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• GABAergic and glutamatergic terminals express SV2A in a layer-dependent manner. • GABAergic and glutamatergic terminals express SV2A in an age-dependent manner. • The SV2A protein may improve the excitatory/inhibitory balance in the hippocampus. The function of synaptic vesicle protein 2A (SV2A) has not been clearly identified, although it has an essential role in normal neurotransmission. Changes in SV2A expression have been linked to several diseases that could implicate an imbalance between excitation and inhibition, such as epilepsy. Although it is known that SV2A expression is necessary for survival, SV2A expression and its relationship with γ-aminobutyric acid (GABA) and glutamate neurotransmitter systems along development has not been addressed. This report follows SV2A expression levels in the rat hippocampus and their association with glutamatergic and GABAergic terminals along postnatal development. Total SV2A expression was assessed by real time PCR and western blot, while immunofluorescence was used to identify SV2A protein in the different hippocampal layers and its co-localization with GABA or glutamate vesicular transporters. SV2A was dynamically regulated along development and its association with GABA or glutamate transporters varied in the different hippocampal layers. In the principal cells layers (granular and pyramidal), SV2A protein was preferentially localized to GABAergic terminals, while in the hilus and stratum lucidum SV2A was associated mainly to glutamatergic terminals. Although SV2A was ubiquitously expressed in the entire hippocampus, it established a differential association with excitatory or inhibitory terminals, which could contribute to the maturation of excitatory/inhibitory balance. [ABSTRACT FROM AUTHOR]

• Selectively-bred high- and low-responder rats differed in affect in early life. • High- and low-responder rats showed different opioid gene expression. • Neonatal administration of FGF2 decreased maternal separation distress calls at P11. • Neonatal FGF2 increased positive affect during exploration and play in juveniles. • Neonatal FGF2 altered expression of the mu opioid receptor and prodynorphin. In previous studies, early-life fibroblast growth factor-2 (FGF2) administration conferred resilience to developing anxiety-like behavior in vulnerable animals in adulthood. To follow up on this work, we administered FGF2 the day after birth to animals that differ in emotional behavior and further explored its long-term effects on affective behavior and circuitry. Selectively-bred "high responder" rats (bHRs) exhibit low levels of anxiety-like and depression-like behavior, whereas selectively-bred "low responders" (bLRs) display high levels of anxiety-like and depression-like behavior. We found that early-life administration of FGF2 decreased negative affect in bLRs during the early post-natal period, as indexed by 40 kHz ultrasonic vocalizations (USVs) in response to a brief maternal separation on PND11. FGF2 also increased positive affect during the juvenile period, as measured by 50 kHz USVs in response to heterospecific hand play ("tickling") after weaning. In general, we found that bHRs produced more 50 kHz USVs than bLRs. In adulthood, we measured opioid ligand and receptor expression in brain regions implicated in USV production and affect regulation by mRNA in situ hybridization. Within multiple affective brain regions, bHRs had greater expression of the mu opioid receptor than bLRs. FGF2 increased mu opioid expression in bLRs. The bLRs had more kappa and less delta receptor expression than bHRs, and FGF2 increased prodynorphin in bLRs. Our results provide support for further investigations into the role of growth factors and endogenous opioids in the treatment of disorders characterized by altered affect, such as anxiety and depression. [ABSTRACT FROM AUTHOR]

Highlights • Neonatal predator odor exposure alters opioid receptor mRNA levels. • Changes in expression are observed in the neonatal and juvenile nucleus accumbens. • These changes in expression are sex-specific, restricted to females. • Changes in expression do not occur in the neonatal nor juvenile amygdala. • These changes could lead to sex-specific responses to opioids. Abstract Early life stress (ELS) strongly impacts mental health, but little is known about its interaction with biological sex and postnatal development to influence risk and resilience to psychopathologies. A number of psychiatric disorders, such as social anhedonia and drug addiction, involve dysfunctional opioid signaling; moreover, there is evidence for differential central opioid function in males vs. females. The present study examined opioid receptor gene expression in the nucleus accumbens (NAc) and amygdala of male and female rats subjected to a neonatal predator odor exposure (POE) paradigm to model ELS. Brain tissue was collected at two developmental time points: neonatal and juvenile. Results showed that, following the neonatal POE experience, opioid receptor mRNA levels in the NAc were differentially regulated at the neonatal and juvenile time points. POE downregulated neonatal mu- and kappa-opioid receptor mRNA levels in neonatal females, but upregulated mu- and delta-opioid receptor mRNA levels in juvenile females. Intriguingly, POE had no significant effect on NAc opioid receptor mRNA levels in males at either time point, indicating that the impact of POE on opioid system development is sex-dependent. Finally, POE failed to alter amygdalar opioid receptor gene expression in either sex at either time-point. The spatiotemporally- and sex-specific impact of ELS within the developing brain may confer differential risk or resilience for males and females to develop atypical opioid-regulated behaviors associated with conditions such as depression and addiction. [ABSTRACT FROM AUTHOR]

Seizures induced by fever are the most prevalent age-specific seizures in infants and young children. Whether they result in long-term sequelae such as neuronal loss and temporal lobe epilepsy is controversial. Prospective studies of human febrile seizures have found no adverse effects on the developing brain. However, adults with temporal lobe epilepsy and associated limbic cell loss frequently have a history of prolonged febrile seizures in early life. These critical issues may be resolved using appropriate animal models. Published models of hyperthermic seizures have used 'adolescent' and older rats, have yielded a low percentage of animals with actual seizures, or have suffered from a high mortality, rendering them unsuitable for long-term studies. This article describes the establishment of a model of febrile seizures using the infant rat. Hyperthermia was induced by a regulated stream of mildly heated air, and the seizures were determined by both behavioral and electroencephalographic (EEG) criteria. Stereotyped seizures were generated in 93.6% of 10-11-day-old rats. EEG correlates of these seizures were not evident in cortical recordings, but were clearly present in depth recordings from the amygdala and hippocampus. Prolonged febrile seizures could be induced without burns, yielding a low mortality (11%) and long-term survival. In summary, in infant rat paradigm of EEG-confirmed, hyperthermia-induced seizures which is suitable for long-term studies is described. This model should be highly valuable for studying the mechanisms and sequelae of febrile seizures.

The ontogeny of corticotropin-releasing hormone (CRH) receptor messenger ribonucleic acid (mRNA) in rat brain, using in situ hybridization, is the focus of this study. The developmental profile of CRH receptor using binding assays and receptor autoradiography has been reported, but may be confounded by the presence of a binding protein. The recent cloning of the rat CRH receptor gene has permitted the use of in situ hybridization histochemistry to map the distribution of cells expressing CRH receptor mRNA in the developing brain. We used antisense 35S-labeled oligodeoxynucleotide probes for the two reported splice-variants of the CRH receptor mRNA, which yielded essentially identical localization patterns. CRH receptor mRNA was clearly detectable in infant brain starting on the second postnatal day. Signal in hippocampal CA1, CA2 and CA3a increased to 300-600% of adult levels by postnatal day 6 with a subsequent gradual decline. In the amygdala, in contrast, CRH receptor mRNA abundance increased steadily between the second and the ninth postnatal days, to levels twice higher than those in the adult. In the cortex, CRH receptor mRNA levels were high on postnatal day 2 and decreased to adult levels by day 12. Transient signal over the hypothalamic paraventricular nucleus, observed on the second postnatal day, was not evident at older ages. These results demonstrate robust synthesis of CRH receptor as early as on the second postnatal day and unique region-specific developmental profiles for CRH receptor gene expression.

Mechanisms controlling the synthesis of corticotropin releasing hormone (CRH) in neonatal rats, and the ontogeny of glucocorticoid (GC) feedback control of hypothalamic CRH remain unknown. Specific issues are whether stress induces up-regulation of CRH gene expression during the first postnatal week, and the role of GC feedback, at the hypothalamic level, in the stress-hyporesponsive period. We studied the ontogeny of the negative feedback regulation of CRH gene expression by GC in the paraventricular nucleus (PVN). We implanted chronic cannulae containing a GC-receptor antagonist, RU 38486, in rats on postnatal days 3 to 13. Three days later, animals were sacrificed, and brains were analyzed for CRH-messenger RNA (CRH-mRNA), using semi-quantitative in situ hybridization. Animals implanted with cholesterol-containing cannulae served to evaluate the stressful effect of implantation on CRH-mRNA abundance. The presence of GC receptor messenger RNA (GR-mRNA) in the PVN of neonatal rats was also determined. RU 38486 did not increase CRH-mRNA abundance during the first postnatal week, despite the presence of GR-mRNA in the PVN. Chronic-implantation stress also failed to increase CRH synthesis. CRH gene expression in the PVN was enhanced in infant rats implanted with RU-38486 on postnatal day 9 or later. Cholesterol implantation on days 9, 10 (but not later), resulted in increased PVN-CRH-mRNA. Thus, CRH-mRNA is up-regulated by chronic blockade of GC receptors only subsequent to the eighth postnatal day. Furthermore, such blockade does not affect the response of CRH-mRNA to chronic stress in the neonatal rat.

Little is known about what exactly differentiates metacognitive processes from ordinary cognitive processes particularly early in development, and the underlying developmental aspects. To examine the time-course of metacognition, the present study investigated the neural underpinnings of judgments of learning (JoLs) and compared them with control judgments, using an event-related potentials (ERP) design. During ERP recording, children age seven to eight were presented with cue-target picture pairs and instructed to learn these pairs. After each pair, they either had to make a JoL (assess the likelihood of remembering the target when only presented with the cue) or a colour judgment (indicate whether the colour yellow had been present in one of the two pictures presented earlier). Results revealed a late slow wave divergence maximal pronounced from 550 ms to 950 ms post-stimulus that distinguished between JOLs and colour judgments. Over centro-parietal areas, JoLs showed a more negative going slow wave compared to the colour judgments, and this pattern was independent of performance. The results are in support of theories that assume a distinction between metacognitive and cognitive processes. [ABSTRACT FROM AUTHOR]

Repeated exposure to psychostimulants during adolescence produces long-lasting changes in behavior that may be mediated by disrupted development of the mesocorticolimbic dopamine system. Here, we tested this hypothesis by assessing the effects of amphetamine (AMPH) and dopamine receptor-selective drugs on behavior and neuron activity in the prelimbic region of the medial prefrontal cortex (PFC). Adolescent male, Sprague-Dawley rats were given saline or 3 mg/kg AMPH between postnatal day (P) 27 and P45. In Experiment 1, locomotor behavior was assessed during adulthood following challenges with a dopamine D 1 (SKF 82958) or D 2 (quinpirole) receptor-selective agonist. In Experiment 2, pre-exposed rats were challenged during adulthood with AMPH and a D 1 (SKF 83566) or D 2 (eticlopride) receptor-selective antagonist. In Experiment 3, the activity of putative pyramidal cells in the prelimbic cortex was recorded as rats behaved in an open-field arena before and after challenge injections with AMPH and one of the antagonists. We found that compared to controls, adolescent pre-exposed rats were more sensitive to the stimulant effects of AMPH and the dopamine receptor agonists, as well as to the ability of the antagonists to reverse AMPH-induced stereotypy. Prelimbic neurons from AMPH pre-exposed rats were also more likely to respond to an AMPH challenge in adulthood, primarily by reducing their activity, and the antagonists reversed these effects. Our results suggest that exposure to AMPH during adolescence leads to enduring adaptations in the mesocorticolimbic dopamine system that likely mediate heightened response to the drug during adulthood. [ABSTRACT FROM AUTHOR]

Maternal smoking has negative long-term consequences on affective behaviors, and in rodents, chronic neonatal nicotine exposure (CNN) results in increased anxiety. In rat pups, acute nicotine stimulation activates brain regions associated with stress and anxiety, but chronic nicotine exposure could desensitize of nicotinic acetylcholine receptors, the molecular target of nicotine. Here, we determined whether CNN affected neuronal activation by an acute nicotine challenge. Using in situ hybridization, we analyzed mRNA expression of the immediate-early genes (IEGs) c-Fos, Arc, Egr-1 and Npas4, which are markers for neuronal activation and implicated in synaptic plasticity. Following CNN (6 mg/kg/day) or control treatment from postnatal day (P)1 to P7, an acute i.p. nicotine (0.7 mg/kg) or saline injection (control) was administered on P8, and brains collected after 30 min. In drug-naive pups, acute nicotine stimulated IEGs expression specifically in brain areas associated with innate anxiety including the paraventricular hypothalamic nucleus, central nucleus of the amygdala (CeA), and locus coeruleus (LC). Following CNN, acute nicotine stimulated IEG expression in all three areas, but activation was significantly reduced in the LC (c-Fos, Egr-1, Npas4), and CeA (c-Fos). Notably, nicotine-induced Npas4 expression was greatly diminished in the LC, which may affect inhibitory synapse formation in noradrenergic neurons. Thus, after CNN, neurons located in areas associated with anxiety brain circuitry maintained responsiveness to nicotine, but tolerance differentially developed to nicotine. In the developing brain, repeated activation by nicotine of areas related to limbic pathways could alter circuit connectivity and increase responsiveness to stress and anxiety later in life. [ABSTRACT FROM AUTHOR]

Different physical or chemical agents, such as noise or alcohol, can induce diverse behavioral and biochemical alterations. Considering the high probability of young people to undergo consecutive or simultaneous exposures, the aim of the present work was to investigate in an animal model if noise exposure at early adolescence could induce hippocampal-related behavioral changes that might be modified after alcohol intake. Male Wistar rats (28-days-old) were exposed to noise (95–97 dB, 2 h). Afterwards, animals were allowed to voluntarily drink alcohol (10% ethanol in tap water) for three consecutive days, using the two-bottle free choice paradigm. After that, hippocampal-related memory and anxiety-like behavior tests were performed. Results show that whereas noise-exposed rats presented deficits in habituation memory, those who drank alcohol exhibited impairments in associative memory and anxiety-like behaviors. In contrast, exposure to noise followed by alcohol intake showed increases in exploratory and locomotor activities as well as in anxiety-like behaviors, unlike what was observed using each agent separately. Finally, lower levels of alcohol intake were measured in these animals when compared with those that drank alcohol and were not exposed to noise. Present findings demonstrate that exposure to physical and chemical challenges during early adolescence might induce behavioral alterations that could differ depending on the schedule used, suggesting a high vulnerability of rat developing brain to these socially relevant agents. [ABSTRACT FROM AUTHOR]

Olfactory glomeruli are the first synaptic site of the olfactory system and are formed by the convergence of axons of the same type of sensory neurons onto the olfactory bulbs of the brain. Although the anatomical organization of glomeruli is conserved across species, their particular role in olfactory processing remains uncertain. We studied the composition and maintenance of glomeruli by means of a genetic model, mI7-IRES- tau GFP knock-in young mice, where the cytoskeleton of sensory neurons expressing the mI7 olfactory receptor is tagged with green fluorescent protein. Animals were continuously exposed to heptaldehyde, a cognate ligand of the mI7 receptor, from postnatal days 5–10. We hypothesized that continuous odorant exposure will induce changes in glomerular morphology, and that this can be recovered if the normal odorant environment is reestablished within the early postnatal period. We assessed changes in the distribution of mI7 axons in glomerular morphology, as well as possible changes in the number of the mI7 olfactory sensory neurons. Following odorant exposure the well-defined convergence of mI7 fibers into a single glomerulus was disrupted, producing numerous neighboring glomeruli partially innervated by mI7 fibers. After the normal odor environment was reestablished the number of glomeruli partially innervated by mI7 fibers decreased significantly. Moreover, we found that multiple supernumerary mI7 glomeruli were formed. Our results confirm the significant role of sensory input in glomerular formation and maintenance. Additionally, we show that the developing olfactory system actively maintains glomerular morphology, suggesting the importance of this for olfactory processing. [ABSTRACT FROM AUTHOR]

We reported previously that monoamine oxidase (MAO) A knockout (KO) mice show increased serotonin (5-hydroxytryptamine, 5-HT) levels and autistic-like behaviors characterized by repetitive behaviors, and anti-social behaviors. We showed that administration of the serotonin synthesis inhibitor para-chlorophenylalanine (pCPA) from post-natal day 1 (P1) through 7 (P7) in MAO A KO mice reduced the serotonin level to normal and reverses the repetitive behavior. These results suggested that the altered gene expression at P1 and P7 may be important for the autistic-like behaviors seen in MAO A KO mice and was studied here. In this study, Affymetrix mRNA array data for P1 and P7 MAO A KO mice were analyzed using Partek Genomics Suite and Ingenuity Pathways Analysis to identify genes differentially expressed versus wild-type and assess their functions and relationships. The number of significant differentially expressed genes (DEGs) varied with age: P1 (664) and P7 (3307) [false discovery rate (FDR) <0.05, fold-change (FC) >1.5 for autism-linked genes and >2.0 for functionally categorized genes]. Eight autism-linked genes were differentially expressed in P1 (upregulated: NLGN3, SLC6A2 ; down-regulated: HTR2C, MET, ADSL, MECP2, ALDH5A1, GRIN3B) while four autism-linked genes were differentially expressed at P7 (upregulated: HTR2B; downregulated: GRIN2D, GRIN2B, CHRNA4 ). Many other genes involved in neurodevelopment, apoptosis, neurotransmission, and cognitive function were differentially expressed at P7 in MAO A KO mice. This result suggests that modulation of these genes by the increased serotonin may lead to neurodevelopmental alteration in MAO A KO mice and results in autistic-like behaviors. [ABSTRACT FROM AUTHOR]

Neural circuit formation is a critical process in brain development. Axon guidance molecules, their receptors, and intracellular mediators are important to establish neural circuits. Collapsin response mediator proteins (CRMPs) are known intercellular mediators of a number of repulsive guidance molecules. Studies of mutant mice suggest roles of CRMPs in dendrite development. However, molecular mechanisms of CRMP-mediated dendritic development remain to elucidate. In this study, we show abnormal orientation of basal dendrites (extension to deeper side) of layer V pyramidal neurons in the cerebral cortex of CRMP4−/− mice. Moreover, we observed severe abnormality in orientation of the basal dendrites of these neurons in double knockout of CRMP1 and 4, suggesting redundant functions of these two genes. Redundant gene functions were also observed in proximal bifurcation phenotype in apical dendrites of hippocampal CA1 pyramidal neurons. These results indicate that CRMP1 and CRMP4 regulate proper orientation of the basal dendrites of layer V neurons in the cerebral cortex. [ABSTRACT FROM AUTHOR]

Ongoing development of the dopamine system during adolescence may provide a partial mechanism for behavioral and psychiatric vulnerabilities. Despite early evidence for a hyperactive adolescent dopaminergic system, recent data suggest that adolescent dopamine may be functionally hypoactive compared to in adults. While this distinction has been established in response to dopaminergic drugs and natural rewards, little is known about age-related differences in cognitive efficacy of dopaminergic drugs. Using a recently established Cued Response Inhibition Task, we tested the effects of acute systemic methylphenidate, commonly known as Ritalin , on response inhibition and response initiation in adolescent and adults rats. First, we replicated previous data that adolescents are able to inhibit a response to a cue on par with adults, but are slower to produce a rewarded response after a stop cue. Next, we observed that methylphenidate modulated response inhibition in adult rats, with low dose (0.3 mg/kg) improving inhibition, and high dose (3 mg/kg) impairing performance. This dose-response pattern is commonly observed with psychostimulant cognitive modulation. In adolescents, however, methylphenidate had no effect on response inhibition at any dose. Latency of response initiation after the stop cue was not affected by methylphenidate in either adult or adolescent rats. These data establish that dose-response of a commonly prescribed psychostimulant medication is different in adolescents and adults. They further demonstrate that healthy adolescent response inhibition is not as sensitive to psychostimulants as in adults, supporting the idea that the dopamine system is hypoactive in adolescence. This article is part of a Special Issue entitled SI: Adolescent plasticity . [ABSTRACT FROM AUTHOR]

[Display omitted] • In utero exposure to VPA results in an increased risk of ASD in humans. • VPA exposure resulted in fewer neurons in vestibular nuclei and fewer CB puncta. • VPA exposure resulted in impaired performance on motor tasks. • VPA exposure resulted in delayed VEMPs and horizontal nystagmus. Autism spectrum disorder (ASD) is a neurodevelopmental disorder that includes several key neuropathological changes and behavioral impairments. In utero exposure to the anti-epileptic valproic acid (VPA) increases risk of an ASD diagnosis in human subjects and timed in utero exposure to VPA is a clinically relevant animal model of ASD. Many human subjects with ASD have cerebellar hypoplasia, fewer Purkinje cells, difficulties with balance, ophthalmic dysfunction and abnormal responses to vestibular stimulation and such vestibular difficulties are likely under reported in ASD. We have recently shown that animals exposed to VPA in utero have fewer neurons in their auditory brainstem, reduced axonal projections to the auditory midbrain and thalamus, reduced expression of the calcium binding protein calbindin (CB) in the brainstem and cerebellum, smaller and occasionally ectopic cerebellar Purkinje cells and ataxia on several motor tasks. Based on these findings, we hypothesized that in utero VPA exposure similarly impacts structure and function of the vestibular brainstem. We investigated this hypothesis using quantitative morphometric analyses, immunohistochemistry for CB, a battery of vestibular challenges, recording of vestibular-evoked myogenic potentials and spontaneous eye movements. Our results indicate that VPA exposure results in fewer neurons in the vestibular nuclei, fewer CB-positive puncta, difficulty on certain motor tasks, longer latency VEMPs and significantly more horizontal eye movements. These findings indicate that the vestibular nuclei are impacted by in utero VPA exposure and provide a basis for further study of vestibular circuits in human cases of ASD. [ABSTRACT FROM AUTHOR]

RNA-binding protein activities are highly regulated through protein levels, intracellular localization, and post-translation modifications. During development, mRNA processing of specific gene sets is regulated through manipulation of functional RNA-binding protein activities. The impact of altered RNA-binding protein activities also affects human diseases in which there are either a gain-of-function or loss-of-function causes pathogenesis. We will discuss RNA-binding proteins and their normal developmental RNA metabolism and contrast how their function is disrupted in disease. This article is part of a Special Issue entitled SI:RNA Metabolism in Disease . [ABSTRACT FROM AUTHOR]

Neurensin1 (Nrsn1) gene, highly specific to neurons, has been considered to play a role in neurite growth during neuronal development and regeneration in mice. Intense expression of Nrsn1 was found particularly in projecting neurons like retinal ganglion cells and spinal motor neurons, suggesting that Neurensin1 is needed for active neurite growth. In the present study we cloned chick Nrsn1 gene and produced an antibody against cNrsn1 to examine Nrsn1 localization in the chick brain, since the chick is a suitable animal model for the study of developmental neurobiology. We found that there are neurons intensely stained for Nrsn1 antibody localized in the optic tectum, the cerebellum and the brain stem. These neurons are large in size and considered to be projecting neurons. In the cerebellum, Purkinje cells are the only one type of neurons stained for Nrsn1. During Purkinje cell development the arborized dendrites and axons become intensely stained at stages E17-18. A siRNA gene knock down was applied to the cultured embryonic cerebellar tissues and the result showed that Nrsn1 has an important role in dendrite formation of Purkinje cells. These findings suggest that Neurensin1 is also involved in neural development in the chick brain and that the embryonic chick brain is a good model to disclose the molecular and physiological functions of Nrsn1. [ABSTRACT FROM AUTHOR]

Objective To investigate whether high performance on college preparedness tests at 18 years of age can be predicted from brain activation patterns during narrative comprehension at 5–7 years of age. Methods In this longitudinal study, functional MRI data during an auditory narrative-comprehension task were acquired from 15 children (5–7 years of age) who also provided their American College Testing (ACT) scores at the age of 18 years. Active voxels during the narrative-comprehension task were correlated with both composite ACT scores and the reading-comprehension component of the exam. Results Higher composite ACT scores and behavioral scores for reading comprehension were positively correlated with greater activation in frontal and anterior brain regions during the narrative-comprehension task. Conclusions Our results suggest that neural circuits supporting higher ACT performance are predictable from a narrative-comprehension task at the age of 5–7 years. This supports a critical role for the anterior cingulate cortex, which is a part of the cingulo-opercular cognitive-control network early in development, as a facilitator for better ACT scores. This study highlights that shared neural circuits that support overall ACT performance and neural circuits that support reading comprehension both rely on neural circuits related to narrative comprehension in childhood, suggesting that interventions involving narrative comprehension should be considered for individuals with reading and other academic difficulties. [ABSTRACT FROM AUTHOR]

As the receptor of myelin associated inhibitory factors Nogo receptor 1 (NgR1) plays an important role in central nervous system (CNS) injury and regeneration. It is found that NgR1 complex acts in neurons to transduce the signals intracelluarly including induction of growth cone collapse, inhibition of axonal regeneration and regulation of nerve inflammation. In recent studies, NgR1 has also been found to be expressed in the microglia. However, NgR1 expressed in microglia in the developing nervous systems and following CNS injury have not been widely investigated. In this study, we detected the expression and cellular localization of NgR1 in microglia during development and following traumatic brain injury (TBI) in mice. The results showed that NgR1 was mainly expressed in microglia during embryonic and postnatal periods. The expression levels peaked at P4 and decreased thereafter into adulthood, while increased significantly with aging representatively at 17 mo. On the other hand, there was no significant difference in the number of double positive NgR1 + Iba1 + cells between normal and TBI group. In summary, we first detected the expression of NgR1 in microglia during development and found that NgR1 protein expression increased significantly in microglia with aging. These findings will contribute to make a foundation for subsequent study about the role of NgR1 expressed in microglia on the CNS disorders. [ABSTRACT FROM AUTHOR]

Studies of human brain malformations, such as lissencephaly and double cortex, have revealed the importance of neuronal migration during cortical development. Afadin, a membrane scaffolding protein, regulates the formation of adherens junctions (AJs) and cell migration to form and maintain tissue structures. Here, we report that mice with dorsal telencephalon-specific ablation of afadin gene exhibited defects similar to human double cortex, in which the heterotopic cortex was located underneath the normotopic cortex. The normotopic cortex of the mutant mice was arranged in the pattern similar to the cortex of the control mice, while the heterotopic cortex was disorganized. As seen in human patients, double cortex in the mutant mice was formed by impaired neuronal migration during cortical development. Genetic ablation of afadin in the embryonic cerebral cortex disrupted AJs of radial glial cells, likely resulting in the retraction of the apical endfeet from the ventricular surface and the dispersion of radial glial cells from the ventricular zone to the subventricular and intermediate zones. These results indicate that afadin is required for the maintenance of AJs of radial glial cells and that the disruption of AJs might cause an abnormal radial scaffold for neuronal migration. In contrast, the proliferation or differentiation of radial glial cells was not significantly affected. Taken together, these findings indicate that afadin is required for the maintenance of the radial glial scaffold for neuronal migration and that the genetic ablation of afadin leads to the formation of double cortex. [ABSTRACT FROM AUTHOR]

Extensive research implicates disturbed immune function and development in the etiology and pathology of schizophrenia. In addition to reviewing evidence for immunological factors in schizophrenia, this paper discusses how an emerging model of atypical immune function and development helps explain a wide variety of well-established – but puzzling – findings about schizophrenia. A number of theorists have presented hypotheses that early immune system programming, disrupted by pre- and perinatal adversity, often combines with abnormal brain development to produce schizophrenia. The present paper focuses on the hypothesis that disruption of early immune system development produces a latent immune vulnerability that manifests more fully after puberty, when changes in immune function and the thymus leave individuals more susceptible to infections and immune dysfunctions that contribute to schizophrenia. Complementing neurodevelopmental models, this hypothesis integrates findings on many contributing factors to schizophrenia, including prenatal adversity, genes, climate, migration, infections, and stress, among others. It helps explain, for example, why (a) schizophrenia onset is typically delayed until years after prenatal adversity, (b) individual risk factors alone often do not lead to schizophrenia, and (c) schizophrenia prevalence rates actually tend to be higher in economically advantaged countries. Here we discuss how the hypothesis explains 10 key findings, and suggests new, potentially highly cost-effective, strategies for treatment and prevention of schizophrenia. Moreover, while most human research linking immune factors to schizophrenia has been correlational , these strategies provide ethical ways to experimentally test in humans theories about immune function and schizophrenia. This article is part of a Special Issue entitled SI: Neuroimmunology in Health And Disease . [ABSTRACT FROM AUTHOR]

Retinocollicular connections form precise topographical maps that are normally completed through the selective elimination of misplaced axons and the stabilization of topographically ordered axon terminals during early development. Omega-3 fatty acids, acquired exclusively through the diet, and its main metabolite, docosahexaenoic acid (DHA), are involved in brain development and synaptic maturation. We have previously shown that the nutritional restriction of omega-3/DHA results in abnormal retinocollicular topographical fine-tuning. Therefore, we studied the role of omega-3 fatty acids nutritional supplementation and the developmental time windows during which this postnatal supplementation would restore normal topographical maps in the visual system. Female rats and their litters were chronically fed with either control (soy oil) or restricted omega-3 (coconut oil) diets. Fish oil supplementation was introduced between either postnatal day (PND) 7-13, PND7-28 or PND21-42. At PND13, PND28 or PND42, animals received an anterograde eye injection of a neuronal tracer to visualize retinocollicular axons. Confirming previous observations we found that an omega-3/DHA deficiency resulted in an abnormally high innervation density of retinal axons at the visual layers of the superior colliculus (SC). Although a short-term fish oil supplementation between PND7-13 could not restore normal retinocollicular topography, an extended treatment between PND7-28 completely recovered normal innervation densities of retinotectal axons. However, a late onset supplementation protocol, between PND28-42, was no longer effective in the restoration of the abnormal topographical pattern induced by an early omega-3 nutritional malnutrition. The results suggest a critical period for omega3/DHA dietary intake for the proper development of visual topographical maps. [ABSTRACT FROM AUTHOR]

Thromboxane A2 (TXA2) is a central prostanoid in the cardiovascular system and is a crucial mediator of vascular homeostasis and platelet aggregation. In this study, we report a novel role for TXA2 in neurite outgrowth. TXA2 receptor is expressed in rat cortical neurons, and treatment with the TXA2 agonist U-46619 promotes neurite outgrowth in a concentration-dependent manner. We investigated the molecular mechanism underlying U-46619-induced neurite outgrowth in cortical neurons. Blockade of the phosphorylation of mitogen activated protein kinase (MAPK) prevents U-46619-mediated neurite outgrowth. These data indicates that TXA2 functions as a positive regulator of neurite outgrowth via a mechanism dependent on MAPKs in cortical neurons. [ABSTRACT FROM AUTHOR]

In WAG/Rij rat genetic model of absence epilepsy, the first spike-wave discharges (EEG hallmark of absence epilepsy) are known to appear after puberty, and their incidence increases with age. WAG/Rij rats are known to have a genetic predisposition to absence epilepsy, and further development of epilepsy might be influenced by epigenetic factors. This preliminary study examined the effect of early postnatal factors on the incidence of epileptic spike-wave discharges in adulthood. The newborn WAG/Rij rats were fostered by Wistar dams (from birth throughout the weaning age), and their EEG was examined continuously from 5 to 13 months of age. It was found that the number and duration of absence seizures was reduced in WAG/Rij rats adopted by Wistar dams as compared with the age-matched control WAG/Rij rats nursed by their own mothers. These data indicate that natural (epigenetic) factors, such as maternal care during suckling period, affect development of seizure activity in genetically prone subjects. It is suggested that improvement of primarily care-giving environment in subjects with genetic predisposition to absence epilepsy is a way to reduce epileptic activity in later life. [ABSTRACT FROM AUTHOR]

• Chronic perinatal subconvulsive activity leads to sex-related differences in pre-juvenile behavior. • Naïve weanling rat pups exhibit functional alternation asymmetry bias to the left. • Subconvulsive activity removes alternation asymmetry bias. • Convulsive activity induces a compulsive switch in functional asymmetry to the right. • Immobility reduction occurs after subconvulsive or convulsive activity only in males. • Sex differences disappear following single or multiple tonic-clonic convulsion(s). Chronic subconvulsive activity in early life leads to sex-related autistic-like deficits in handling, object recognition, and social performance in pre-pubertal rats. Since autism and epilepsy are common neurodevelopmental disorders with high coincidence, we tested whether early-life chronic subconvulsive activity compared to convulsive activity alters handling, spatial memory, lateralization, coping strategy and the seizure threshold in a sex-dependent manner. A hypothesis is that convulsive seizures may alter sex differences induced by subconvulsive (SC) activity. Serial subconvulsive doses of kainic acid (KA) were administered postnatally (0.25–1 mg/kg) for 15 days to induce the chronic subconvulsive phenotype (SC group). Age-matched controls and a subset of SC pups were exposed to a convulsive dose of KA (KA and SC + KA groups; 7.5 mg/kg) or flurothyl vapors. In our open handling test, controls and the ASD groups escaped to a similar degree whereas after convulsive seizures, the pups exhibited freezing behavior; no escapes occurred. In the spontaneous alternating T-Maze control males and females entered the left arm with higher frequency. The SC males but not SC females entered left and right arms to a similar degree; alternation rates were reduced to chance revealing a sex difference. However, in KA and SC + KA groups, there was a sharp loss of spontaneous alternation rates. The rapid repetitive entries shifted to the right in both sexes possibly be due to hippocampal injury and changes in network activity induced by status epilepticus. In the forced swim test (FST), control and CS females were more active than corresponding males. After convulsions, immobility was reduced and vertical mobility was increased in SC and SC + KA males suggesting an elevated coping strategy compared to females. Onset and severity of KA induced status epilepticus was delayed in SC males and females possibly due to desensitization of KA receptors. Following flurothyl exposure, control males had faster onset of twitches and clonic seizures than control females which disappeared after the sub-convulsive pre-treatment. Data suggest that behavioral manifestations are more readily detectable between males and females when low levels of hyperexcitation are present chronically in early postnatal development but diminished after tonic-clonic convulsions persist. Therefore, therapeutic interventions may benefit patients if initiated upon the initial onset of sex-related autistic pathologies, particularly in males, which may reduce subsequent vulnerability to seizures. [ABSTRACT FROM AUTHOR]

Nitric oxide synthase (NOS) is highly expressed in the hypothalamus, and nitric oxide (NO) specifically contributes to the regulation of neuronal activity within distinct hypothalamic regions. We studied the long-lasting effects of developmental exposure to low doses of organophosphate chlorpyrifos (CPF) and diazinon (DZN) on the expression of NOS in the hypothalamic subnuclei that subserve neuroendocrine, autonomic and cognitive functions. A daily dose of 1 mg/kg of either CPF or DZN was administered to developing rats during gestational days 15–18 or postnatal days (PND) 1–4. Brain sections from PND 60 rats were processed using NADPH-diaphorase (NADPH-d) and neuronal NOS (nNOS) immunohistochemistry. The number of labeled neurons and the optical density (OD) were assessed in the supraoptic (SON), paraventricular (PVN), medial septum, vertical limb, and horizontal limb of the diagonal band. Developmental exposure to organophosphates increased the number of labeled neurons and OD in different subnuclei in the hypothalamus without gender selectivity. The effect on OD was more pronounced and was significant for more cases. Prenatal exposure to CPF and DZN significantly increased the OD in all regions studied with the exception of PVN. Neonatal exposure to DZN also consistently increased OD in all studied subnuclei. For rats that treated with CPF during early postnatal period, this effect was statistically significant only for the SON and PVN. These findings suggest that overexpression of NOS in the hypothalamus may contribute to the mechanisms inducing or compensating for endocrine, autonomic and cognitive abnormalities after developmental exposure to organophosphates. [ABSTRACT FROM AUTHOR]

Protein kinase C (PKC) plays a key role in many receptor-mediated signaling pathways that regulate cell growth and development. However, its roles in guiding axon growth and guidance in developing neural pathways are largely unknown. To investigate possible functions of PKC in the growth and guidance of axons in the optic chiasm, we first determined the localization of major PKC isoforms in the retinofugal pathway of mouse embryos, at the stage when axons navigate through the midline. Results showed that PKC was expressed in isoform specific patterns in the pathway. PKC-α immunoreactivity was detected in the chiasm and the optic tract. PKC-βll was strong in the optic stalk but was attenuated on axons in the diencephalon. Immunostaining for PKC-ε showed a colocalization in the chiasmatic neurons that express a surface antigen stage specific embryonic antigen-1 (SSEA-1). These chiasmatic neurons straddled the midline of the optic chiasm, and have been shown in earlier studies a role in regulation of axon growth and guidance. Expression levels of PKC-βl,-δ and -γ were barely detectable in the pathway. Blocking of PKC signaling with Ro-32-0432, an inhibitor specific for PKC-α and -β at nanomolar concentration, produced a dramatic reduction of ipsilateral axons from both nasal retina and temporal crescent. We conclude from these studies that PKC-α and -βll are the predominant forms in the developing optic pathway, whereas PKC-e is the major form in the chiasmatic neurons. Furthermore, PKC-α and -βll are likely involved in signaling pathways triggered by inhibitory molecules at the midline that guide optic axons to the uncrossed pathway. [ABSTRACT FROM AUTHOR]

Abstract: Subthreshold resonance is a characteristic membrane property of different neuronal classes, is critically involved in the generation of network oscillations, and tunes the integration of synaptic inputs to particular frequency ranges. In order to investigate whether neocortical GABAergic interneurons show resonant behavior already during early postnatal development, we performed whole-cell patch-clamp recordings from visually identified interneurons in supragranular layers of parietal regions in coronal neocortical slices from postnatal day (P) P6–P13 GAD67-GFP knock-in mice. Subthreshold resonance was analyzed by injection of sinusoidal current with varying frequency. About 50% of the investigated GABAergic interneurons showed subthreshold resonance with an average frequency of 2.0±0.2Hz (n=38). Membrane hyperpolarization to –86mV attenuated the frequency and strength of subthreshold resonance. In the presence of 1mM Ni2+ subthreshold resonance was virtually abolished, suggesting that T-type Ca2+ currents are critically involved in the generation of resonance. In contrast, subthreshold resonance was not affected by ZD7288, a blocker of HCN channels. Application of TTX suppressed subthreshold resonance at depolarized, but not hyperpolarized membrane potential, suggesting that persistent Na+ current contribute to the amplification of membrane resonance. In summary, these results demonstrate that GABAergic interneurons express subthreshold resonance at low frequencies, with T-type Ca2+ and persistent Na+ currents underlying the generation of membrane resonance. The membrane resonance of immature interneurons may contribute to the generation of slow oscillatory activity pattern in the immature neocortex and enhance the temporal precision of synaptic integration in developing cortical neurons. [Copyright &y& Elsevier]

Abstract: Mutations in MECP2 cause Rett syndrome (RTT), an X-linked neurodevelopmental disorder that primarily affects females. Individuals with RTT have increased glial fibrillary acidic protein (GFAP) expression in the brain. GFAP is an intermediate filament protein that is expressed predominately within astrocytes in the CNS. MeCP2 binds to methylated regions of the GFAP promoter region and suppresses GFAP expression in vitro. Therefore, we wanted to determine if transiently reducing MeCP2 expression would increase GFAP expression in the developing rat brain. Male and female rats received infusions of either MeCP2 or control siRNA targeting the amygdala during the first 3 days of postnatal life. Brains were collected after 6h or 2 weeks following the last infusion. MeCP2 siRNA increased GFAP mRNA and protein within the female, but not the male, amygdala on postnatal day (PN) 2. Two weeks following the infusion, levels returned to normal. MeCP2 siRNA targeting the hypothalamus also increases GFAP mRNA within the female hypothalamus on PN2, suggesting that the regulation is not brain region-specific. It appears that MeCP2 does not regulate all astrocyte markers in the developing female brain, but specifically regulates GFAP expression, as levels of S100β and vimentin were not altered in the female amygdala at either time point. These data contribute to the idea that the role of MeCP2 differs in the developing male versus female brain. Further elucidating the regulation and function of GFAP can contribute to our understanding of MeCP2 function and perhaps RTT etiology. [Copyright &y& Elsevier]

Abstract: Scope: Ascorbic acid (ascorbate) is required to recycle tetrahydrobiopterin, which is necessary for neurotransmitter synthesis by the rate-limiting enzymes tyrosine and tryptophan hydroxylases. We sought to determine whether ascorbate might regulate embryonic brain cortex monoamine synthesis utilizing transgenic mouse models with varying intracellular ascorbate levels. Methods and results: In embryos lacking the sodium-dependent vitamin C transporter 2 (SVCT2), very low levels of brain ascorbate decreased cortex levels of norepinephrine and dopamine by approximately 33%, but had no effect on cortex serotonin or its metabolite, 5-hydroxyindole acetic acid. This decrease in ascorbate also led to a decrease in protein levels of tyrosine hydroxylase, but not of tryptophan hydroxylase. Increased cortex ascorbate in embryos carrying extra copies of the SVCT2 resulted in increased levels of dopamine and its metabolite, 3,4-dihydroxyphenylacetic acid (DOPAC), as well as serotonin and 5-hydroxyindole acetic acid. Conclusion: The dependence of embryonic brain cortex neurotransmitter synthesis and tyrosine hydroxylase expression on intracellular ascorbate emphasizes the importance of receiving adequate ascorbate during development. [Copyright &y& Elsevier]

Abstract: Vesicular glutamate transporter 3 (VGLUT3) plays an important role in hearing, and VGLUT3 knockout mice are deaf. However, the mechanisms whereby VGLUT3 exerts its effects in the cochlea are not well established. Elucidating the developmental and aging dynamics of VGLUT3 localization and expression in the cochlea would aid a functional understanding of auditory glutamatergic transmission. In this study, we characterized the expression of VGLUT3 in rat auditory epithelium and spiral ganglion cells (SGCs) during postnatal development and with age using immunohistochemistry and quantitative real-time reverse transcription polymerase chain reaction (RT-PCR). We observed VGLUT3 expression in cochlea inner hair cells (IHCs) and SGCs as early as postnatally, and its expression increased gradually with maturity. However, the expression of VGLUT3 in the IHCs decreased in the aging rats compared to postnatal day 60 (P60), and VGLUT3 was still expressed in the remaining SGCs in the aging rats. VGLUT3-immunopositive punctate structures were only found in the cytoplasm of SGCs, not IHCs, at P7, or later. Variation in VGLUT3 mRNA expression evaluated by quantitative real time RT-PCR was consistent with the immunohistochemical data except for no statistical difference between P60 and aging rats in the whole cochlea level. Our data suggest that VGLUT3 likely has developmental and physiological roles in the rat cochlea during postnatal development as well as later in life. VGLUT3 may have additional unknown roles in SGCs besides glutamate transport. [Copyright &y& Elsevier]

Abstract: In humans, Fragile X Syndrome (FXS) is characterized by enhanced fear, hyperactivity, social anxiety, and, in a subset of individuals, autism. Many of the emotional and social deficits point to defects in the amygdala. We have previously shown defects in inhibitory neuron drive onto excitatory projection neurons in the basolateral amygdala (BLA) of juvenile Fmr1 -/y knockout (KO) mice. Using pharmacological approaches, we have also previously revealed dynamic functional deficits in α1, α2, and α3 subunit-containing GABAA receptors (GABAARs α1, α2, and α3) during early postnatal development. In this study, we sought to determine whether these defects in GABAAR function are accompanied by changes in protein expression of GABAARs α1, α2, and α3 and the post-synaptic GABAAR-clustering protein gephyrin. Interestingly, we found that while the expression of these proteins did not significantly differ between wildtype (WT) and KO mice at each time point, the timing of developmental expression of GABAAR α1, α2, and gephyrin was altered. Collectively, these data reveal novel defects in inhibitory synapse protein expression during critical periods of early postnatal development that could contribute to observed inhibitory neurotransmission deficits in the KO mouse BLA. [Copyright &y& Elsevier]

Abstract: Many neurons degenerate after injuries resulting from overstimulation, drugs, genetic mutations, and aging. Although several growth factors and neurotrophins delay degeneration and promote regrowth of neural processes, the role of fibroblast growth factor 8 (FGF8) in mammalian spiral ganglion neurons (SGN) neurite outgrowth has not been examined. This study develops and uses SGN cell cultures suitable for experimental analysis, it investigates whether FGF8a and FGF8b isoforms affect the neurite outgrowth from SGN cultured in vitro. We found that both FGF8a and FGF8b promoted the outgrowth of neurites from cultured SGN. This response is mediated by FGF receptors and involves the activation of IκBα-mediated NFκB signaling pathway. These findings suggest that, besides its morphogenetic role during development, FGF8 may have trophic functions in the adult which are relevant to regeneration. [Copyright &y& Elsevier]

Abstract: Using a well-established model of binge-like ethanol treatment of rat pups on postnatal days (PD) 4–9, we found that maturation of GABAA receptor (GABAAR) miniature postsynaptic currents (mPSCs) was substantially blunted for medial septum/diagonal band (MS/DB) neurons in brain slices on PD 11–16. Ethanol reduced mPSC amplitude, frequency, and decay kinetics, while attenuating or exaggerating allosteric actions of zolpidem and allopregnanolone, respectively. The impact of ethanol in vivo was long lasting as most changes in MS/DB GABAAR mPSCs were still observed as late as PD 60–85. Maturing MS/DB neurons in naïve brain slices PD 4–16 showed increasing mPSC frequency, decay kinetics, and zolpidem sensitivity that were nearly identical to our earlier findings in cultured septal neurons (DuBois et al., 2004, 2006). These rapidly developing mPSC parameters continued to mature through the first month of life then stabilized throughout the remainder of the lifespan. Finally, equivalent ethanol-induced alterations in GABAAR mPSC signaling were present in MS/DB neurons from both male and female animals. Previously, we showed ethanol treatment of cultured embryonic day 20 septal neurons distorts the maturation of GABAAR mPSCs predicting that early stages of GABAergic transmission in MS/DB neurons are vulnerable to intoxication injury (DuBois et al., 2004, 2006). Since the overall character, timing, and magnitude of GABAergic mPSC developmental- and ethanol-induced changes in the in vivo model so closely mirror chronologically equivalent adaptations in cultured septal neurons, this suggests that such parallel models of ethanol impairment of GABAergic synaptic development in vivo and in vitro should be useful for translational studies exploring the efficacy and mechanism of action of potential therapeutic interventions from the cellular to whole animal level. [Copyright &y& Elsevier]

Abstract: High-frequency oscillations in the brain open a new window for studies of language development in humans. The objective of this study is to determine the spatiotemporal and frequency signatures of word processing in healthy children. Sixty healthy children aged 6–17 years were studied with a whole-cortex magnetoencephalography (MEG) system using a word recognition paradigm optimized for children. The temporal signature of neuromagnetic activation was measured using averaged waveforms. The spatial and frequency signatures of neuromagnetic activation were assessed with wavelet-based beamformer analyses. The results of waveform analyses showed that the latencies of the first and third neuromagnetic responses changed with age (p<0.01). The source imaging data revealed a clear lateralization of source activation in the 70–120Hz range in children within the age range of 6 to 13 years of age (p<0.01). Males and females demonstrated different developmental trajectories over the age range of 9 to 13 years of age (p<0.01). These findings suggest that left-hemisphere language processing emerges from early bilateral brain areas with gender optimal neural networks. The neuromagnetic signatures of language development in healthy children may be used as references for future identification of aberrant language function in children with various disorders. [Copyright &y& Elsevier]

Abstract: Chronic exposure to low doses of organophosphates during brain development can induce persistent neurochemical and behavioral effects. This study sought to determine the long-lasting effects of developmental exposure to chlorpyrifos (CPF) and diazinon (DZN) on passive avoidance (PA) performance and neuronal nitric oxide synthase (nNOS)-containing neurons in the subnuclei within basolateral complex of amygdala (BLC). Developing rats were exposed to daily dose (1mg/kg) of CPF or DZN during gestational days 15–18 and postnatal days (PND) 1–4. PA performance was assessed in young adulthood (PND 60). Brain sections were also processed by NADPH-diaphorase (NADPH-d) and nNOS immunohistochemistry. Gestational exposure to CPF increased NADPH-d+/nNOS-immunoreactive (IR) neurons within the basolateral nucleus (BL) and medial paracapsular intercalated cluster, which was along with PA retention impairment in both male and female rats. Prenatal exposure to DZN did not significantly change the number of NADPH-d+/nNOS-IR neurons in the BLC while impaired PA retention in females. Postnatal exposure to CPF decreased NADPH-d+/NOS-IR neurons in the BL without affecting PA performance. Exposure to DZN during early postnatal period impaired PA retention in both sexes, albeit to a lesser extent in females, and was along with a considerable sex independent reduction of NADPH-d+/NOS-IR neurons in all BLC subnuclei. Our data suggest that developmental exposure to apparently subtoxic dose of CPF and DZN elicit long-lasting impairment in PA retention that are associated, but not necessarily correlated with effects on NADPH-d+/NOS-IR neurons in BLC of the amygdala. [Copyright &y& Elsevier]

Abstract: The embryonic development of synapses in the rostral nucleus of the solitary tract (rNST) was investigated in rat to determine when synapses begin to function. Using a brain slice preparation we studied appearance of synaptic receptors on second order rNST neurons and investigated the development of postsynaptic responses elicited by afferent nerve stimulation. Prenatal excitatory and inhibitory synaptic responses were recorded as early as E14. Glutamatergic and GABAergic postsynaptic responses were detected as early as E16. Both NMDA and AMPA receptors contributed to glutamatergic postsynaptic responses. GABAergic postsynaptic responses resulted primarily from activation of GABAA receptors. However, functional GABAC receptors were also demonstrated. A glycinergic postsynaptic response was not found although functional glycine receptors were demonstrated at E16. Solitary tract (ST) stimulation-evoked EPSCs, first detected at E16, were eliminated by glutamate receptor antagonists. ST-evoked IPSPs, also detected at E16, were eliminated by GABAA receptor antagonist. Thus, considerable prenatal development of rNST synaptic connections occurs and this will ensure postnatal function of central taste processing circuits. [Copyright &y& Elsevier]

Abstract: The pluripotent P19 embryo carcinoma cell line was studied to determine a signaling pathway regulating MeCP2 expression. P19 cells were induced to differentiate into neurons by RA and express β-III tubulin at one day after induction and synaptophysin by 7 days. MeCP2 was first observed after β-III tubulin expression was detected and continued to rise over the course of differentiation. Both Mecp2 e1 and e2 mRNA forms progressively increased in differentiating cells. MeCP2 expression was increased by tumor necrosis factor (TNF) in early differentiating cells, which was blocked by NFκB inhibitors. TNF did not increase MeCP2 expression in naïve cells. Moreover, TNF did not increase NFκB reporter gene activity in naïve cells even though increases were observed in early differentiating cells. The protein kinase C activator phorbol 12-myristate 13-acetate (PMA) increased MeCP2 expression in naïve P19 cells, which was also blocked by NFκB inhibitors. Interestingly, PMA increased NFκB reporter gene activity in naïve cells. Finally, PMA, but not TNF, induced IκBα degradation in naïve P19 cells. Taken together, our data indicates that MeCP2 expression is regulated in part by signaling pathways involving NFκB. [Copyright &y& Elsevier]

Abstract: In groups of 7-year-olds and 11-year-olds, event-related potentials (ERPs) were recorded to briefly presented, masked letter strings that included real word (DARK/PARK), pronounceable pseudoword (DARL/PARL), unpronounceable nonword (RDKA/RPKA), and letter-in-xs (DXXX, PXXX) stimuli in a variant of the Reicher–Wheeler paradigm. Behaviorally, participants decided which of two letters occurred at a given position in each string (here, forced-choice alternatives D and P). Both groups showed evidence of behavioral word (more accurate choices for letters in words than in baseline nonwords or letter-in-xs) and pseudoword (more accurate choices for letters in pseudowords than in baseline nonwords or letter-in-xs) superiority effects. Electrophysiologically, 11-year-olds evidenced superiority effects on P150 and N400 peak amplitude, while 7-year-olds showed effects only on N400 amplitude. These findings suggest that the mechanisms underlying the observed behavioral superiority effects may be lexical in younger children but both sublexical and lexical in older children. These results are consistent with a lengthy developmental time course for automatic sublexical orthographic specialization, extending beyond the age of 11. [Copyright &y& Elsevier]

Abstract: Dexamethasone (DEX), a synthetic glucocorticoid, has been used to treat respiratory distress syndrome in prematurely born infants. Despite the important short-term benefit on lung function, there is growing concern about the long-term outcome of this treatment, since follow-up studies of prematurely born infants have shown lasting adverse neurodevelopmental effects. Since the mechanism underlying these neurodevelopmental impairments is largely unknown, the aim of the present study was (i) to investigate the acute effects of neonatal DEX treatment on the developing brain; and (ii) to block specifically the effects of DEX on the brain by central administration of the glucocorticoid receptor (GR) antagonist mifepristone. Long Evans rat pups were injected subcutaneously with tapering doses of DEX or saline (SAL) on postnatal days (pnd) 1, 2 and 3. Separate groups received intracerebroventricular injections with mifepristone prior to DEX treatment. On pnd 4 and 10, pups were sacrificed and brains collected for analysis of cell proliferation (Ki-67) and astrogliosis (GFAP). We report that neonatal DEX treatment reduced hippocampal cell proliferation on pnd 4, an effect that was normalized by pnd 10. Although on pnd 4, GFAP expression was not affected, DEX treatment caused a significant reduction in the number and density of astrocytes in hippocampus and corpus callosum on pnd 10, which was normalized by mifepristone pre-treatment. These acute alterations in the neonate brain might underlie later functional impairments reported in DEX-treated animals and humans and further illustrate the impact of early GR activation on brain development. [Copyright &y& Elsevier]

Abstract: Status epilepticus (SE) when occurred during brain development can cause short- and long-term consequences, which are frequently associated with NMDA-mediated glutamatergic excitotoxicity. In the present work, we investigated the putative neuroprotective role of ketamine, an NMDA receptor antagonist, on early life SE-induced acute neuronal death and long-term behavioral abnormalities. Male Wistar rats (16 postnatal days) were induced to SE by LiCl–pilocarpine i.p. administration (3mEq/kg; 60mg/kg, respectively). Fifteen or 60min after pilocarpine injection, animals received a ketamine administration (22.5mg/kg i.p.). Neuronal degeneration was assessed 24h after SE induction. Another subset of animals was destined to behavioral tasks in adulthood (75–80 postnatal days). Fluoro-Jade C labeling revealed a marked neuronal death on CA1 hippocampal subfield, habenula, thalamus and amygdala in SE animals. Ketamine post-SE onset treatment prevented neuronal death in all regions assessed. In the elevated plus maze, SE induced an increase in anxiety-like behaviors whereas ketamine administration during seizures was able to prevent this alteration. Ketamine administration in non-SE animals resulted in high anxiety levels. There were no observed differences among groups in the open field task in all parameters analyzed. Our results suggest that ketamine post-SE onset treatment was effective in preventing acute and long-standing alterations caused by SE early in life, which indicates a putative role of glutamatergic system on SE-induced brain damage as well as long-lasting behavioral consequences. [Copyright &y& Elsevier]

Abstract: To assess developmental characteristics of nociceptive responses induced by bee venom (BV) injection in neonatal rats, we exposed pups to intra-plantar injection of various BV concentrations given at different time points between postnatal day 1 and day 28 (P1–P28). Persistent spontaneous nociception (PSN) as well as thermal and mechanical nociceptive response was compared before and after a BV injection. There were distinct age-related changes in the baseline paw withdrawal thermal latency (PWTL) and paw withdrawal mechanical threshold (PWMT) when examined on P1, P4, P7, P14, P21, and P28. The lowest and highest baseline PWTL was shown on P1 and P7, respectively, and PWTL was unchanged from P7 to P28. In contrast, the baseline PWMT remained low before P21 but increased dramatically afterward. Neonatal rats receiving intra-plantar BV injection showed a time-dependent change in nociceptive responses, including (1) a dose-related increase in PSN from P1 to P28; (2) a non-specific decrease (indistinguishable between saline and BV injection) in PWTL and PWMT up to P14 and P21, respectively; and (3) a specific decrease (in response to BV injection only) in PWTL and PWMT after P14 and P21, respectively. These findings indicate that characteristic changes in the baseline and BV-induced nociceptive response are both time-dependent and modality-specific in neonatal rats. The data reveal a critical postnatal period during which nociceptive stimulation could have a significant influence on nociceptive behavior in adult rats and suggest that preclinical models of neonatal nociception should be evaluated according to different postnatal time points. [Copyright &y& Elsevier]

Abstract: The zebrafish pineal gland plays an important role in different physiological functions including the regulation of the circadian clock. In the fish pineal gland the pinealocytes are made up of different segments: outer segment, inner segment and basal pole. Particularly, in the outer segment the rhodopsin participates in the external environment light reception that represents the first biochemical step in the melatonin production. It is well known that the rhodopsin in the adult zebrafish is well expressed in the pineal gland but both the expression and the cellular localization of this protein during development remain still unclear. In this study using qRT-PCR, sequencing and immunohistochemistry the expression as well as the protein localization of the rhodopsin in the zebrafish from larval (10dpf) to adult stage (90dpf) were demonstrated. The rhodopsin mRNA expression presents a peak of expression at 10dpf, a further reduction to 50dpf before increasing again in the adult stage. Moreover, the cellular localization of the rhodopsin-like protein was always localized in the pinealocyte at all ages examined. Our results demonstrated the involvement of the rhodopsin in the zebrafish pineal gland physiology particularly in the light capture during the zebrafish lifespan. [Copyright &y& Elsevier]

Abstract: During early postnatal brain development, experience-driven delivery of AMPA receptors to synapses participates in the initial organization of cortical function. By combining virus-mediated in vivo gene delivery with in vitro whole cell recordings, we identified a subunit-specific developmental program of experience-driven AMPA receptor delivery to synapses in rat barrel cortex. We expressed green fluorescent protein (GFP)-tagged AMPA receptors (GFP-GluR1, or GFP-GluR4) into layer 2/3 pyramidal neurons at two distinct developmental periods, postnatal day (P)8–P10 and P12–P14. Two days after viral infection, acute brain slices were prepared, and synaptic transmission from layer 4 to layer 2/3 was analyzed by whole cell recordings. We found that whisker experience drives GluR4 but not GluR1 into these synapses early in postnatal development (P8–P10). However, at P12–14, GluR1 but not GluR4 is delivered into synapses by whisker experience. This precise developmental plan suggests unique plasticity properties endowed in different AMPA receptor subunits which shape the initial experience-driven organization of cortical function. [Copyright &y& Elsevier]

Abstract: To evaluate the potentially disrupting effects of environmental estrogens on neuroendocrine networks controlling reproduction, we studied the impact of the pharmaceutical product 17-α-ethinylestradiol (EE2) on gonadotropin-releasing hormone (GnRH-1) neuron development in mouse embryo. Pregnant mice were treated per os with EE2 at 0.01, 0.1 or 1μg/kg/day, between embryonic days 10.5 (E10.5) and E13.5, a period during which GnRH-1 neurons are generated and start their intra-nasal migration. Embryos at E13.5 were examined and processed for GnRH-1 immunohistochemistry. Immunopositive neurons were counted all along their migratory path. A short oral administration of environmentally relevant doses of EE2 to pregnant mice had a significant impact on whole embryo development, leading to a limited but significant growth retardation. The total number of GnRH-1 neurons was statistically significantly increased in a dose-dependent manner. The repartition of GnRH-1 neurons along their migratory path was not affected by EE2 treatment. These results suggest an impact of environmental EE2 concentrations on embryonic GnRH-1 development through a modulation of neurogenesis and/or apoptosis. [Copyright &y& Elsevier]

Abstract: Smoking during pregnancy exposes the fetus to nicotine, resulting in nicotine-stimulated neurotransmitter release. Recent evidence suggests that the hippocampus develops differently in males and females with delayed maturation in males. We show that chronic nicotine exposure during the first postnatal week has sex-specific long-term effects. Neonatal rat pups were chronically treated with nicotine (6mg/kg/day) (CNN) from postnatal day 1 to 7 or milk only (Controls), and hippocampal slices were prepared from Control- and CNN-treated young adults. Field excitatory postsynaptic potentials (fEPSPs) or population spikes (PSs) were recorded from the CA1 hippocampus following CA1 s. radiatum stimulation. Input/Output curves constructed from fEPSP data indicated that CNN-males, but not females, had significantly increased excitatory responses compared to Controls (p<0.05, n=10 Con, n=11 CNN). Long-term potentiation (LTP) was not significantly changed by CNN. In the presence of bicuculline, which blocks inhibitory GABAA receptors, an epileptiform burst consisting of a series of PSs was evoked. The amplitude of the first PS was significantly larger in CNN-males and females compared to Controls (males: p<0.01, n=8 Con, n=8 CNN; females: p<0.05, n=9 Con, n=7 CNN). Only CNN-males also had significantly larger second PSs (p<0.05, n=8 con, n=8 CNN). Epileptiform activity evoked by zero Mg2+ incubation did not differ in amplitude or duration of bursts in CNN-males or females compared to Controls. These data indicate that neonatal nicotine exposure has long lasting effects and results in increased excitation within the CA1 hippocampus in adulthood, with males showing increased sensitivity to nicotine''s effects. [Copyright &y& Elsevier]

Abstract: The primary afferent neurons of the vestibular ganglion convey sensory information from hair cells in the semicircular canals and otolith organs to the vestibular nuclei, the adjacent brainstem and the cerebellum. The intrinsic firing properties of vestibular ganglion cells (VGCs) are heterogeneous and have been classified into phasic, intermediate and tonic firing types on the basis of their response to injected depolarizing currents. A previous study from our group showed that the proportion of phasic discharging VGCs decreased during the first postnatal weeks. Moreover, α-dendrotoxin (α-DTX), a Kv1 potassium channels antagonist, turned neuron phasic firing to tonic, thus suggesting that these channels play an important role in the developmental changes of VGCs firing patterns. Here, by using immunohistochemistry, Western blotting and quantitative real-time reverse transcriptase-polymerase chain reaction (RT-PCR), we explored the change in the expression of α-DTX-sensitive K+ channels, Kv1.1, Kv1.2 and Kv1.6 in rat VGCs during early postnatal periods. We showed that expression of Kv1.6 protein is down-regulated together with expression of Kv1.6 mRNA after postnatal day 7 in rat VGCs whereas expression of Kv1.1 and Kv1.2 proteins did not change during the same developmental period. Our results suggest that down-regulation of the Kv1.6 protein and mRNA may be associated with maturation of excitable properties of primary vestibular neurons. [Copyright &y& Elsevier]

Abstract: The present study compares the electrophysiological properties and the primary pacemaker currents that flow during the interspike interval in locus coeruleus (LC) neurons from infant (P7–12days) and young adult (8–12weeks) mice. The magnitude of the primary pacemaker currents, which consist of an excitatory TTX-sensitive Na+ current and an inhibitory voltage-dependent K+ current, increased in parallel during development. We found no evidence for the involvement of hyperpolarization-activated (IH) or Ca2+ currents in pacemaking in infant or adult LC neurons. The incidence of TTX-resistant spikes, observed during current clamp recordings, was greater in adult neurons. Neurons from adult animals also showed an increase in voltage fluctuations, during the interspike interval, as revealed in the presence of the K+ channel blocker, 4-AP (1mM). In summary, our results suggest that mouse LC neurons undergo changes in basic electrophysiological properties during development that influence pacemaking and hence spontaneous firing in LC neurons. [Copyright &y& Elsevier]

Abstract: The melanopsin-expressing retinal ganglion cells are specialized in measuring irradiance for several functions, including daily photoentrainment and regulation of pupil size. In the present study, these cells were analyzed in mice during their perinatal period, from embryonic day (E) 15 to postnatal day (P) 1. Melanopsin expression was detected at E15 in cells that did not co-express the transcription factor Brn3a. Under light/dark (LD) cycles, the number of melanopsin-expressing cells did not change between E16 and E19, while a very significant increase was observed during the short interval around birth, between E19 (the day before birth) and P0 (the day of birth). As these samples were collected after lights on, to determine whether such increase in melanopsin expression was driven by light, we also analyzed samples collected 0–4hours after birth (during the night period), which revealed that the cell number increase was already present and, therefore, was not induced by the early post-birth light exposure. To clarify the role of ambient light conditions during this period, P1 retinas from pups under constant light or darkness conditions were also analyzed and compared to those of mice under LD cycles. No variation in the number of immunostained cells was detected among the groups studied, indicating that ambient conditions did not provoke the increase in melanopsin expression detected. Rather, it might have been induced by either a maternal or a developmental signal and is likely related to the first connections between the retina and the suprachiasmatic nucleus reported by other authors. [Copyright &y& Elsevier]