Betty Zimmerberg

Glue sniffing is epidemic among children living in poverty in Latin America. Previous research has shown that abused inhalants such as toluene share pharmacological properties with anxiolytic drugs, and that personality factors such as degree of anxiety have been proposed to modulate the effects of these drugs. To study this interaction in an animal model, rats selectively bred for high (High) or low (Low) rates of distress calls after maternal separation (ultrasonic vocalizations, USVs) were used to investigate toluene's acute and long-term effects on two measures of anxiety behavior. At ten days of age, neonatal subjects were administered toluene (1 g/kg i.p.) and USVs were recorded. The subjects were retested as juveniles on an elevated plus maze to examine sequela of earlier toluene exposure. Acute toluene administration reduced USVs relative to control groups in neonates of both lines, indicating anxiolysis. As expected, Lows had reduced USVs relative to Highs. At 28 days of age, Highs spent more time in the open arms of the elevated plus maze than Lows. However, prior neonatal toluene exposure blocked this reversal of behavioral phenotype. This suggests that early toluene exposure compromised a compensatory process occurring during this developmental period, which may have been maternally mediated. These results have implications for the effects of early drug exposure on plasticity in the developing nervous system.

The effects of prenatal exposure to alcohol on behavioral correlates of catecholaminergic function were examined in three experiments. In each experiment, subjects had one of 3 prenatal treatment histories: prenatal alcohol-exposed (35% ethanol-derived-calories, 35% EDC), nutritional ...

Early childhood deprivation is associated with an increased risk of attachment disorders and psychopathology. The neural consequences of exposure to stress early in life have used two major rodent models to provide important tools for translational research. Although both models have been termed maternal separation (MS), the paradigms differ in ways that clearly shift the focus of stress between maternal and offspring units. The first model, here called early deprivation (ED), isolates pups individually while the dam is left not alone, but with a subset of littermates in the home nest ("stay-at-homes"). The other model, here called MS, isolates the dam in a novel cage while the pups are separated together. In this study, these two early stress models were directly compared for their effects on social behaviors in male and female juvenile offspring. Although both models altered play behavior compared to controls, patterns of prosocial behaviors versus submissive behaviors d...

Some of the behavioral deficits caused by prenatal or postnatal alcohol exposure have been demonstrated to be ameliorated by environmental manipulations such as handling or environmental enrichment. This experiment, in contrast, investigated whether behavioral deficits due to prenatal alcohol exposure could be exacerbated by a stressful experience, early weaning. Pregnant dams were given either a liquid diet with 35% of the calories derived from alcohol, a liquid diet without alcohol to control for any effects of the liquid diet administration, or ad libitum food and water. Half of each litter were weaned at 15 days of age (early weaning) and half were weaned at 21 days of age (normally weaned). Offspring were weighed, tested for activity in an open field at 18 days of age, and trained to find a hidden platform in the Morris water maze at 22-24 days of age. Alcohol-exposed subjects who were weaned early were more impaired in spatial navigation ability than any other group. Similarly, the combination of early weaning and prenatal alcohol exposure caused the slowest growth. All subjects exposed to alcohol, regardless of weaning condition, had greater latencies to find the platform than those from the two control groups. There was no synergistic effect of alcohol and stress on activity levels, but all early-weaned females were more active than normally weaned females; males did not show this effect. Thus, environmental stressors such as early weaning can compound detrimental symptoms of prenatal alcohol exposure. These results have implications for the understanding of the effects of the environment on neuronal plasticity.

Stress early in life exerts persistent detrimental effects on brain development. In this experiment, a rodent model of child neglect, early deprivation (ED), was used to investigate the role of the neurosteroid allopregnanolone [AlloP; 3alpha-hydroxy-5alpha-pregnan-20-one (3alpha,5alpha-THP)] in the development of anxiety behavior. Subjects were either undisturbed controls or ED: separated individually for 6 h per day from postnatal day (PN) 2 to 6. Control and ED subjects were also either noninjected, vehicle-injected or injected with 5 mg/kg AlloP prior to the isolation. At PN 7, responses to 2.5 or 5 microg icv injections of AlloP were determined for separation-induced ultrasonic vocalizations (USVs). Tolerance to the USV-reducing effect of daily AlloP was seen in control but not ED pups, and daily AlloP reversed the expected ED suppression of USVs. As adults, controls treated with postnatal AlloP were less anxious than all other groups on the elevated plus maze. ED counteracted this effect. Male controls showed a reversal of the typical sex difference. There were no effects on open-field activity. These results suggest that the neonatal brain is responsive to alterations in AlloP levels, and that neuroactive progesterone metabolites may play a role in mediating the development of stress-related sex differences.

... Thus, each yoked pair received the same relative volumeof diet (ml/kg) and hence the same number of calories on a bodyweight basis ... 3-44.34 Valentino, KL and Jones, EG, The early formation of thecorpus callosum: a light and electron microscopic study in foetaland neonatal ...

Allopregnanolone (3 alpha-hydroxy-5 alpha-regnan-20-one) is a neuroactive steroid recently shown to be involved in the neurochemical stress response via its positive modulation of the GABAA receptor complex. This experiment investigated the effects of postnatal stress (daily maternal separation during the first week of life) on the subsequent adult response to a stressor (10 min forced swim) in Long-Evans rats from one of three prenatal treatment groups (alcohol, pair-fed and control). Indices of stress response were allopregnanolone concentrations in plasma, cortex and hippocampus, and dopamine and norepinephrine concentrations in prefrontal cortex, nucleus accumbens and striatum. Females had higher levels of allopregnanolone than males in both plasma and brain. Prenatal alcohol exposure combined with early maternal separation stress resulted in an increase in the endogenous levels of allopregnanolone in the prefrontal cortex and hippocampus of adult offspring in response to a stressor compared to subjects without a prior history of postnatal stress; this effect was greater in females. This increased allopregnanolone was also associated with decreased dopamine and norepinephrine levels in the prefrontal cortex. In the prenatal alcohol-exposed offspring, postnatal maternal separation blunted the increase in dopamine levels in the striatum seen in both control groups. Postnatal maternal separation increased norepinephrine levels in the nucleus accumbens regardless of prenatal experience, while in the prefrontal cortex only prenatal diet condition (pair-feeding and alcohol) resulted in lower norepinephrine levels. The results of this experiment suggest that experience, both pre- and postnatal, can have long-term consequences for the developing neurochemical responses to stressors.

These experiments examined the role of gonadal hormones at both the organizational and activational time periods on sex differences in plus-maze behavior. In the first experiment, adult female Long-Evans rats were found to spend more time on the open arms of the plus maze than adult males, indicating less anxious behavior. In the second experiment, male and female subjects received a neonatal treatment (chemical castration with flutamide or tamoxifen, vehicle injection, or no injection) and a prepubertal treatment (gonadectomy, sham surgery, or no surgery). Adult females receiving either neonatal tamoxifen or prepubertal ovariectomy spent less time on the open arms than control females, but females who received both treatments were the most defeminized subjects. Males were not affected by the absence of gonadal hormones at either time period. These experiment indicate that female gonadal hormones play an important role both organizationally and activationally in plus-maze behavior. The role of the GABA receptor complex in mediating this effect is discussed. Knowledge of sex differences in plus-maze behavior may help to make this maze a more useful tool in investigating anxiety behavior in rats.

Although recent research has focused on the behavioral teratology of cocaine, there has been little attention centered on the effects of cocaine use on parenting behaviors after birth. This experiment investigated the effects of cocaine on parental (generally termed maternal) behavior in the rat. Four types of adult rats were tested for these effects over a 10-day testing period: lactating primiparous female dams, primiparous females, nulliparous females, and males. Subjects were injected daily with cocaine (5 mg/kg) or saline. Ten minutes postinjection each subject was placed in a cage with three stimulus pups and nesting material, and observed for several measures of maternal behavior for 5 minutes, including sniffing, licking, and retrieving pups. Subjects were also rated on a maternal behavior scale at 60 minutes and 24 hours postinjection, and on nest quality. Cocaine reduced both the total number and duration of sniffing and licking maternal behaviors, and resulted in a lower maternal behavior rating at 60 minutes compared to saline-treated subjects. Among those subjects that retrieved pups within the first 5 minutes, cocaine-injected subjects retrieved for a longer duration. These results also suggest impaired maternal behavior, since rapid retrievals would have been more efficient in maintaining a nest huddle. Thus, cocaine impaired the parenting ability of both male and female rats, with and without previous parenting experience.

The effects of prenatal alcohol exposure on the behavioral response to the neuroactive steroid allopregnanolone (3 alpha-hydroxy-5 alpha-pregnan-20-one) were investigated in neonatal rats. Two behaviors were assessed: retention of an odor conditioning task and production of ultrasonic vocalizations after brief maternal separation. Subjects from one of the three prenatal conditions (lab chow, alcohol, or pair-fed) received either no injection or an ICV injection of vehicle or one of three doses (1.25-5.0 micrograms) of allopregnanolone either 20 min prior to or immediately after training in an appetitive odor association paradigm. Retention was assessed 1 h later in a two-choice odor preference chamber. Posttraining injections of allopregnanolone caused a dose-dependent impairment in retention in the odor task, but there was no differential sensitivity to allopregnanolone in the alcohol-exposed offspring. All pretraining injections, including the vehicle, resulted in impairments in retention on the task, suggesting an impairment due to stress but not due to allopregnanolone. Allopregnanolone also reduced ultrasonic vocalizations after brief maternal separation in all subjects in a second experiment, but alcohol-exposed offspring displayed a dose-dependent shift to the right in their anxiolytic response to this neurosteroid. This decreased sensitivity suggests that prenatal alcohol exposure may cause a decrease in the density or affinity of the GABA receptors involved in stress response, but not cognitive processes, at this age.

The development of emotional behavior is dependent on the early experiences of the infant and the quality of maternal care. In these experiments, the effects of social isolation during the preweaning period on both pup behavior and maternal responsivity were examined. In the first study, the number of ultrasonic vocalizations (USVs) emitted after brief maternal separation was measured in neonatal rats with differing histories of social isolation. The social isolation procedure consisted of 5 days of daily separation from the dam and littermates for either 3 or 6 hr. At both ages tested, socially isolated pups vocalized significantly less than control pups. In the second study, the effects of prior isolation either daily for 5 previous days (Chronic Isolation) or for 4 hr prior to testing (Acute Isolation) were examined in a T-maze choice test. Pup vocalizations in the presence of the dam and dams' maternal behavior were assessed. When the dam was confined to the start box or during the maternal free access period, both Chronic and Acute Isolates vocalized less than pups that had never left the home nest. Dams spent more time with and licked and groomed more frequently and for a longer time both Chronic and Acute Isolates compared to pups that had always been with dams in the home nest. These results suggest that early isolation experience can alter subsequent responses to separation stress in neonatal rats and that maternal behavior is sensitive to the prior experiences of offspring.

Allopregnanolone, 3 alpha-hydroxy-5 alpha-pregnan-20-one (3 alpha,5 alpha-THP), a progesterone metabolite, is an endogenous neurosteroid mediating affective behaviors via its positive modulation of GABA(A) receptors. In order to better understand the role of this neurosteroid in individual differences in affective behavior, we used an animal model based on selective breeding for an infantile affective trait, ultrasonic vocalizations (USV). Adult male and female (in either proestrus or diestrus) rats that had been bred for low (low line) or high (high line) rates of USV after maternal separation were tested in a series of affective behavioral tests: open field, emergence, social interaction, defensive freezing, and the Porsolt forced swim task. Concentrations of allopregnanolone in combined hippocampus and amygdala tissue were then measured. low line subjects showed significantly lower anxiety and depression responses in the emergence, open field, and Porsolt forced swim tasks than did high line subjects. Proestrus females exhibited less affective behaviors than diestrus females or males. Allopregnanolone levels in hippocampus/amygdala were significantly higher in low line subjects compared to high line subjects, and in proestrus females compared to diestrus females and males. These data indicate that: (1) affective behaviors in lines selectively bred for an infantile anxiety trait exhibit selection persistence into adulthood; and (2) levels of allopregnanolone in the limbic system parallel selected disparities in affective behavior, suggesting a selection for alterations in the neurosteroid/GABA(A) receptor system in these lines.

The effects of prenatal alcohol exposure on learning in adult offspring were studied in a spatial task in a T-maze. Male and female Long-Evans rats were selected from litters whose dams had received one of three treatments: alcohol in a liquid diet (35% ethanol-derived calories, 35% EDC), pair-fed nutritional control (0% ethanol-derived calories, 0% EDC) or standard control (lab chow, LC). The task included trial-independent (reference memory) and trial-dependent (working memory) components: subjects were required to make a fixed left-right discrimination, and then to alternate left and right choices to escape water. Prenatal alcohol exposure was associated with a greater number of reference errors for both sexes; only males from the alcohol prenatal treatment group, however, were impaired on the working memory component. These results suggest that prenatal exposure to alcohol can cause behavioral dysfunctions that persist into adulthood. Secondly, the pattern of memory impairments suggests that both sexes may be equivalently damaged in neural areas subserving reference memory, but that males are selectively more vulnerable in neural areas subserving working memory.

Despite extensive research on the relationship between acute stress and hippocampal function in adults, little is known about the short- and long-term effects of prolonged juvenile stress on learning, memory, and other hippocampal functions. This experiment investigated whether spatial learning would be altered in juvenile and adult rats previously exposed to a chronic stressor: 6 h of social isolation (SI) daily at 15-21 days of age. SI was found to increase circulating plasma levels of corticosterone (CORT) and allopregnanolone (3-alpha,5-alpha-pregnan-20-one; 3,5-THP) at 1 h after separation on the fourth day, indicating that the isolation was an effective stressor. When tested as juveniles (post-natal (PN) 22-24), spatial learning was impaired on the Morris water maze in the previously isolated subjects compared to non-isolated controls. However, when tested as adults (PN 92-94), subjects previously exposed to SI during the third week of life demonstrated more rapid learning of the task than controls. These results are discussed in light of research on the effects of CORT on the developing hippocampus.

Duration and intensity of lordosis is mediated by actions of the progesterone (P) metabolite, 3 alpha-hydroxy-5 alpha-pregnan-20-one (3 alpha,5 alpha-THP) at GABA-sub(A) receptors in the midbrain ventral tegmental area. Because rats selectively bred for infantile vocalizations may differ in endogenous 3 alpha,5 alpha-THP secretion, their sexual behavior, midbrain, and plasma 3 alpha,5 alpha-THP levels as adult rats in behavioral estrus was examined. Rats bred for high rates of infantile vocalizations had shorter latencies and intervals between intromissions and ejaculation, higher lordosis quotients and ratings, more pacing of their sexual contacts, and had higher P and 3 alpha,5 alpha-THP levels in plasma and midbrain than did rats bred for low rats of infantile vocalizations. Thus, levels of 3 alpha,5 alpha-THP in the midbrain are associated with differences in sexual behavior of these rats.

Animal models of child neglect (known as maternal separation or early deprivation) have suggested a causal link to subsequent depression and/or anxiety in children. In this experiment, the acoustical features of the ultrasonic calls emitted by a rat pup when separated from its dam were analyzed as well as the maternal behavior when the dam was allowed to retrieve the pup. Bout structure and harmonic double shifts did differ between controls and "neglected" pups, as did maternal attention. This model will be used to determine neural mechanisms underlying deficits in attachment behavior.