental time points in both healthy rodents and ASD models to affirm the efficacy and safety of OT.