Role of medial prefrontal NMDA receptors in spatial delayed alternation in 19‐, 26‐, and 33‐day‐old rats

Long–Evans rats were trained on spatial delayed alteration (SDA) in a T‐maze following medial prefrontal cortical (mPFC) infusions of different doses of the noncompetitive NMDA‐receptor antagonist, MK‐801 (.125 µl; .25 µl; or .25 µlsaline, bilaterally), on postnatal day (PND) 19, 26, or 33. Pups trained on PND 19 showed almost no learning of SDA, regardless of drug condition (including saline). On PND 26, both doses of MK‐801 significantly and equivalently prevented SDA learning, with performance during the final three training blocks remaining near chance levels, in contrast with 85% correct performance in the saline control group. On PND 33, substantial SDA learning was evident regardless of dose, although a modest impairment appeared in mid‐training at both doses. These findings confirm previous reports of mPFC involvement in the early postnatal ontogeny of SDA and suggest a developmentally transient role of mPFC NMDA‐receptor function in this task. Dev. Psychobiol. © 2010 Wiley Periodicals, Inc. Dev Psychobiol 52: 583–591, 2010.     

In vitro binding assays using H nisoxetine and H WIN 35,428 reveal selective effects of gonadectomy and hormone replacement in adult male rats on norepinephrine but not dopamine transporter sites in the cerebral cortex

The prefrontal cortices mediate cognitive functions that critically depend on local dopamine levels. In male rats, many prefrontal tasks where performance is disrupted by changes in dopamine signaling are also impaired by gonadectomy, a manipulation that increases cortical dopamine concentration, prefrontal dopamine axon density and possibly extracellular prefrontal dopamine levels as well. Because these actions could be responsible for the impairing effects of gonadectomy on prefrontal function, the question of how they might arise comes to the fore. Accordingly, the present studies asked whether dopamine levels might be increased via a hormone sensitivity of transporter-mediated dopamine uptake. Specifically, ³H WIN 35,428 and ³H nisoxetine, ligands selective for the dopamine (DAT)- and norepinephrine transporter (NET) respectively, were used in in vitro binding assays to ask whether gonadectomy altered transporter affinity (Kd) and/or binding site number (Bmax) in prefrontal cortex, sensorimotor cortex and/or caudate. Assays performed on tissues dissected from sham-operated, gonadectomized and gonadectomized rats supplemented with testosterone propionate or estradiol for 4 or 28 days revealed no significant group differences or obvious trends in Kd or Bmax for DAT binding or in measures of Bmax for NET binding. However, affinity constants for ³H nisoxetine were found to be significantly higher in sensorimotor and/or prefrontal cortex of rats gonadectomized and gonadectomized and supplemented with estradiol for 4 or 28 days but similar to control in gonadectomized rats given testosterone. Because the NET contributes substantially to extracellular prefrontal dopamine clearance, these androgen-mediated effects could influence prefrontal dopamine levels and might thus be relevant for observed effects of gonadectomy on dopamine-dependent prefrontal behaviors. A hormone sensitivity of the NET could also have bearing on the prefrontal dopamine dysfunction seen in disorders like schizophrenia that disproportionately affect males, whose severity correlates with abnormal testosterone levels, and for which the NET is among suspected sites of pathology.     

Postnatal development of neurons expressing NADPH-diaphorase and parvalbumin in the parietal cortex of male and female rats

Expression of the enzyme nicotinamide adenine dinucleotide phosphate-diaphorase (NADPH-d) and the calcium-binding protein parvalbumin was studied in the parietal cortex of male and female rats during postnatal development at 20, 60 and 90 days of age. First, localization of the activity of NADPH-d was combined with the immunohistochemical localization of parvalbumin to facilitate recognition of morphological details and distribution patterns of these two types of cortical neurons. Double staining of neurons for parvalbumin and NADPH-d was never found. Second, it was found that NADPH-d is a simple and proper marker for quantitative studies. Morphometric analysis revealed sexual dimorphism in the density of NADPH-d-positive neurons in 20 days-old prepubertal rats. Females showed higher amounts of NADPH-d-positive neurons than males. No sex-dependent differences were detected in 60 days-old pubertal and 90 days-old postpubertal rats. The present results suggest that sex differences in the number of NADPH-d-positive neurons in the rat parietal cortex may be related to epigenetic effects of gonadal hormones in the early prepubertal period of postnatal development.     

Enhancing the salience of dullness: behavioral and pharmacological strategies to facilitate extinction of drug-cue associations in adolescent rats

Extinction of drug-seeking is an integral part of addiction treatment, and can profoundly reverse or ameliorate the harmful consequences of drug use. These consequences may be the most deleterious during adolescence. The studies presented here build from recent evidence that adolescent rats are more resistant to extinction training than adults, and therefore may require unique treatment strategies. We used unbiased place-conditioning in male rats to show that passive, un-explicit extinction pairings resulted in delayed extinction in 40-day-old adolescents relative to 80-day-old adults. However, explicit-pairing of a previously cocaine-associated context with the absence of drug produces extinction in adolescents as rapidly as in adults. These data suggest that successful extinction of drug-paired associations in adolescents may be facilitated by stronger acquisition of a new (extinction) memory. Drug-paired associations are largely controlled by the prelimbic prefrontal cortex (plPFC) and its influence on the nucleus accumbens (NAc). This pathway mediates the motivational salience attributed to incoming stimuli through the D1 dopamine receptor. D1 receptors on plPFC outputs to the accumbens are transiently overproduced during adolescence. Since D1 receptors are selectively responsive to potent stimuli, we hypothesized that the adolescent plPFC hinders competition between potent drug-paired associations and the subtler, drug-free information necessary for extinction. To harness this unique profile of the adolescent plPFC, we aimed to increase the salience of unrewarded extinction memories by activating plPFC D1 receptors during extinction training. In a second study, extinction of drug-cue associations was facilitated in adolescents by elevating dopamine and norepinephrine in the PFC during extinction training with atomoxetine. In a third study, direct microinjection of the D1 receptor agonist SKF38393 mimicked this effect, also facilitating extinction in adolescent subjects. Furthermore, pharmacological intervention attenuated subsequent drug-primed reinstatement of cocaine-conditioned preferences. We establish a potential direction for distinct strategies to treat this vulnerable population.     

Active versus latent representations: a neural network model of perseveration, dissociation, and decalage

Children of different ages often perseverate, repeating previous behaviors when they are no longer appropriate, despite appearing to know what they should be doing. Using neural network models, we explore an account of these phenomena based on a distinction between active memory (subserved by the prefrontal cortex) and latent memory (subserved by posterior cortex). The models demonstrate how (a) perseveration occurs when an active memory of currently relevant knowledge is insufficiently strong to overcome a latent bias established by previous experience, (b) apparent dissociations between children's knowledge and action may reflect differences in the amount of conflict between active and latent memories that children need to resolve in the tasks, and (c) differences in when children master formally similar tasks (decalage) may result from differences in the strength of children's initial biases. The models help to clarify how prefrontal development may lead to advances in flexible thinking.     

Effects of early weaning on anxiety and prefrontal cortical and hippocampal myelination in male and female Wistar rats

We investigated developmental changes in myelin formation in the prefrontal cortex and the hippocampus, and behavioral effects of early weaning in Wistar rats. Early-weaned rats showed decreased numbers of open-arm entries in an elevated plus-maze in both sexes at 4 weeks old; this effect persisted in males, but ceased in females after this age. Expression of myelin basic protein (MBP) showed both age-dependent increases and sex differences; 4-week-old males exhibited higher MBP levels in the hippocampus, whereas 7-week-old males showed lower MBP levels in the prefrontal cortex compared to females of the same age. There was a tendency for group differences from weaning for the 21.5-kDa isoform in the prefrontal cortex. Although these results suggest that male rats are more vulnerable than females to early-weaning effects on anxiety-related behaviors, further detailed analysis is needed to clarify the functional relationship between myelination and anxiety-related behaviors.     

Genes,cognition and brain through a COMT lens

Various genes are known to modulate the delicate balance of dopamine in prefrontal cortex and influence cortical information processing. Catechol-O-methyltransferase (COMT) on chromosome 22q11 is the most widely studied of these genes. Val158Met, a common, functional variant in the coding sequence that increases or decreases the enzymatic activity of the gene has been shown to impact the efficiency of prefrontally-mediated cognition, specifically executive functioning, working memory, fluid intelligence and attentional control. We review the rapidly evolving literature exploring the association between COMT genotype and cognitive performance, and illustrate how this polymorphism has served a pivotal role in characterizing various interacting dimensions of complexity in the relationship between genes and cognition. We review how Val158Met has been used to help develop and validate behavioral and neurophysiological phenotypes, as a critical tool in dissecting overlapping neural functional systems and exploring interactions within and between genes, and in exploring how gene effects on cognition are modulated by environmental, demographic and developmental factors. Despite the impressive range of findings, the COMT story is also a bracing reminder of how much work remains to translate this knowledge into practical clinical applications.     

The ontogeny of exploratory behavior in male and female adolescent rats (Rattus norvegicus)

During adolescence, rats gain independence from their mothers and disperse from the natal burrow, with males typically dispersing further than females. We predicted that, if dispersal patterns are associated with responsiveness to novelty, exploratory behavior in novel environments would increase across adolescence, and males would explore more than females. Alternatively, females might explore more than males, if females are more motivated than males to learn about the immediate environment or if females have poorer spatial abilities than males. Twenty‐five male and 21 female rats were exposed to two novel environments (open field and elevated plus‐maze) during early, mid‐, or late adolescence. Total locomotion and amount of exploration directed towards aversive areas increased across adolescence, even when body weight was included as a covariate. Female adolescents locomoted more and spent more time exploring aversive areas than males. Developmental changes in neural function potentially underlie age and sex differences in exploratory behavior. © 2009 Wiley Periodicals, Inc. Dev Psychobiol 51: 513–520, 2009     

Repeated MDMA ("Ecstasy") exposure in adolescent male rats alters temperature regulation, spontaneous motor activity, attention, and serotonin transporter binding

Previous research in our laboratory found that repeated exposure of adolescent rats to 3,4-methylenedioxymethamphetamine (MDMA) impaired working memory and reduced anxiety. The present experiment extended these findings by investigating the physiological, behavioral, and neurotoxic effects of a modified MDMA treatment regimen. Male Sprague-Dawley rats received 5 mg/kg of MDMA hourly for a period of 4 hr on every fifth day from postnatal day 35-60. Acute effects of the MDMA treatment included hypothermia, serotonin syndrome behavior, and ejaculation. Body weight gain was attenuated by repeated drug administration. The animals completed anxiety and working memory tests beginning 4 days after the final MDMA dose. MDMA altered habituation to the open-field, increased locomotor activity in the elevated plus-maze, decreased attention in the novel object-recognition test, and reduced serotonin transporter binding in the neocortex. These results indicate that repeated exposure to a relatively moderate MDMA dose during adolescence produces later changes in behavior and neurochemistry.     

Enhanced stress responses in adolescent versus adult rats exposed to cues of predation threat, and peer interaction as a predictor of adult defensiveness

Development of the hypothalamic-pituitary-adrenal (HPA) axis is influenced by external factors during early life in mammals, which optimizes adult function for predicted conditions. We have hypothesized that adolescence represents a sensitive period for the development of some aspects of adult stress response regulation. This was based on prior work showing that repeated exposure of rats to a stressor across an adolescent period increases fearfulness in a novel environment in adulthood and results in lower levels of dopamine receptor subtype-2 protein in prefrontal cortex. Here, we further our investigation of both acute and long-term effects of repeated adolescent stressor exposure on physiological (i.e., corticosterone) and behavioral (i.e., defensive behavior) measures of stress responding in male and female rats. Furthermore, we compared outcomes with those following identical manipulations administered in early adulthood and found that animals exposed to cues of predation threat during adolescence showed the most robust defensive responses to a homotypic stressor encountered in adulthood. Peer interaction during control manipulation in adolescence was identified as an important individual characteristic mediating development of adult defensive strategies.     

Rewarding properties of social interactions in adolescent and adult male and female rats: impact of social versus isolate housing of subjects and partners

Social interactions have been shown to be rewarding for adolescent and adult rats; however, there has been little emphasis on comparing the strength of the rewarding value of social stimuli across ontogeny. Since age differences in social interactions may vary with sex or housing circumstances, the present study assessed social conditioned place preference (CPP) in adolescent and adult male and female Sprague-Dawley rats housed either socially or in isolation and conditioned with either group-housed or isolate-housed partners. Isolated animals of both sexes and ages demonstrated social CPP, with the strongest preference emerging in adolescent males. Social CPP was not evident in group-housed adults whereas group-housed adolescents developed a preference for the compartment previously paired with similarly housed partners; however, when socially housed adolescents were conditioned with isolated partners, social CPP did not emerge. Age differences in social CPP may reflect age-related neural alterations in brain systems implicated in regulation of social behavior.     

Effects of physical exercise on ADHD-like behavior in male and female adolescent spontaneously hypertensive rats

The present study examined the effects of exercising (voluntary wheel running) during adolescence on attentional function in male and female spontaneously hypertensive rats (SHRs), a commonly used animal model of attention-deficit/hyperactivity disorder (ADHD). Once rats reached adulthood, they received one session in which a light was presented 12 times but not reinforced, followed by training sessions in which the light was paired with a food reward. Male and female SHRs that had access to running wheels exhibited levels of unconditioned orienting behavior that were similar to Wistar-Kyoto rats (normo-active control) while SHRs that did not have access to running wheels exhibited higher levels of unconditioned orienting behavior. When the light was later paired with food there were no differences between the groups of male rats, but exercising female SHRs exhibited a decrease in conditioned food cup behavior. Consistent with their established phenotype, SHR rats exhibited more locomotor activity during an open field exploration session than WKY rats, but there was no relationship between orienting behavior and locomotor activity. Together these data suggest that physical exercise during adolescence can benefit attentional capabilities.     

Delayed alternation performance in rats following recovery from early iron deficiency

Early iron deficiency (ID) is one of the most common nutrient deficiencies in both developed and developing countries. This condition has been linked to perturbations in myelin formation, alterations of monoamine neurotransmitter systems particularly in the striatum, and deficits in energy metabolism particularly in the hippocampus (HP) and prefrontal cortex (PFC) in rats. Early ID has also been traced to long-term behavioral consequences in children in domains linked to these neuropathologies. The current experiment assesses formerly iron deficient (FID) adult rats on a delayed alternation (DA) task — a procedure thought to be sensitive to PFC dysfunction. Rat dams were started on an iron deficient chow at gestational day (G) 2 and maintained on this diet until postnatal day (P) 7; behavioral training began at P 65 when animals were iron replete. FID animals exhibited accelerated acquisition (p = 0.002) and fewer errors (p = 0.003) on the DA task compared to controls. These findings may reflect an imbalance between hippocampal and prefrontal modulation of this behavior most likely emanating from long-term hippocampal disinhibition by early ID that persists in spite of early iron treatment from P 7.     

Prenatal cocaine exposure increases anxiety,impairs cognitive function and increases dendritic spine density in adult rats: influence of sex

Cocaine exposure during pregnancy can impact brain development and have long-term behavioral consequences. The present study examined the lasting consequences of prenatal cocaine (PN-COC) exposure on the performance of cognitive tasks and dendritic spine density in adult male and female rats. From gestational day 8 to 20, dams were treated daily with 30 mg/kg (ip) of cocaine HCl or saline. At 62 days of age, offspring were tested consecutively for anxiety, locomotion, visual memory and spatial memory. PN-COC exposure significantly increased anxiety in both sexes. Object recognition (OR) and placement (OP) tasks were used to assess cognitive function. Behavioral tests consisted of an exploration trial (T1) and a recognition trial (T2) that were separated by an inter-trial delay of varying lengths. Male PN-COC subjects displayed significantly less time investigating new objects or object locations during T2 in both OR and OP tasks. By contrast, female PN-COC subjects exhibited impairments only in OR and only at the longest inter-trial delay interval. In addition, gestational cocaine increased dendritic spine density in the prefrontal cortex and nucleus accumbens in both genders, but only females had increased spine density in the CA1 region of the hippocampus. These data reveal that in-utero exposure to cocaine results in enduring alterations in anxiety, cognitive function and spine density in adulthood. Moreover, cognitive deficits were more profound in males than in females.     

Neural substrates of manipulation in visuospatial working memory

The present study aimed to investigate in humans whether similar neuronal mechanisms underlie the manipulation and active processing of visual and visuospatial stimuli. Simultaneous and successive mental rotation and identity judgment of 2-D matrices and 3-D cube figures were contrasted using functional magnetic resonance imaging (fMRI). Results demonstrate that activation patterns during mental rotation with low working memory demands differ depending on stimulus type (2-D vs. 3-D). Comparison of simultaneous mental rotation of matrices and 3-D cubes resulted in activation of frontal as well as inferior and superior parietal cortices. The opposite contrast (mental rotation of 3-D cubes vs. 2-D matrices) yielded only frontal cortex activation. The findings also yield evidence for converging, overlapping activation patterns for 2-D and 3-D stimuli if working memory demands are increased. Results are discussed within the framework of current working memory models.     

Activity of primate orbitofrontal and dorsolateral prefrontal neurons: task-related activity during an oculomotor delayed-response task

The orbitofrontal cortex (OFC) has strong reciprocal connections to the dorsolateral prefrontal cortex (DLPFC), which is known to participate in spatial working memory processes. However, it is not known whether or not the OFC also participates in spatial working memory and whether the OFC and DLPFC contribute equally to this process. To address these issues, we collected single-neuron activity from both areas while a monkey performed an oculomotor delayed-response task, and compared the characteristics of task-related activities between the OFC and DLPFC. All of the task-related activities observed in the DLPFC were also observed in the OFC. However, the proportion and response characteristics of task-related activities were different between the two areas. While most delay-period activity observed in the DLPFC was directionally selective and showed tonic sustained activation, most delay-period activity observed in the OFC was omni-directional and showed gradually increasing activity. Reward-period activity was predominant among task-related activities in the OFC. The proportion of neurons showing reward-period activity was significantly higher in the OFC than in the DLPFC. These results suggest that, although both the OFC and DLPFC participate in spatial working memory processes, the OFC is related more to the expectation and the detection of reward delivery, while the DLPFC is related more to the temporary maintenance of spatial information and its processing.     

Adolescent female rats are more resistant than males to the effects of early stress on prefrontal cortex and impulsive behavior

We tested the hypothesis that adolescent Sprague–Dawley females may be more resistant than males to display impulsive behavior and lower prefrontal cortex thickness after mother‐infant separation (MS). Starting at postnatal day 2 (P2), the MS group was separated 6 hr/day and the early handled (EH) group 15 min/day for 10 days, and another group was standard facility reared (SFR). Subjects were examined for novel open‐field activity (P28), light–dark apparatus (P29), familiar open‐field (P30) and frontal cortical thickness. This protocol resulted in impulsive behavior in MS rats relative to EH and SFR, but this effect was less pronounced in females than males. MS affected the two sexes differently in terms of decreased prefrontal cortex dorsoventral thickness, with this effect being significant in males but not females. Neuroanatomical and behavioral documentation that adolescent females are more resistant than males to ADHD‐like effects of maternal separation have not been previously reported. © 2009 Wiley Periodicals, Inc. Dev Psychobiol 51: 277–288, 2009