Michael Gruss

MATERNAL SEPARATION DURING A SPECIFIC POSTNATAL TIME WINDOW PREVENTS REINFORCEMENT OF HIPPOCAMPAL LONG-TERM POTENTIATION IN ADOLESCENT RATS

In an attempt to develop an animal model to study the etiology of brain dysfunction in relation to early life experience, we tested the hypothesis that early-life stress during specific postnatal time windows affects long-term potentiation (LTP) reinforcement in adolescence. Male Wistar rat pups were stressed by separation from their dams for 24 h at postnatal day (PND) 4, 9, or 18. The animals were tested for reinforcement of LTP at adolescence (9 weeks old) by exposing them to a 2-min swim-stress. Here, we show that maternal separation during (at PND9) but not at the beginning (at PND4) or after (at PND18) the stress-hyporesponsive-period of the hypothalamic-pituitary-adrenal-axis impairs emotional LTP-reinforcement in adolescent animals. Thus, this in vivo model allows the investigation of physiological and pathophysiological emotional information processing at the cellular level in freely behaving adolescent animals.

Stimulus‐Evoked Increase of Glutamate in the Mediorostral Neostriatum/Hyperstriatum Ventrale of Domestic Chick After Auditory Filial Imprinting: An In Vivo Microdialysis Study

Abstract: Imprinting in chicks is a form of juvenile learning that has been used to study the basic cellular mechanisms of learning and memory. The forebrain area mediorostral neostriatum/hyperstriatum ventrale (MNH) is a center for acoustic imprinting. Electrophysiological and pharmacological behavioral studies in the MNH have demonstrated that the glutamatergic system and the associated receptors are critically involved in auditory filial imprinting. Accordingly, we investigated the hypothesis that stimulus‐evoked glutamate release may be altered after this learning process. Using an in vivo microdialysis technique, we observed a significantly higher increase of extracellular glutamate level in tone‐imprinted chicks during exposure to the previously imprinted tone than in socially imprinted control chicks. In a further series of experiments, where we exposed animals from both experimental groups to handling distress, glutamate levels in MNH showed only a slight increase, whereas we observed a pronounced increase of extracellular glutamate in the lobus parolfactorius (LPO), the avian analogue of the basal ganglia. No difference of distress‐evoked glutamate release was found in MNH and LPO between tone‐imprinted and socially imprinted chicks. The tone‐evoked enhanced glutamate response in tone‐imprinted chicks suggests that during auditory imprinting glutamatergic synapses develop the potential to increase transmitter release in response to the imprinting stimulus.

Passive Avoidance Training and Recall are Associated With Increased Glutamate Levels in the Intermediate Medial Hyperstriatum Ventrale of the Day-Old Chick

In the young chick, the intermediate medial hyperstriatum ventrale is involved in learning paradigms, including imprinting and passive avoidance learning. Biochemical changes in the intermediate medial hyperstriatum ventrale following learning include an up-regulation of amino-acid transmitter levels and receptor activity. To follow the changes of extracellular amino acid levels during passive avoidance training, we used an in vivo microdialysis technique. Probes were implanted in chicks before training the animals, either on a methyl- anthranylate-or water-coated bead. One hour later, recall was tested in both groups by presenting a similar bead. An increase of extra-cellular glutamate levels accompanied training and testing in both groups; during training, glutamate release was higher in methylanthranylate- trained than in water-trained chicks. When compared with the methylanthranylate-trained chicks during testing, the water-trained chicks showed enhanced extra-cellular glutamate levels. No other amino acid examined showed significant changes. After testing, the chicks were anesthetized and release- stimulated with an infusion of 50 mM potassium. Extra-cellular glutamate and taurine levels were significantly increased in both methylanthranylate-and water-trained chicks. The presentation of methylanthranylate as an. olfactory stimulus significantly enhanced glutamate levels, especially in methylanthranylate-trained chicks. The results suggest that such changes in extra-cellular glutamate levels in the intermediate medial hyperstriatum ventrale accompany pecking at either the water- or the methylanthranylate-bead. The taste of the aversant may be responsible for the greater increases found in methylanthranylate-trained birds.

Early stress and chronic methylphenidate cross‐sensitize dopaminergic responses in the adolescent medial prefrontal cortex and nucleus accumbens

Methylphenidate (MP) is widely used to treat attention deficit/hyperactivity disorder in children. However, basic research has been mainly focused on MP treatment in adult, behaviorally normal rodents. Here we analyzed MP‐evoked changes of dopamine (DA) release in the limbic system of juvenile rodents with hyperactive and attention deficit‐like symptoms. Using dual probe in vivo microdialysis, DA levels were quantified in the medial prefrontal cortex and nucleus accumbens of juvenile and adolescent degus (Octodon degus). Acute stress‐ and acute MP‐evoked dopaminergic responses in normal juvenile and adolescent animals were compared with (i) animals showing symptoms of hyperactivity and attention deficits induced by early life stress, i.e. repeated parental separation during the first 3 weeks of life, and (ii) animals chronically treated with MP during pre‐adolescence. Our main results revealed that (i) early life stress and (ii) chronic MP treatment during pre‐adolescence cross‐sensitize limbic dopaminergic functions in adolescent animals. Furthermore, we demonstrated a unique pattern of acute MP‐evoked DA release in the juvenile compared with the adolescent medial prefrontal cortex and nucleus accumbens. Our findings that the functional maturation of dopaminergic limbic function is significantly altered by early life experience, i.e. repeated parental separation and chronic MP treatment, allow novel insights into the etiology of attention deficit/hyperactivity disorder and into the long‐term consequences of MP treatment on brain development.

Age- and region-specific imbalances of basal amino acids and monoamine metabolism in limbic regions of female Fmr1 knock-out mice

The Fragile X syndrome, a common form of mental retardation in humans, originates from the loss of expression of the Fragile X mental retardation gene leading to the absence of the encoded Fragile X mental retardation protein 1 (FMRP). A broad pattern of morphological and behavioral abnormalities is well described for affected humans as well as Fmr1 knock-out mice, a transgenic animal model for the human Fragile X syndrome. In the present study, we examined neurochemical differences between female Fmr1 knock-out and wildtype mice with particular focus on neurotransmission. Significant age- and region-specific differences of basal tissue neurotransmitter and metabolite levels measured by high performance liquid chromatography were found. Those differences were more numerous in juvenile animals (postnatal day (PND) 28-31) compared to adults (postnatal day 209-221). In juvenile female knock-out mice, especially aspartate and taurine were increased in cortical regions, striatum, cerebellum, and brainstem. Furthermore, compared to the wildtype animals, the juvenile knock-out mice displayed an increased level of neuronal inhibition in the hippocampus and brainstem reflected by decreased ratios of (aspartate + glutamate)/(taurine + GABA), as well as an increased dopamine (DA) turnover in cortical regions, striatum, and hippocampus. These results provide the first evidence that the lack of FMRP expression in female Fmr1 knock-out mice is accompanied by age-dependent, region-specific alterations in brain amino acids, and monoamine turnover, which might be related to the reported synaptical and behavioural alterations in these animals.

Alterations of amino acids and monoamine metabolism in male Fmr1 knockout mice: a putative animal model of the human fragile X mental retardation syndrome.

The Fragile X syndrome, a common form of mental retardation in humans, is caused by silencing the fragile X mental retardation (FMR1) geneleading to the absence of the encoded fragile X mental retardation protein 1 (FMRP). We describe morphological and behavioral abnormalities for both affected humans and Fmr1 knockout mice, a putative animal model for the human Fragile X syndrome. The aim of the present study was to identify possible neurochemical abnormalities in Fmr1 knockout mice, with particular focus on neurotransmission. Significant region-specific differences: of basal neurotransmitter and metabolite levels were found between wildtype and Fmr1 knockout animals, predominantly in juveniles (post-natal days 28 to 31). Adults (postnatal days 209 to 221) showed only few abnormalities as compared with the wildtype. In juvenile knockout mice, aspartate and taurine were especially increased in cortical regions, striatum, hippocampus, cerebellum, and brainstem. In addition, juveniles showed an altered balance between excitatory and inhibitory amino acids in the caudal cortex, hippocampus, and brainstem. We detected very few differences in monoamine turnover in both age stages. The results presented here provide the first evidence that lack of FMRP expression in FMRP knockout mice is accompanied by age-dependent, region-specific alterations in neurotransmission.

Epigenetic modulation of the developing serotonergic neurotransmission in the semi-precocial rodent Octodon degus.

Environmental influences during early life periods, particularly those provided by the mother or parents, are generally considered to have a strong impact on the development of brain and behaviour of the offspring. In the semi-precocial South American species Octodon degus, a rodent becoming increasingly popular in different laboratory research fields, the present study aimed to examine the consequences of the disturbance of the parent-offspring interaction induced by parental separation on the serotonergic neurotransmission. Based on a quantitative neurochemical approach using brain homogenates obtained from cortical regions and the hippocampus our results revealed that (i) the tissue levels of serotonin and 5-hydroxyindoleacetic acid showed in both sexes a moderate, around two-fold increase until adulthood, indicating relatively matured cortical and hippocampal serotonergic systems at birth. In addition, we found an age-, region- and sex-specific pattern of changes in the serotonergic system induced by (ii) an acute stress challenge early in life (1-h parental separation at the postnatal day 3, 8, 14 or 21) with the most pronounced effects at earlier ages (between postnatal days 3 and 14) in the female cortex and (iii) repeated stress exposure (1h daily) during the first 3 weeks of life affecting cortical regions of both sexes. Taken together, these data indicate that early life stress (i.e. parental separation) influences the developing serotonergic system in the semi-precocial O. degus, even if the brain is relatively well matured at the early stages of postnatal development.

Impact of an additional chronic BDNF reduction on learning performance in an Alzheimer mouse model

There is increasing evidence that brain-derived neurotrophic factor (BDNF) plays a crucial role in Alzheimer’s disease (AD) pathology. A number of studies demonstrated that AD patients exhibit reduced BDNF levels in the brain and the blood serum, and in addition, several animal-based studies indicated a potential protective effect of BDNF against Aβ-induced neurotoxicity. In order to further investigate the role of BDNF in the etiology of AD, we created a novel mouse model by crossing a well-established AD mouse model (APP/PS1) with a mouse exhibiting a chronic BDNF deficiency (BDNF+/−). This new triple transgenic mouse model enabled us to further analyze the role of BDNF in AD in vivo. We reasoned that in case BDNF has a protective effect against AD pathology, an AD-like phenotype in our new mouse model should occur earlier and/or in more severity than in the APP/PS1-mice. Indeed, the behavioral analysis revealed that the APP/PS1-BDNF+/−-mice show an earlier onset of learning impairments in a two-way active avoidance task in comparison to APP/PS1- and BDNF+/−-mice. However in the Morris water maze (MWM) test, we could not observe an overall aggrevated impairment in spatial learning and also short-term memory in an object recognition task remained intact in all tested mouse lines. In addition to the behavioral experiments, we analyzed the amyloid plaque pathology in the APP/PS1 and APP/PS1-BDNF+/−-mice and observed a comparable plaque density in the two genotypes. Moreover, our results revealed a higher plaque density in prefrontal cortical compared to hippocampal brain regions. Our data reveal that higher cognitive tasks requiring the recruitment of cortical networks appear to be more severely affected in our new mouse model than learning tasks requiring mainly sub-cortical networks. Furthermore, our observations of an accelerated impairment in active avoidance learning in APP/PS1-BDNF+/−-mice further supports the hypothesis that BDNF deficiency amplifies AD-related cognitive dysfunctions.

9-Methyl-β-carboline-induced cognitive enhancement is associated with elevated hippocampal dopamine levels and dendritic and synaptic proliferation.

J. Neurochem. (2012) 121, 924–931.

Cognitive training during infancy and adolescence accelerates adult associative learning: Critical impact of age, stimulus contingency and training intensity

A growing body of evidence supports the hypothesis that juvenile cognitive training shapes neural networks and behavior, and thereby determines the adults capacity for learning and memory. In particular, we have shown that infant rats, even though they do not develop an active avoidance strategy in a two-way active avoidance task, show as adults accelerated learning in the same learning task. This indicates that a memory trace was formed in the infant rats, which most likely is recruited during adult training. To identify the learning conditions, which are essential prerequisites to form this memory trace in infancy or adolescence, we investigated the critical impact of: (i) age, (ii) CS-UCS contingency, and (iii) pre-training intensity on this facilitating effect. We observed: (i) an age-dependent improvement of avoidance learning, (ii) that the beneficial impact of infant or adolescent pre-training on adult learning increases with the age at pre-training, (iii) that CS-UCS contingency during infant pre-training was most efficient to accelerate adult learning, (iv) that pre-training intensity (i.e. number of pre-training trials) was positively correlated with the pre-training induced acceleration of adult learning, and (v) that infant rats, compared to adolescent rats, need a higher training intensity to show learning improvement as adults. These results indicate that infant rats develop a goal-oriented escape strategy, which during adult training is replaced by an avoidance strategy, facilitated by the recruitment of the CS-UCS association, which has been learned during infant training. Based on these results the future challenge will be to identify the specific contribution of prefronto-limbic circuits in infant and adult learning in relation to their functional maturation.