Changes in reward-induced neural activity upon Cafeteria Diet consumption.

Excessive consumption of highly palatable foods rich in sugar and fat, often referred to as "junk" or "fast" foods, plays a central role in the development of obesity. The highly palatable characteristics of these foods activate hedonic and motivational mechanisms to promote food-seeking behavior and overeating, which is largely regulated by the brain reward system. Excessive junk food consumption can alter the functioning of this reward system, but exact mechanisms of these changes are still largely unknown. This study investigated whether long-term junk food consumption, in the form of Cafeteria (CAF) diet, can alter the reward system in adult, female Long-Evans rats, and whether different regimes of CAF diet influence the extent of these changes. To this end, rats were exposed to a 6-week diet with either standard chow, or ad libitum daily access to CAF diet, 30 % restricted but daily access to CAF diet, or one-day-a-week (intermittent) ad libitum access to CAF diet, after which c-Fos expression in the Nucleus Accumbens (NAc), Prefrontal Cortex (PFC), and Ventral Tegmental Area (VTA) following consumption of a CAF reward of choice was examined. We found that all CAF diet regimes decreased c-Fos expression in the NAc-shell when presented with a CAF reward, while no changes in c-Fos expression upon the different diet regimes were found in the PFC, and possibly the VTA. Our data suggests that long-term junk food exposure can affect the brain reward system, resulting in an attenuated activity of the NAc-shell.

High tracking of apolipoprotein B levels from the prepubertal age to adolescence in Spanish children.

AIM: To assess tracking of lipid and apolipoproteins from the prepubertal age (baseline, 6-8 years old) to adolescence (follow-up, 13-16 years old) in Spanish children. METHODS: The sample population included 385 healthy children (179 boys and 206 girls). Tracking was estimated by correlations between baseline and follow-up levels, multiple regression models in which the follow-up lipid was the dependent variable and analysing the percentage of individuals who remained in the same lipid levels status from prepubertal age to adolescence. RESULTS: Correlations between baseline and follow-up levels for low density lipoprotein (LDL)-cholesterol and apolipoprotein B (apo B) were stronger in boys and for high density lipoprotein-cholesterol and apo A-I stronger in girls. Regression analyses showed that, after adjusting by body mass index (BMI), baseline LDL-cholesterol and apo B levels explain 23% and 39% of the variation of follow-up LDL-cholesterol and apo B levels, respectively, in boys and 13% and 22%, respectively, in girls. The strength of tracking for LDL-cholesterol and apo B was 79% and 89%, respectively, in boys and 72% and 82%, respectively, in girls. CONCLUSION: Apolipoprotein B showed the strongest tracking in both sexes, stronger than for LDL-cholesterol, which supports the importance of determining apo B levels as a marker of dyslipidaemia in children.

Litter size affects emotionality in adult male rats.

The role of natural variations in pre-weaning litter size in rodent adult emotionality and the importance of maternal care as a possible mediating factor have been frequently neglected. To address these issues, maternal behaviour of Sprague-Dawley dams differing in natural number of pups was studied for the first seven postnatal days. Later, adult behaviour of representative male offspring was studied in the elevated plus-maze, the circular corridor, the dark-light box and the forced swimming test. Three groups of offspring were selected in function of the number of littermates: L<10 group (less than 10 pups per dam), L10-15 (between 10 and 15 pups per dam) and L>15 group (more than 15 pups per dam). L<10 litters showed a reduced habituation of activity across time in a circular corridor and as compared to L>15 litters, L<10 litters showed a lower activity during the first 5 min of exposure to the circular corridor. L<10 litters had also higher signs of anxiety in the elevated plus-maze, in comparison to the other two groups. In addition, L<10 litters showed in the forced swimming test reduced struggling and more mild swimming behavior than the other two groups. These abnormalities in L<10 litters are not explained by maternal behavior since they received individually more maternal care than L>15, as assessed by total licking-grooming observed during the whole observation period divided by number of pups. Although previous data from several laboratories have demonstrated that low maternal care is associated with heightened emotionality at adulthood, the present results suggest an important contribution of spontaneous litter size to adult emotional behavior that cannot be explained by concomitant changes in maternal care.

Effects of training, early handling, and perinatal flumazenil on shuttle box acquisition in Roman low-avoidance rats: toward overcoming a genetic deficit.

The present series of studies investigated the effects of intensive training, postnatal handling-stimulation and/or perinatal flumazenil (Ro 15-1788, benzodiazepine receptor antagonist) on the acquisition of two-way active avoidance by Roman low-avoidance (RLA/Verh) rats. This rat line has been selectively bred for poor avoidance in the shuttle box, while their Roman high-avoidance counterparts (RHA/Verh) have been selectively bred for their extremely good performance in that task. In the first experiment, RLA/Verh rats submitted to a long and intensive training procedure (unlike those submitted to short training) were able to achieve a performance of 56% of avoidances per session. In the second experiment both postnatal handling and perinatal flumazenil treatments increased avoidance responding in another group of RLA/Verh rats tested at the age of 18 months. Finally, in the last experiment, the performance of a third stock of RLA/Verh rats of the same age which had received perinatal flumazenil did not differ, on the later phases of training, from that shown by RHA/Verh animals. The results are discussed in terms of the "warm up" phenomena which seems to be highly involved in the selection of RLA/Verh rats, as well as on the possibility that central benzodiazepine receptors could play a role in the genetic deficit shown by RLA/Verh rats, which apparently confers a greater emotivity.

Differential effects of early stimulation and/or perinatal flumazenil treatment in young Roman low- and high-avoidance rats.

The effect of infantile handling-stimulation and/or perinatal flumazenil (3.7 mg/kg/day) administration on exploratory and emotional-related behavior was investigated using Roman high- and low-avoidance (RHA/Verh and RLA/Verh) rats. Postnatal handling increased exploration in 30-day-old rats of both psychogenetically selected lines when they were exposed to a hexagonal tunnel maze including an illuminated central arena. Likewise, postnatal stimulation decreased emotional reactivity in both lines of rats, as expressed by increased entry into the central arena, decreased defecation and vocalization frequency, but these effects were more pronounced in the RLA/Verh line. There were interactions between perinatal flumazenil treatment and rat line, indicating that flumazenil enhanced entry into the maze central arena in handled-RLA/Verh rats, whereas a tendency toward the opposite effect was observed in drug-treated and handled-RHA/Verh animals. Thus, the present study emphasizes that the effects of environmental manipulations are partly dependent upon genetic factors, and that pharmacological effects also depend on both genetic and environmentally-induced predisposition.

Infantile stimulation and the role of the benzodiazepine receptor system in adult acquisition of two-way avoidance behavior.

The present study shows that postnatal "consistent" handling (CH) of rats had long-lasting improving effects on coping with an stressful task (i.e. two-way active avoidance), and that such effects were partially prevented by acute Ro 15-1788 (antagonist of benzodiazepine receptor-BZR; 5 mg/kg) administration. Long-lasting detrimental effects in the same task were also observed in rats which received postnatal "inconsistent" handling (INCH), effects that were slightly increased by acute Ro 15-1788 treatment. Finally, Ro 15-1788 tended to increase avoidance acquisition in non-handled (NH) animals. The observed effects of Ro 15-1788 could be partially attributed to a differential modulation of the process of avoidance acquisition depending on postnatal treatments producing different levels of emotionality.

Transmembrane signaling through phospholipase C in cortical and hippocampal membranes of psychogenetically selected rat lines.

RATIONALE: One of the major pathways for neurotransmitter signaling involves phosphoinositide-specific and G-protein-dependent phospholipase C-beta (PLC-beta), which stimulates the formation of inositol 1,4,5-trisphosphate and diacylglycerol. Serotonergic and muscarinic-cholinergic signals in the brain are largely mediated through the hydrolysis of phosphoinositides by PLC. OBJECTIVES: The aim of the experiments reported here was to explore the potential differences in neurotransmitter receptor coupling to PLC in Roman high-avoidance (RHA)/Verh and Roman low-avoidance (RLA)/Verh rats, by examining the changes in agonist (carbachol, 5-methyltryptamine)-stimulated phosphoinositide hydrolysis in hippocampal and cortical membranes derived from the two rat lines. METHODS: To investigate changes in receptor and G-protein coupling to PLC in the brains of these two psychogenetically selected rat lines, which differ in their emotional profiles/learning abilities, we examined GTPgammaS-, agonist (carbachol, 5-methyltryptamine)-, and calcium-stimulated phosphoinositide hydrolysis in cortical and hippocampal membranes of RHA/Verh and RLA/Verh rats. RESULTS: The results indicated that calcium-induced increase in PLC activity was larger in the cortex and hippocampus of RHA/Ver rats, as compared to their RLA/Verh counterparts. Conversely, GTPgammaS- and agonist-induced PLC activity was less pronounced in the hippocampus of RHA/Verh with respect to RLA/Verh rats. Western blot analysis showed no significant differences in the relative values of the G-proteins alphaq/11 and betagamma subunits between both groups of rats in any brain region. However, the levels of PLC-beta1, PLC-beta3, and PLC-beta4 were significantly lower in the hippocampus of RHA/Verh than in RLA/Verh rats. CONCLUSIONS: It is concluded that the hippocampus of RHA/Verh rats has severe deficiencies in PLC activity stimulated by guanine nucleotides and agonists, which are specifically related to a lower level of expression of the PLC-beta type isozymes, a fact that may account for the differential behavioral phenotype observed in these psychogenetically selected rat lines.

Effects of prenatal diazepam on two-way avoidance behavior, swimming navigation and brain levels of benzodiazepine-like molecules in male Roman high- and low-avoidance rats.

Utilizing psychogenetically selected Roman high- and low-avoidance rats (RHA/Verh and RLA/Verh), the present experiments investigated the effects of prenatally administered vehicle and diazepam (1 and 3 mg/kg per day, SC) on the behavior and neurochemistry of adult, male offspring. Active, two-way avoidance behavior was analyzed in 96 rats, at 6 months of age, and swimming navigation in 68 others, at 11 months. Three weeks after testing, selected brain areas from the latter animals were immunoassayed for benzodiazepine (BZD)-like molecules. The 3 mg/kg dose of diazepam both decreased freezing behavior in the shuttle box and reduced the hippocampal content of BZD-like molecules in the RLA/Verh male rats. Swimming navigation (spatial learning), at which the RLA/Verh rats were more adept, was not specifically affected by prenatal diazepam in either rat line. The possibility exists that an increased hippocampal release of BZD-like substances may be necessary to alter shuttle box behavior in RLA/Verh rats.

Environmental enrichment reverses the detrimental action of early inconsistent stimulation and increases the beneficial effects of postnatal handling on shuttlebox learning in adult rats.

Certain types of environmental stimulation administered during critical periods of neural development can enduringly modify adult behavior. The present experiments show that postnatal handling of Sprague-Dawley rats (administered from postnatal days 1 to 22) and/or living in an enriched environment (EE; from weaning until the age of 100 days) clearly improved the ability to learn a two-way active avoidance task in adulthood. In addition the results demonstrated that postnatal inconsistent stimulation (from postnatal days 1 to 22) impaired avoidance acquisition in the same task. This detrimental effect of inconsistent stimulation was reversed by EE. Our findings provide evidence that different types of early experience can influence learning abilities in distinct directions and with different strengths.

Handling-habituation prevents the effects of diazepam and alprazolam on brain serotonin levels in rats.

In two different experiments, serotonin (5-HT) and 5-hydroxyindoleacetic acid (5-HIAA) levels were measured in rats, using HPLC with electrochemical detection, in 3 brain regions (hippocampus, cerebral cortex and hypothalamus) after acute i.p. treatment with diazepam (4 mg/kg), alprazolam (1.25 mg/kg) or vehicle. In the first experiment, rats received the acute treatment 30 min before they were sacrificed. In the second, the animals were previously habituated to handling (involving the maneuvers of injecting and sacrificing at the guillotine) daily for 15 days, before the acute administration of the drugs. Results of the acute treatment alone showed a significant increase in 5-HT levels in hippocampus and cerebral cortex, and a decrease in hypothalamus, but not differences in 5-HIAA levels, for the diazepam- and alprazolam-treated groups. After handling-habituation, no effect in the monoamine or metabolite levels appeared when the rats were treated with diazepam or alprazolam. The results are discussed in relation to the emotional changes induced by the handling procedure, and for possible connections between the mechanisms of action of handling-habituation and benzodiazepine treatments at CNS level.

GABAergic and dopaminergic transmission in the brain of Roman high-avoidance and Roman low-avoidance rats.

The GABAergic and dopaminergic pathways in the central nervous system (CNS) play a pivotal role in the control of emotions and in the adaptive responses to stressful stimuli. The present study was aimed at characterizing a range of biochemical markers of GABA- and dopamine-mediated neurotransmission in the CNS of Roman high-avoidance (RHA/Verh) and Roman low-avoidance (RLA/Verh) rats, two psychogenetically selected lines that differ in their level of emotionality. The stimulatory effect of GABA on 36Cl- uptake was less pronounced in the cerebral cortex of RLA/Verh rats as compared to RHA/Verh rats, whereas no line-related changes were detected in [3H]GABA and [3H]flunitrazepam binding. On the other hand, the density of D1 dopamine receptors labeled with [3H]SCH 23390 was lower in the nucleus accumbens of RLA/Verh rats as compared to their RHA/Verh counterparts, whilst no line-dependent changes were observed in the binding parameters of D1 dopamine receptors in the striatum, amygdala, and prefrontal cortex. These biochemical differences may contribute to the distinct emotionality and responsiveness to the effects of psychoactive drugs of RHA/Verh and RLA/Verh rats.

Long-term behavioural and neuroendocrine changes in Roman high-(RHA/Verh) and low-(RLA-Verh) avoidance rats following neonatal handling.

Roman high-(RHA/Verh) and low-(RLA/Verh) avoidance rats, originally selected and bred for rapid vs poor acquisition of a two-way active avoidance response, differ in emotional reactivity and coping style. These differences are associated with particular neuroendocrine and neurochemical characteristics. New data are presented here to show that the behavioural changes specifically induced by neonatal handling, i.e. decreased emotional reactivity, are associated with marked changes in the neuroendocrine responses of (hyperemotional) RLA/Verh rats to a novel environment. Eight months after neonatal handling, self-grooming behaviour, a reliable marker of emotional reactivity in this line of rats, was significantly decreased in RLA/Verh rats. Defecation scores were also significantly reduced in both lines. Moreover, there was a significant reduction in prolactin and corticosterone release following exposure to a novel environment in neonatally-handled RLA/Verh rats as compared to control, non-handled rats. No effects on prolactin and corticosterone release were observed in RHA/Verh rats. There was also no apparent effect of neonatal handling on coping style i.e. RLA/Verh rats did not increase their spontaneous exploration of novel environments. Thus, the phenotypic expression of basic traits of (high) neuroendocrine/emotional reactivity was specifically modulated by neonatal handling in RLA/Verh rats, whereas both the (hypoemotional) RHA/Verh rats as well as coping style in both lines remained unaffected. Changes in emotional reactivity were still apparent at 12 months of age when rats from the same groups were tested for hyponeophagia. These results suggest that psychogenetically selected lines such as RHA/RLA rats are suitable animal models to investigate interactions between genes and the environment in determining individual sensitivity to stress and coping styles, as well as potential vulnerability (or resistance) to the development of maladaptive syndromes similar to anxiety and mood disorders in humans.

Picrotoxin changes the effects of imipramine and desipramine in rats in the forced swimming test.

The aim was to find whether the chloride channel blocker, picrotoxin, at subconvulsant doses could affect the activity of imipramine or desipramine in the 'forced swimming' test with rats. It was found that picrotoxin enhanced the anti-immobility effects of imipramine and desipramine whereas open field activity remained unaffected or was even decreased by the same treatments. The results seem consistent with recent reports showing direct interactions between several antidepressant drugs and the GABAA receptor/benzodiazepine receptor/chloride ionophore complex (GABAA/benzodiazepine/Cl complex). The results also conform with the hypothesis that a reduction in the functionality of this complex could be related to the clinical effects of antidepressant drugs.

Infantile stimulation and perinatal administration of Ro 15-1788: additive anxiety-reducing effects in rats.

Both postnatal handling of rat pups and perinatal treatment with Ro 15-1788 have been reported to reduce the emotional reactivity of the rats in adulthood, as measured by open field behavior, by the corticosterone response to stress, and by labyrinth and novelty-induced behaviors. We now report results obtained with a well-validated animal model of anxiety (i.e. the elevated plus-maze) that support and extend these previous findings. The results show the clearest reduction of anxiety-related behavior when postnatal handling and perinatal Ro 15-1788 administration were simultaneous.

Sodium valproate reduces immobility in the behavioral 'despair' test in rats.

The present study was conducted to investigate whether sodium valproate could affect immobility in the 'behavioral despair' test in rats. Acute (one injection), subacute (three injections) and chronic treatment with sodium valproate reduced the immobility time in this test, whereas a stimulation of motor activity in the open-field test was not observed with the same drug treatments. The anti-immobility activity of valproate was partially counteracted by the administration of bicuculline (2 mg/kg) or picrotoxin (1.4 mg/kg) before the immobility test. The data agree with previous findings from several animal models of depression of an antidepressant-like activity of GABA mimetics or agents which stimulate GABAergic function.

Pharmacological properties of the GABA(A) receptor complex from brain regions of (hypoemotional) Roman high- and (hyperemotional) low-avoidance rats.

The pharmacological properties of benzodiazepine binding sites of the gamma-aminobutyric acid (GABA)A receptor complex from cortical, hippocampal and cerebellar membranes of Roman high-avoidance (RHA/Verh) and Roman low-avoidance (RLH/Verh) rats were investigated. No major differences between the two lines were found in the binding parameters of [3H]flunitrazepam (a non-selective agonist). [3 H]zolpidem (a Type I selective agonist) or [3 H]ethyl 8-azido-6-dihydro-5-methyl-6-oxo-4H-imidazol[1,5-a]-[1,4]benzodiazepine- 3-carboxylate (Ro15-4513) (a partial inverse agonist). Neither the Kd values nor the Bmax for these ligands differed between RHA/Verh and RLA/Verh rats in any of the brain regions studied. As a result, the proportion of Type I binding sites in cortical and hippocampal membranes of RHA/Verh and RLA/Verh rats or the 'diazepam-sensitive' and the 'diazepam-insensitive' binding sites in cerebellar membranes, calculated from the [3H]flunitrazepam and [3H]zolpidem maximal binding sites or from [3H]Ro15-4513 binding (in the absence or in presence of diazepam), respectively, was also similar. Furthermore, there were no differences between the two rat lines in the allosteric interactions between GABA and the benzodiazepine binding sites (labeled with [3H]flunitrazepam) in all three areas tested or the Type I binding sites (labeled with [3H]zolpidem) in the hippocampus. In contrast, RLA/Verh rats showed a significant reduction in the allosteric interactions between GABA and [3H]zolpidem binding sites in the cortex. As a whole, these results indicate the absence of generalized between-line differences in the GABA(A) receptor complex showing, at the same time, the existence of some specific differences in allosterism within the GABA(A) complex. These differences may contribute to the divergent emotional responses which characterize the RHA/Verh and RLA/Verh rat lines.

Early stimulation effects on novelty-induced behavior in two psychogenetically-selected rat lines with divergent emotionality profiles.

The present study shows that postnatal handling (H: consisting of removing the pups from the nest twice daily and placing them individually in plastic cages lined with paper towel for a period of 10 min, between postnatal days 1 and 21) and/or environmental enrichment (E: for a period of 6 months) of Roman high- and low-avoidance (RHA/Verh and RLA/Verh) rats induced long-lasting decreases in emotional reactivity (i.e. reduced defecations in the open field, OF, and hole-board, HB, tests) as well as increases in exploratory behavior (i.e. head-dipping) in a manner dependent upon the rat line (there were 'line x H' and 'line x E' interactions). It is reported for the first time that RHA/Verh rats show more head-dipping behavior than RLA/Verh rats, and that the environmental treatments can increase head-dipping of RLA/Verh animals to the level shown by RHA/Verh rats.

Beneficial effects of infantile stimulation on coping (avoidance) behavior in rats are prevented by perinatal blockade of benzodiazepine receptors with Ro 15-1788.

The present study shows that postnatal 'consistent' handling (CH; which consisted of removing the pups from the nest and placing them individually in plastic cages lined with paper towel for a period of 15 min daily between 1 and 22 postnatal days) of rats had long-lasting improving effects on coping with a stressful task (i.e. enhancement on the early acquisition of two-way active avoidance), but such effects were completely prevented when CH treatment was combined with chronic perinatal Ro 15-1788 (7 mg/kg/day, between prenatal day 19 and postnatal day 22) administration (i.e. blockade of benzodiazepine receptor (BZR)). A long-lasting decremental effect was also observed in the same task in rats which received postnatal 'inconsistent' handling (INCH; in which stimulation of pups was changed every day between 1 and 22 postnatal days), without being affected by the concomitant perinatal Ro 15-1788 treatment. These results suggest that intact ontogeny of BZRs is necessary to obtain the enduring positive effects of CH on emotional behavior (i.e. early acquisition of two-way active avoidance).

Impaired short- and long-term memory in Ts65Dn mice, a model for Down syndrome.

Ts65Dn (TS), control littermates (CO) and Swiss (SW) male mice were tested in the elevated plus-maze and in the Morris water maze (MWM) for memory evaluation. In the plus-maze, each mouse was placed at the end of an open arm and initial freezing and the time to enter into an enclosed arm (transfer latency) were measured. SW mice decreased both measures over repeated trials, whereas no decrease of freezing was observed in CO mice, thus suggesting increased emotionality in this group. Compared to CO mice, TS mice showed less initial freezing, shorter transfer latencies, and spent less time in enclosed arms, suggesting a reduced ability to habituate or to inhibit behaviour. Animals were also submitted to a learning-set paradigm consisting of reaching a new platform position each day in the MWM. Two training phases (separated by a resting period of 6 weeks), each including eight acquisition and four cued sessions, were performed (each session consisting of four pairs of trials). CO and SW mice already reached an asymptotic performance by the second day of the first phase whereas TS mice did not achieve that level until the second training phase. The progression over trials indicated that CO and SW animals learned the new platform position between trials 1 and 2 of each session, whereas TS animals failed to do it and had more difficulties to find the platform when it was placed in the centre of the pool as compared to the other positions (SW, NE, E). The results suggest that TS mice show working memory impairments in addition to long-term memory deficits, although extensive training appeared to facilitate TS mice to achieve a level of performance similar to their control littermates. This represents another aspect of the cognitive deficits shown by TS mice: a mouse model of the human Down syndrome.

A behavioral assessment of Ts65Dn mice: a putative Down syndrome model.

Mice which are trisomic for only the human chromosome (Chr) 21-homologous segment of mouse Chr 16 (segmental trisomy), including a portion of the Down syndrome region of human Chr 21, have recently been developed. Since these segmentally trisomic mice, designated Ts(17(16))65Dn, survive to adulthood, they may represent a mouse model for the study of Down syndrome. A partial characterization of their behavioral phenotype was undertaken by evaluating the sensorimotor reflexes, exploration, locomotor activity, emotionality and spatial learning in 16 male Ts65Dn mice (TS) and 16 control (CO) littermates. No sensorimotor deficits appeared in TS compared to CO mice. By contrast, head-dipping behaviour in the hold board was increased in TS mice with respect to the CO group, showing a higher repetition rate of previously explored holes. Crossings in the open field and total arm entries in the plus maze were higher in TS than in the CO group during the dark phase of the light-dark (LD) cycle under red light, but not during the light phase of the LD cycle under white light. Entries into the open arms of the plus maze were increased overall in TS mice when compared to CO mice, but no differences were found in time spent in the open arms. TS mice showed impaired place learning in the Morris water maze, whereas they were able to reach the same performance as CO animals in cued learning. Thus, absence of sensorimotor deficits, increased exploration, hyperactivity under certain experimental conditions and a moderate impairment of spatial learning were the principal characteristics observed in TS mice compared to their CO littermates.