Non-handled Research Articles

Long maternal separation has protective effects in rats exposed to activity‐based anorexia

This study examined the effect of three neonatal treatments of maternal separation during infancy in young adult rats exposed to standard activity-based anorexia (ABA) consisting of food restriction plus free access to an activity wheel. During the first 20 postnatal days of life rat pups were exposed to periods of either brief maternal separation (BMS, 15 min), long maternal separation (LMS, 180 min), or were non-handled (NH). Thereafter, male and female rats were exposed to ABA. Neonatal treatment produced no significant differences in the survival time of male rats, whereas survival was greater in female rats exposed to LMS than in NH rats under ABA procedure. In conclusion, prolonged maternal separation appears to promote resistance in female animals subjected to harsh ABA life-threatening conditions.

Neonatal handling and the maternal odor preference in rat pups: Involvement of monoamines and cyclic AMP response element-binding protein pathway in the olfactory bulb

Early-life environmental events, such as the handling procedure, can induce long-lasting alterations upon several behavioral and neuroendocrine systems. However, the changes within the pups that could be causally related to the effects in adulthood are still poorly understood. In the present study, we analyzed the effects of neonatal handling on behavioral (maternal odor preference) and biochemical (cyclic AMP response element-binding protein (CREB) phosphorylation, noradrenaline (NA), and serotonin (5-HT) levels in the olfactory bulb (OB)) parameters in 7-day-old male and female rat pups. Repeated handling (RH) abolished preference for the maternal odor in female pups compared with nonhandled (NH) and the single-handled (SH) ones, while in RH males the preference was not different than NH and SH groups. In both male and female pups, RH decreased NA activity in the OB, but 5-HT activity increased only in males. Since preference for the maternal odor involves the synergic action of NA and 5-HT in the OB, the maintenance of the behavior in RH males could be related to the increased 5-HT activity, in spite of reduction in the NA activity in the OB. RH did not alter CREB phosphorylation in the OB of both male and females compared with NH pups. The repeated handling procedure can affect the behavior of rat pups in response to the maternal odor and biochemical parameters related to the olfactory learning mechanism. Sex differences were already detected in 7-day-old pups. Although the responsiveness of the hypothalamic–pituitary–adrenal axis to stressors is reduced in the neonatal period, environmental interventions may impact behavioral and biochemical mechanisms relevant to the animal at that early age.

Sustained hyperphagia in adolescent rats that experienced neonatal maternal separation

To examine the neurobiological basis of bingeing-related eating disorders using an animal model system. Sprague-Dawley pups were separated from dam for 3 h daily during the first two weeks of birth (maternal separation (MS)), or left undisturbed (non-handled (NH)). Pups were subjected to repeated fasting/refeeding (RF) cycles; that is, 24 h food deprivation and 24 h RF (NH/RF or MS/RF), or had free access to food and water (NH/fed control (FC) or MS/FC) from postnatal day (PND) 28-40. Body weight gain and food intake were recorded. The arcuate expression of neuropeptide Y (NPY) and plasma corticosterone levels were analyzed on PND 29 and 40. Decrease in weight gain by repeated fasting/RF cycles was smaller in MS pups than in NH. Interestingly, weight changes responding to fasting or RF increased in MS/RF compared with NH/RF. Compensatory hyperphagia was diminished in NH/RF after the third fasting trial, but persisted in MS/RF throughout the experimental period. The arcuate expression of NPY mRNA responding to food deprivation was blunted, but elevation of plasma corticosterone exaggerated, in the MS group, compared to the NH group, on PND 29 after the first fasting session. However, both the arcuate NPY mRNA and plasma corticosterone levels were increased in MS/RF, but not in NH/RF, on PND 40 after the six sets of fasting/RF cycles, compared to the free FC groups. Experience of neonatal MS may lead to an exaggerated feeding response to repeated fasting/RF challenges at adolescence, perhaps, due to increased responsiveness of the hypothalamic-pituitary-adrenal gland axis. Additionally, the results suggested that an increased action of the hypothalamic NPY may not be necessary to induce compensatory hyperphagia following food deprivation.

Frequency of maternal licking and grooming correlates negatively with vulnerability to cocaine and alcohol use in rats

Because licking and grooming behavior of dams with pups can influence some behaviors of pups when they are adults, we tested if licking and grooming scores in a maternal separation protocol correlated with cocaine or ethanol self-administration in the pups as adults. The protocol produced litters that were separated from dams for 0 (MS0), 15 (MS15) or 180 (MS180) min, and a nonhandled (NH) group as well. Self-administration of both drugs as shown in earlier studies was lowest in the MS15 group, highest in the NH group and intermediate in the other groups. Licking and grooming scores correlated negatively with drug intake and suggests that maternal care of pups can influence drug use in pups when they are adults.

Early Postnatal Stress Alters Place Conditioning to Both μ- and κ-Opioid Agonists

Clinical literature has established a link between early childhood incidents of neglect and trauma and adult problems with substance abuse. In rats, such early life stress has been modeled using a maternal separation (MS) paradigm in which rat pups were removed from their mothers for a few hours daily during the first two postnatal weeks. In this study, we used the MS model to investigate the effects of early postnatal stress on place conditioning to both mu- and kappa-opioid agonists in male and female Long-Evans rats. Offspring of both rearing conditions [MS or nonhandled (NH)] were conditioned using a biased procedure to saline, the mu-opioid agonist morphine (3.0, 5.6, and 10 mg/kg s.c.), or the kappa-opioid agonist spiradoline (0.3, 1.0, and 3.0 mg/kg) for 3 days, followed by a drug-free place-conditioning test 24 h later. Saline was administered in the morning, 30 min before confinement in one compartment, whereas morphine or spiradoline was administered in a similar manner 6 h later in the opposite compartment. MS offspring spent significantly more time in the morphine-paired compartment than NH offspring, indicating a greater place preference for the mu-opioid agonist. In the case of spiradoline, NH offspring spent significantly less time in the spiradoline-paired compartment, indicating a greater aversion to the kappa-opioid agonist in these animals than in MS offspring. These findings indicate that early postnatal stress can significantly alter the rewarding or aversive value of mu- and kappa-opioid agonists when measured using place conditioning.

Maternal Separation Leads to Persistent Reductions in Pain Sensitivity in Female Rats

We determined responses to noxious thermal stimuli, before and after morphine, and mu-opioid receptor binding in brain regions involved in nociception in maternally separated (MS), neonatally handled (H) and nonhandled (NH) female rats. Long-Evans dams were randomly assigned to either 180-minute (MS) or 15-minute (H) minute daily separations from their litters or left undisturbed (NH). At 120 days of age, paw lick latency (50 degrees C hot plate) was determined in offspring during diestrous. Rats were then given 1, 2, 5, or 10 mg/kg morphine and paw lick latency was measured. Rats were killed during diestrous and mu-opioid receptor binding was determined in discrete brain regions, using [(3)H]DAMGO autoradiography. MS rats had significantly longer (P < .05) paw lick latencies compared with H rats. The percent maximal possible effect of morphine was significantly (P < .05) lower in MS compared with H rats for the 5 mg/kg dose. Mu-Opioid receptor binding capacity was significantly greater (P < .05) in MS rats compared with H rats in the medial preoptic nucleus. In conclusion, MS and H treatments led to antipodal differences in pain sensitivity in female rats and differential mu-opioid receptor binding in the medial preoptic nucleus. This article describes the persistent impact of early life adversity on pain sensitivity and the analgesic potency of morphine. Clinically, early life history may play an important role in pain symptoms and responses to opioid analgesics.

Enhanced sensitivity to naltrexone-induced drinking suppression of fluid intake and sucrose consumption in maternally separated rats

Early-life stress has been identified as a risk factor in the development of a host of disorders, including substance abuse; however the link between early postnatal stress and changes in measures of reward has not been thoroughly researched. The current study had two main objectives: 1) to determine the impact of maternal separation (an animal model of early-life stress) on the consumption of 10% and 2.5% sucrose solutions by Long–Evans rat dams and male and female offspring, and 2) to determine the effect of the opioid antagonist naltrexone (0.1–3.0 mg/kg) on drinking by each of those groups. Dam-pup separations occurred for varying lengths of time during the first two postnatal weeks. In Experiment 1, a two-bottle choice test (sucrose solution vs. water) was administered across five days to both nonhandled (NH) and maternally-separated (MS) offspring as adults and to dams 2–4 weeks post-weaning. In Experiment 2, naltrexone was administered prior to two-bottle choice tests. MS males and the dams of MS litters exhibited increased intake of total fluid and sucrose solutions, whereas results from females were less consistent. Naltrexone elicited a greater decrease in fluid intake and sucrose intake in male MS offspring compared to male NH offspring. These results indicate that early postnatal stress alters both sucrose consumption, a non-drug measure of reward, and apparently the brain opioid systems that mediate naltrexone-induced drinking suppression.

Age, sex and early environment contribute to individual differences in nicotine/acetaldehyde-induced behavioral and endocrine responses in rats

Neonatal handling was used to evaluate the influence of early environment on responses to nicotine. Rats exposed as pups to daily short-term separation from the dam (H) were compared to non-handled (NH) controls. In experiment 1, prepubescent males and females, aged postnatal day (P) 30, were tested for the effect of nicotine/acetaldehyde (NicAc) on open field behavior and plasma corticosterone levels. NicAc induced increases in ambulatory activity and time spent in the center of the field in NH, but not H, males. Drug-induced increases in initial ambulatory activity, but not center time, were also seen in NH and H females. Handling, but not sex, contributed to group differences in plasma corticosterone levels. In experiment 2, NH and H rats were tested for acquisition of NicAc self-administration at three ages, P27–31, P34–38 and P90–94. Age and sex, but not handling, contributed to differences in performance of this task. Whereas males exhibited a decrease in responding with age, females did not. These findings demonstrate that neonatal handling may serve as an experimental model for individual differences in sensitivity to tobacco constituents. Furthermore, the current study indicates that stress reactivity, age and sex may play differential roles in initiating smoking behavior.

Early handling reduces vulnerability of rats to activity‐based anorexia

Resistance to restricted feeding with and without wheel access was tested in rats handled (H) for 20 days since birth. Weight loss produced by 1.5-hr restricted food access was less in H than in non-handled (NH) males when tested aged 41 days. At this age combining food restriction with access to a running wheel (a procedure commonly known as activity-based anorexia, ABA) produced very rapid weight loss and no effect of handling was detected. When 75-day females were tested in the same way, under the ABA procedure H rats took longer than NH controls to reach the removal criterion. Simply restricting food access in these females produced variable weight loss, without detection of any handling effect. No differences in food intake or running were detected between H and NH rats in either males or females. In conclusion, handling seems to have a direct effect on rats' later response to either food deprivation alone or to an ABA procedure.

Long-lasting changes in behavioural and neuroendocrine indices in the rat following neonatal maternal separation: Gender-dependent effects

Neonatal maternal separation (MS) has been used to model long-term changes in neurochemistry and behaviour associated with exposure to early-life stress. This study characterises changes in behavioural and neuroendocrine parameters following MS. On postnatal days (PND) 3–15, male and female Long–Evans rats underwent 3 h daily MS. Non-handled (NH) control offspring remained with the dams. Starting at PND 90, behaviour was assessed at weekly intervals in the elevated plus-maze, elevated T-maze, and locomotor activity boxes, and body weight monitored throughout. At the end of the study, adrenals were weighed and blood collected for analysis of plasma corticosterone and adrenocorticotropic hormone (ACTH) under basal conditions and following restraint stress. As adults, MS weighed more than NH animals. Activity on the open arms of the plus-maze was similar between MS and NH animals. In the T-maze, MS males had shorter emergence latencies than their NH counterparts. Spontaneous ambulation in a novel environment was significantly higher in MS than in NH animals, and males exhibited overall lower activity than females. Basal plasma corticosterone was lower in MS than in NH females, but no rearing condition difference was observed following restraint stress. Females had higher corticosterone and ACTH levels than males, whereas adrenal glands of MS animals weighed less than those of NH controls. The MS paradigm caused long-term gender dependent effects on behaviour and HPA axis status. The consistent gender differences confirm and expand existing results showing altered anxiety and stress reactivity in male and female rats.

Mesocortical dopamine and HPA axis regulation: Role of laterality and early environment

The infralimbic (IL) cortex is importantly involved in regulating behavioral and physiological responses to stress, including those of the hypothalamic–pituitary–adrenal (HPA) axis. The mesocortical dopamine (DA) system is an important afferent modulator of this region, is highly stress sensitive and frequently shows functional hemispheric asymmetry. Postnatal handling stimulation facilitates development of cortical asymmetry and is also associated with optimal HPA axis regulation. The present study examines the poorly understood role of the mesocortical DA system in regulating HPA axis function in adult rats which were handled (H) or nonhandled (NH) postnatally. In the first experiment, unilateral intra-IL cortex injection of the DA (D1/D2) antagonist α-flupenthixol into either hemisphere significantly exaggerated the restraint stress-induced increases in plasma adrenocorticotrophic hormone and corticosterone in NH rats. In H rats, the same effect was lateralized to the right IL cortex. In a second experiment, post mortem neurochemical analysis of DAergic measures in the IL cortex was conducted in H and NH animals following either acute or repeated (5 times) restraint stress. DAergic measures in the right IL cortex were significantly correlated with reduced stress hormone activation in both H and NH rats, especially in repeatedly restrained rats. However, while H rats showed a significant rightward shift in DA metabolism with repeated stress experience, NH rats shifted DA metabolism to the left. It is suggested that, during stress, mesocortical DA release normally acts in an adaptive, negative feedback capacity preventing excessive HPA activation and, with repeated stress, the right IL cortex is particularly important in this capacity. As well, the selective enhancement of DA metabolism in the right IL cortex of H rats may underlie, in part, their typically superior ability to adapt to stress and constrain HPA activity.

Neonatal isolation alters estrous cycle effects on ventral striatal extracellular monoamine levels

We demonstrated that neonatal isolation (ISO) enhances cocaine self-administration in male and female adult rats and alters ventral striatal extracellular levels of serotonin (5-HT) and dopamine (DA) basally or in response to psychostimulants in infant rats. Now, we examine basal 5-HT, DA, and norepinephrine (NE) levels in nucleus accumbens (NAc) using in vivo microdialysis in adult male and female rats with or without ISO experience. NAc shows estrous cycle-dependent effects as do behavioral responses to cocaine. Because our prior work showed ISO eliminated estrous-cycle effects on behavior, we now test separate groups of females in proestrus, estrus, or diestrus stages. Litters were assigned to the ISO (1-h isolation; postnatal days 2–9) or non-handled (NH) condition. During adulthood (postnatal day 70–90), microdialysis probes were implanted and aimed at NAc core. Ten samples were collected over 150-min and measures of 5-HT, DA, and NE were analyzed via HPLC. ISO did not affect 5-HT levels in males. However, ISO modified estrous stage effects on 5-HT. The pattern of 5-HT levels in NH females (higher in diestrus and proestrus vs. estrus) was reversed in ISO females. DA levels were unaffected by ISO, similar to our findings at other ages, and did not differ by gender or estrous stage. None of these factors affected NE levels. Because 5-HT modulates DA and levels of both transmitters are increased by cocaine, this neurochemical effect of ISO may contribute to the ability of ISO to alter the behavioral responses to cocaine as we showed previously.

Fasting-induced increases of arcuate NPY mRNA and plasma corticosterone are blunted in the rat experienced neonatal maternal separation

This study was conducted to examine the effects of neonatal maternal separation on the hypothalamic expression of feeding peptides in later life. Pups in maternal separation (MS) groups were separated from their dam for 3 h daily from postnatal day (PND) 1-14, while pups in non-handled (NH) groups were left undisturbed. Rats were sacrificed on PND 60 to examine the gene expression of neuropeptide Y (NPY) and pro-opiomelanocortin (POMC) in the hypothalamic arcuate nucleus by mRNA in situ hybridization. Half of the rats from each group were food-deprived for 48 h before sacrifice. POMC mRNA expression increased in the free fed MS group compared with the free fed NH group. Food deprivation significantly decreased the arcuate POMC mRNA level in both groups. Body weight gain, basal levels of plasma corticosterone, leptin, and arcuate NPY mRNA were not modulated by experience of neonatal maternal separation. However, fasting-induced increases of plasma corticosterone and arcuate NPY expression were blunted in MS rats. These results suggest that neonatal maternal separation may increase the basal expression level of arcuate POMC mRNA, while inhibit the fasting-induced expression of arcuate NPY mRNA, later in life. Lastly, the altered expression of arcuate NPY mRNA, but not of arcuate POMC mRNA, appeared to be related with altered activity of the hypothalamic-pituitary-adrenal gland axis in offspring by neonatal maternal separation.

Neurochemical and Behavioral Responses to Cocaine in Adult Male Rats with Neonatal Isolation Experience

Our research demonstrates that neonatal isolation (ISO; 1 h/day isolation; postnatal days 2-9) enhances extracellular, ventral striatal dopamine (DA) responses to psychostimulants in infant and juvenile rats. In adult rats, we find ISO facilitates acquisition and maintenance of cocaine self-administration. We now test whether ISO enhances cocaine-induced accumbens DA levels in adults using in vivo microdialysis. Behavioral responses to cocaine and DA antagonists were also examined. Adult male rats were derived from litters subjected to ISO or nonhandled (NH) control conditions. In experiment 1, microdialysis probes were aimed at accumbens core and separate groups administered vehicle or cocaine (5 and 10 mg/kg i.p.). Samples were analyzed for DA levels via high-performance liquid chromatography. In experiment 2, ISO and NH rats were administered one of these cocaine doses, and locomotor activity was assessed. Effects of cocaine (0.3-30 mg/kg), the D(1) antagonist SCH23390 [R-(+)-7-chloro-8-hydroxy-3-methyl-1-phenyl-2,3,4,5-tetrahydro-1H-3-benzazepine (0.003-0.03 mg/kg)], and the D(2) antagonist eticlopride (0.01-0.1 mg/kg) on disruption of responding for food were examined in experiment 3. Cocaine plasma levels were assessed in experiment 4. ISO enhanced cocaine-induced increases in accumbens DA levels. Furthermore, the D(2), but not D(1), antagonist disrupted behavior to a greater extent in ISO versus NH rats. Yet, ISO did not significantly alter behavioral responses to cocaine or cocaine plasma levels. These data show that the ability of ISO to enhance accumbens DA responses to cocaine endures into adulthood. Moreover, that ISO rats are more sensitive to a D(2) antagonist may reflect decreased levels of this receptor type as we showed previously in infant rats.

Neonatal isolation alters the estrous cycle interactions on the acute behavioral effects of cocaine

We demonstrated that neonatal isolation (ISO) increases acquisition of cocaine self-administration and alters psychostimulant-induced ventral striatal dopamine and serotonin levels in female rats. Both dopamine and serotonin modulate the behavioral effects of cocaine and these effects can vary across estrous stages. We now test whether ISO modifies the manner in which estrous stage affects the acute behavioral responses to cocaine. Litters were assigned to ISO (1 h/day isolation; post-natal days 2-9) or non-handled (NH) conditions. In Experiment 1, the ability of cocaine (0.3-30 mg/kg; IP) to disrupt schedule-controlled responding for food was assessed in proestrus, estrus, and diestrus stages. Diestrus and proestrus NH females showed increased response rates at low cocaine doses and decreased rates at higher doses relative to baseline. In contrast, estrus NH females showed decreased responding across all doses. ISO eliminated this estrous stage distinction; only decreased responding to high cocaine doses were seen. Yet, estrous cyclicity during food restriction (Experiment 2) did not differ by group. To confirm this ISO effect, proestrus or estrus rats were administered cocaine (0, 5, 10 mg/kg; IP) and activity monitored in Experiment 3. Locomotor activity differed by estrous stage in NH but not ISO rats. Cocaine plasma levels (Experiment 4) at the time of peak behavioral activity did not differ by group or estrous stage. Results extend prior studies to show estrous stage alters the behavioral effects of cocaine. Neonatal isolation eliminates these effects perhaps reflecting alterations in accumbens monoamine levels or the effects of estrogen on this system.

Early social and physical deprivation leads to reduced social motivation in adulthood in Wistar rats

Behavioural abnormalities in adulthood may have their origin in a disturbed interaction with the environment during postnatal development. We tested the consequences for adult social motivation of early deprivation (ED) of rat pups from mothers and littermates relative to nonhandled (NH) pups. Early deprivation was performed at room or warm ambient temperatures, cold-ED and warm-ED, respectively, and during either the dark or light phase of the daily cycle. In adulthood, rats that were unrelated and unfamiliar but of the same treatment group were introduced in pairs to an open field for a 30-min test. Social behaviour in home base and exploration modes was assessed using algorithmic analysis of the XY locations of the two rats. Findings revealed that Cold-ED induced a preference for a separate home base, which limited significantly the episodes of social interactions, in comparison to NH. Warm-ED had no comparable effect on the rats’ social behaviour. These findings indicate that ED under ambient conditions that constitute severe thermal stress for rat pups leads to development of reduced social motivation in adulthood.

Neonatal isolation enhances acquisition of cocaine self-administration and food responding in female rats

We showed previously that neonatal isolation (ISO) enhances acquisition of cocaine self-administration in adult male rats without altering acquisition of food responding. Female rats show poorer performance in learning tasks and are differentially affected by stress compared to male rats. Thus, we investigated whether ISO alters acquisition of operant responding for cocaine and food in female rats with comparison to male rats. Litters were subjected to ISO or were non-handled (NH). Activity levels were assessed in adult rats. Then, rats were implanted with jugular catheters and allowed to self-administer cocaine under a fixed-ratio 1 (FR1) schedule of reinforcement using an escalating dose presentation procedure. Cocaine intake, discrimination of active versus inactive levers, and ineffective active lever responses were tabulated. Effects of non-contingent cocaine infusions (primes) and increasing FR on responding were then assessed. Other rats were allowed to lever press for food under an FR1 schedule (10 s time-out). ISO enhanced acquisition of operant responding for food and cocaine in female rats. The latter was demonstrated by better lever discrimination, emission of fewer ineffective responses, and superior performance in response to primes. Yet, NH females ingested more cocaine than ISO females during the initial acquisition phase. In male rats, ISO enhanced acquisition of cocaine self-administration but not food responding. Activity levels were unaffected by ISO or gender. These data confirm and extend our previous findings demonstrating the enduring effects of ISO on adult self-administration behavior and emphasize the importance of measuring behavioral patterns versus intake in acquisition studies.

Prenatal exposure to diazepam and alprazolam, but not to zolpidem, affects behavioural stress reactivity in handling-naı̈ve and handling-habituated adult male rat progeny

A gentle long-lasting handling produces persistent neurochemical and behavioural changes and attenuates the impairment in the behavioural reactivity to novelty induced by the prenatal exposure to diazepam (DZ) in adult male rat progeny. This study investigated the consequences of a late prenatal treatment with three GABA/BDZ R agonists (DZ) alprazolam (ALP) and zolpidem (ZOLP)), on different stress-related behavioural patterns, in non-handled (NH), short-lasting handled (SLH) and long-lasting handled (LLH) adult male rats exposed to forced swim test (FST), acoustic startle reflex (ASR) and Vogel test (VT). The effects on motor activity were evaluated in the open field and in the Skinner box. The seizure sensitivity to picrotoxin (PTX) was investigated as an index of the functional state of GABA/BDZ Rs. A single daily s.c. injection of DZ (1.25–2.50 mg/kg) and ALP (0.125–0.250 mg/kg) over gestational days 14–20 induced a decrease in immobility time in the FST in NH rats, no change in SLH rats and an increase in LLH rats; DZ induced an increase in the peak amplitude of the ASR in NH rats, no change in SLH rats and a reduction in LLH rats; ALP was ineffective in all groups. DZ and ALP reduced the number of punished licks in the VT in NH, SLH and LLH rats while the unpunished licks were not modified. DZ decreased locomotion and the lever pressing responses while ALP increased them. DZ and ALP increased the seizure sensitivity to PTX (2.5–4.0 mg/kg i.p.). These findings indicate a convergence on anxiety-related behaviours in the effects of prenatal exposure to DZ and ALP and a differentiation on motor activity. Long-lasting handling was able to overcompensate the increased behavioural stress reactivity induced by the prenatal exposure to DZ and ALP.

Prenatal ethanol exposure and spatial navigation: effects of postnatal handling and aging.

Prenatal ethanol exposure results in spatial navigation deficits in young and mid-aged animals. In contrast, postnatal handling attenuates spatial deficits that emerge with age in animals that are not handled. Therefore, we investigated the ability of handling to attenuate spatial deficits in animals prenatally exposed to ethanol (E). Sprague-Dawley male offspring from E, pair-fed (PF), and control (C) groups were handled (H) or nonhandled (NH) from 1 to 15 days of age and tested on the Morris water maze at 2 or 13 to 14 months of age. In young animals, H-E males had longer latencies to locate the submerged platform, and E animals, across handling conditions, showed altered search patterns compared to their PF and C counterparts. Mid-aged animals had longer latencies than young animals, with no differences among E, PF, and C animals. However, corticosterone levels were higher in mid-aged E than in C males. Handling did not attenuate impairments associated with either prenatal ethanol exposure or aging.

Neonatal isolation alters stress hormone and mesolimbic dopamine release in juvenile rats

Rat pups were individually isolated from the mother and nest for 1 h/day from postnatal days (PND) 2 to 9 and tested as juveniles (PND 26–30) compared to nonhandled (NH) controls. In response to 1 h of restraint stress, NH rats increased locomotor activity and dopamine (DA) levels, but neonatally isolated (ISO) rats did not. Both groups had increased plasma corticosterone levels in response to restraint, but corticosterone levels were higher in ISO than in NH. Brain allopregnanolone (3α,5α-THP) levels also increased in response to stress, but NH and ISO did not differ. Sex of the rats was not a factor for any of the measures except plasma corticosterone levels, where females had higher levels than males. These data indicate that the effects of neonatal isolation persist postweaning and that the effects are most evident in response to stress as opposed to under baseline conditions.