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Effect of prenatal ultrasound exposure on adult behavior in mice.

Pregnant Swiss mice were exposed to diagnostic levels of ultrasound (3.5 MHz, Maximum acoustic output: ISPTP = 1 W/cm2 and ISATA = 240 mW/cm2, acoustic power = 65 mW) for 10 min on days 11.5 or 14.5 postcoitus (PC). At 3 and 6 months postpartum, offspring were subjected to the following behavioral tests: bright and dark arena test for locomotor/exploratory activity and passive avoidance test for learning and memory. Anxiolytic activity and latency in learning were noticed in the ultrasound-treated animals. The effect was more pronounced in the 14.5 days PC group than in the 11.5 days PC group. But memory was not affected in the ultrasound-exposed animals. There was a nonsignificant decrease in the total locomotor activity at 6 months of age in all the exposed animals. Thus, the present data demonstrate that exposure to diagnostic ultrasound during late organogenesis period or early fetal period in mice may cause changes in postnatal behavior as evidence by selected adult offspring behavioral tests. However, any conclusive statement in this regard should await results from more detailed investigations.

Animals↗

Home cage behavior and lead treatment in rhesus monkeys: a comparison with open-field behavior.

Nursery-reared rhesus monkeys were treated with no (n = 4) or moderate levels of lead (n = 4) during the first postnatal year. Mean blood lead levels peaked at 55 micrograms/dl at 5 weeks of age, averaged 36 micrograms/dl for the remainder of the first year postpartum, and declined to < or = 5 micrograms/dl for the 4 monkeys by 2.3 years of age. Previously, the lead-treated monkeys exhibited increased environmental exploration and decreased inactivity in a nonhuman primate version of the open field when tested at 4, 5, and 6 years of age (5,6). The current study was designed to assess behavior in the home cage of these monkeys at 6 years of age to determine: (a) whether the increased exploration was specific to the open field, and (b) any lead-related behavioral alterations specific to the home cage. Each monkey was observed twice weekly for 10 weeks and the duration and frequency of 17 behaviors were recorded. Lead treatment did not result in significant alterations in any of the six behaviors which occurred with enough frequency to warrant analysis. As a whole, all monkeys were either inactive or engaged in self-grooming for a large proportion of the test session. Less frequent were behaviors such as locomotion, environmental exploration, and self-directed behaviors. The distribution of behavioral activities in the home cage differed from that in the open field. Potential reasons for the expression of significant lead-related effects in the open field and not in the home cage are discussed as well as the differences in distribution of behavioral activities.

Animals↗

Medial prefrontal cortex is involved in spatial temporal order memory but not spatial recognition memory in tests relying on spontaneous exploration in rats.

The present study describes two novel tasks relying on spontaneous patterns of exploration in a radial-arm maze that can be used to assess spatial recognition memory and spatial temporal order memory (i.e. memory for the order in which places have been visited) in the rat. In the recognition memory task, rats were permitted to freely explore two arms in the maze on a first trial and one 'familiar' arm and one novelly located arm on a second trial 105 min later. In the temporal order memory task, rats were permitted to explore two arms in the maze on a first trial, two novel arms on a second trial 60 min later, and one 'older familiar' arm and one 'more recent familiar' arm on a third trial 45 min later. Using these tasks, we found that rats direct greater exploration at a novel than a familiar arm location, thus showing long-term spatial recognition memory, and at an older familiar arm than a more recent familiar arm, thus showing long-term spatial temporal order memory. Lidocaine inactivation of the mPFC prior to the final trial in each task disrupted performance on the temporal order but not the recognition memory task, thereby demonstrating a role for the mPFC in the retrieval and/or use of temporal order information but not in spatial memory per se. These findings highlight the specific involvement of the rat mPFC in temporal order memory and have important implications for a broader understanding of mPFC function.

Analysis of Variance↗

Multidimensional structure of anxiety-related behavior in early-weaned rats.

Early environmental stimuli affect various aspects of physical and behavioral development. Weaning is one of the most important events in the early stage of life, and recently we have found that precocious weaning augments anxiety and aggressiveness in mice. Here, we report the presence of virtually identical phenomena in rats. To understand the multidimensional structure of anxiety-related behavior, the influence of early weaning upon behavior in adulthood was investigated using three behavioral tests: the elevated plus-maze test, the hole-board test and the open-field test. Two groups of rats were prepared. One was weaned from the dam at 16 days of age (early-weaned group) and the other at 30 days (normally weaned group) as a control. Both groups were subjected to the three tests at 8-10 weeks of age. The elevated plus-maze test revealed lower frequency of entry to and shorter duration of stay in the open arms in the early-weaned animals. In the hole-board test, the early-weaned rats showed lower frequency and shorter duration of head dipping into the holes. And in the open-field test, the early-weaned rats tended to stay at the central square for a shorter period and to defecate more frequently. The behavioral parameters of the three tests were combined and subjected to principal component analysis (PCA). The factorial scores for six extracted factors were compared between the early-weaned and normally weaned groups, and it was revealed that the early-weaned rats had a lower score in Factor 1 (non-anxious exploration) and Factor 5 (risk assessment behavior). Taken together, these results suggest that the time of weaning had a considerable impact on behavioral development, particularly with respect to anxiety-related behaviors.

Age Factors↗

Early social and physical deprivation leads to reduced social motivation in adulthood in Wistar rats.

Behavioural abnormalities in adulthood may have their origin in a disturbed interaction with the environment during postnatal development. We tested the consequences for adult social motivation of early deprivation (ED) of rat pups from mothers and littermates relative to nonhandled (NH) pups. Early deprivation was performed at room or warm ambient temperatures, cold-ED and warm-ED, respectively, and during either the dark or light phase of the daily cycle. In adulthood, rats that were unrelated and unfamiliar but of the same treatment group were introduced in pairs to an open field for a 30-min test. Social behaviour in home base and exploration modes was assessed using algorithmic analysis of the XY locations of the two rats. Findings revealed that Cold-ED induced a preference for a separate home base, which limited significantly the episodes of social interactions, in comparison to NH. Warm-ED had no comparable effect on the rats' social behaviour. These findings indicate that ED under ambient conditions that constitute severe thermal stress for rat pups leads to development of reduced social motivation in adulthood.

Analysis of Variance↗

Contrasting grooming phenotypes in three mouse strains markedly different in anxiety and activity (129S1, BALB/c and NMRI).

129S1/SvImJ (129S1), NMRI and BALB/c mice are widely used in behavioural research, demonstrating marked strain differences in their behavioural phenotypes. Grooming is a complex and essential ritual in the rodent behavioural repertoire with a general cephalocaudal progression (forepaws-nose-face-body-legs-tail and genitals). Various stressors as well as genetic manipulations have been reported to alter mouse grooming and its patterning, underlying the importance of analysis of grooming behaviours. Although strain differences between these mice have been assessed in many studies, no comparative analyses of their grooming have been performed. Here we show strain differences in spontaneous (novelty-induced) grooming between 129S1, NMRI and BALB/c mice. Overall, 129S1 mice demonstrated lower grooming activity and impaired microstructure (more interrupted bouts and incorrect transitions contrary to the cephalocaudal rule), accompanied by lower vertical exploration. In contrast, BALB/c and NMRI mice showed high vertical activity and unimpaired grooming microstructure, also exhibiting different grooming levels (BALB/c>NMRI). Our study suggests that contrasting grooming phenotypes in these mice may not be due to the strain differences in their sensory abilities, general activity levels, brain anatomy or aggressiveness, but rather reflect a complex interplay between anxiety, motor and displacement activity in these strains (hypoactive anxious phenotype in 129S1 mice, active anxious phenotype in BALB/c and non-anxious high displacement phenotype in NMRI mice). We suggest that ethological analysis of mouse grooming, such as that reported here, may be a useful tool in neurobehavioural research.

Analysis of Variance↗

Rat strain differences in sleep after acute mild stressors and short-term sleep loss.

Genetic and physiological diversity amongst rodent strains provide the potential for developing models that may give insight into factors that regulate sleep in response to environmental challenges. We examined home cage activity, behavioral performance in the open field and sleep after a number of mild stressors (cage change [CC], open field [OF]) and after 1 and 4h of sleep deprivation (1hSD and 4hSD) in rat strains (Fischer 344 [F344], Lewis [LEW], Wistar [WST] and Sprague-Dawley [Sp-D], n=16 per strain) that differ in behavior and sleep. F344 and WST rats had greater home cage locomotion than LEW and Sp-D rats, but F344 rats exhibited the least relative locomotion in OF. In 24h baseline recordings of sleep, strain rankings were LEW=WST=Sp-D>F344 in rapid eye movement sleep (REM), and LEW=Sp-D>F344 and LEW>WST in non-REM (NREM). Compared to baseline, total sleep was reduced in all four strains after CC, OF and 1hSD, but not after 4hSD, in the first hour after treatment. Afterwards, increases in REM and NREM were seen after all treatments with the amount and time course varying across treatments and strains. CC induced the weakest and 4hSD the largest effects on sleep, whereas OF and 1hSD had intermediate effects. Among strains, the more anxious F344 rats exhibited the greatest sleep increases during the light period after OF, 1hSD and 4hSD. The results are discussed with respect to the relationship between behavioral and sleep responses to stressors, and to potential mechanisms underlying the strain differences.

Analysis of Variance↗

Prenatal stress produces more behavioral alterations than maternal separation in the elevated plus-maze and in the elevated T-maze.

Prenatal stress and maternal separation are used in a large number of studies on early adversity consequences and present some similarities in their effects. The present work investigates the behavioral effects of these two procedures on two models of anxiety: the elevated plus-maze and the elevated T-maze. During pregnancy, female rats were submitted to uncontrollable electric foot shock sessions every other day or kept undisturbed. After delivery, litters from undisturbed dams were submitted to either 180-min daily periods of maternal separations from the 3-14th postnatal days or maintained with the dams all the time. Litters from the stressed dams were left undisturbed from the 3-14th postnatal days. Only males were tested. In adulthood, rats were tested in the elevated T-maze or in the elevated plus-maze. In the latter procedure half the subjects were submitted to a 60-min period of restraint immediately before being tested. The following measures were taken in the elevated plus-maze: frequency and time spent in entries into the arms, stretching, rearing, grooming and head dipping. In the T-maze measures of avoidance and escape latencies were used. Our data indicated that prenatal stress had more pronounced anxiogenic effects than maternal separation, as judged by reduced exploration of the open arms of the elevated plus-maze, but mainly after the restraint stress, and increase in avoidance latencies in the elevated T-maze. The other measures not directly involved in the elevated plus-maze arm exploration yielded similar results. Our data indicate that prenatal stress causes more anxiogenic effects in adulthood than maternal separation but, in the elevated plus-maze, these anxiogenic effects are better seen immediately after an acute stress.

Animals↗

Pharmacological treatment to augment hole board habituation in prenatal Vitamin D-deficient rats.

Neurocognitive impairment has consistently been considered a central and stable feature of schizophrenia. There is much controversy about the effects of neuroleptics on neurocognitive deficits. Thus, further investigations are needed to clarify the pathological substrate of cognitive deficits in schizophrenia as well as to identify pharmacological tools for treatment. Transient prenatal Vitamin D deficiency is considered a developmental model in schizophrenia research. Recently, it was reported that prenatal Vitamin D-depleted rats showed a habituation deficit in the hole board. Here, we tested the effect on hole board habituation of haloperidol (Hal, 0.075 mg/kg, i.p.), risperidone (Ris, 0.2 mg/kg, i.p.) and the mGluR5 agonist CHPG (0.1 mg, i.c.v.) after subchronic treatment. Hal was found to impair habituation in control animals, Ris restored hole board habituation, whereas Hal and CHPG normalised hole board habituation in the deplete animals completely. The results of the study demonstrate that (i) the Vitamin D model might be a valuable tool in the study of neurodevelopmental aspects of schizophrenia, (ii) the model is sensitive in detecting the effect of antipsychotic drugs and (iii) the model appears to be sensitive in differentiating between typical and atypical antipsychotic drug.

Animals↗

Heterozygous mice with Ric-8 mutation exhibit impaired spatial memory and decreased anxiety.

Ric-8 is a guanine nucleotide exchange factor for a subset of Galpha proteins and it is required to maintain Galpha(q) and the Galpha(s) pathways in functional state. In adult mice Ric-8 is expressed in regions involved in the regulation of behavior (neocortex, cingulate cortex and hippocampus). As Ric-8 is shown to regulate neuronal transmitter release, the aim of present study was to perform behavioral analysis of ric-8 mutant. Homozygous (-/-) ric-8 mutant mice are not viable and die in early embryonic development, therefore for behavioral analysis heterozygous (+/-) ric-8 mutant mice were used. We found decreased anxiety of ric-8 heterozygous mice in light-dark compartment test where mutant mice significantly avoided the light compartment. In spatial learning paradigm (Morris water maze) the performance of ric-8 (+/-) mice was impaired. Namely, in the reversal test, ric-8 (+/-) mice exhibited significant delay to find the hidden platform compared to wild-type (wt) littermates. We did not find differences in the behavioral tests reflecting the motor abilities of mice (motor activity, rota-rod). Therefore, described alterations seem to be specific for anxiety and spatial learning. Based on these results we can conclude the importance of ric-8 in the regulation of memory and emotional behavior.

Animals↗

Individual differences in novelty-seeking behavior but not in anxiety response to a new environment can predict nicotine consumption in adolescent C57BL/6 mice.

Considering that adolescence is associated with an increased motivation to seek out new stimuli and with low anxiety levels in exploring novel environments, and that both behavioral traits may be associated with substance abuse, we investigated whether the behavioral response to a novel environment can predict subsequent oral nicotine self-administration in adolescent C57BL/6 mice. On postnatal day 30 (PN30), the novelty-seeking behavior and anxiety levels were assessed in a hole board activity box. The total number of head-dips (DIP) was used to classify animals either into the high novelty (HN; DIP above median) or low novelty (LN; DIP below median) groups. The percentage of center squares crossed (CEN) was used to classify animals either into the high anxiety (HA; CEN below median) or low anxiety (LA; CEN above median) groups. From PN31 to PN41, all animals were given a free choice between tap water or a nicotine solution (10 microg/ml). LN mice did not change nicotine intake throughout the free choice procedure, however, HN mice presented a marked increase in consumption. There were no differences in consumption between HA and LA mice. Our results indicated that mice that presented a more intense novelty-seeking behavior increased their preference for nicotine during the free choice experiment but that anxiety levels did not predict nicotine consumption. These results suggest that higher motivation to seek out new experiences is a significant contributor to drug use in adolescents and that anxiety is probably not a major factor that determines differential nicotine consumption during adolescence.

Age Factors↗

Comparisons between anxiety tests for selection of anxious and non anxious mice.

Male Swiss albinos mice were submitted to two behavioural tests intended to determine their anxiety level: the elevated plus-maze test as well as the black and white compartments test. In addition they were submitted to the hole-board test. It was observed: (i) that the correlation between scores in the two first tests was weak, suggesting that they explore different components of anxiety; (ii) that the score on the latter test was better correlated with the response in the elevated plus-maze test than in the black and white compartments test. From these data three groups of animals were constituted, considered, respectively, as anxious, non anxious and intermediates. It was observed that both horizontal and vertical locomotion in an unfamiliar environment differed between groups, with higher activity in non anxious than in anxious. In the hole-board test, only animals classified as anxious displayed an obvious response to the anxiolytic drug diazepam (0.5mg/kg). Finally in the forced-swimming test, the three groups demonstrated a similar immobility time, suggesting that the operated segregation was not depending on a helpless component. It is proposed that the selection of mice from a combination of either elevated plus-maze and black and white compartments tests or a combination of hole-board test and black and white compartments test, allows to distinguish high or low anxiety animals among a population of mice.

Analysis of Variance↗

Twice daily long maternal separations in Wistar rats decreases anxiety-like behaviour in females but does not affect males.

Prolonged daily separations of rat pups from their mother have been reported to increase anxiety-like behaviour in adult offspring. However, there are an increasing number of studies not showing this. It has been proposed that the effect of long maternal separation (LMS) is partly due to the disruption of maternal care caused by the separations. The aim of the present study was to investigate whether increasing the number of daily separations would produce more robust effects in the adult offspring on anxiety-like behaviour in the defensive withdrawal test, and on spontaneous motor activity. Since previous studies of LMS have revealed sex differences in behaviour, we included both males and females. In our separation paradigm we subjected rat pups to either two daily 3h maternal separations during the first 2 weeks postpartum (LMS), two daily 15 min maternal separations (brief maternal separations, BMS) during the same time period to control for the effects of handling, or to normal husbandry conditions. As adults we found no effects of this LMS paradigm in male rats, although BMS males showed a tendency toward decreased anxiety-like behaviour. In contrast, LMS females showed a decrease in anxiety-like behaviour. We also found significant sex differences that were most prominent in the LMS group, indicating that females are more sensitive to our maternal separation paradigm. The present study suggests that increasing the number of maternal separations does not increase anxiety-like behaviour in neither male nor female Wistar rats.

Animals↗

Hyperlocomotion associated with transient prenatal vitamin D deficiency is ameliorated by acute restraint.

Transient prenatal vitamin D deficiency produces hyperlocomotion in the adult rat. The aim of this study was to examine the effects of acute restraint on the behaviour of DVD and control rats in the open field. Rats were conceived and born to developmentally vitamin D (DVD) deficient or replete (control) dams and, at 8 weeks of age, were monitored for 30 min in an open field using automated video tracking software. Half of the rats were restrained within a towel for 5 min immediately before the open field test. The remainder received minimal handling prior to the open field test. Repeating previous findings, DVD deficient animals had enhanced locomotion during the first 10 min of the open field test compared to control rats. By contrast, there were no differences in locomotor activity after acute restraint stress. The time rats spent in the corners and side of the open field was affected by prenatal diet. DVD rats spent less time in the corners and more time in the side than control rats across the whole 30 min test. This difference was not seen in rats with acute restraint stress. The time spent in the centre was not altered by prenatal diet or acute restraint. Thus, transient prenatal vitamin D deficiency induces a transient spontaneous hyperlocomotion in adulthood that is modulated by acute restraint stress.

Analysis of Variance↗

Over-expression of two different forms of the alpha-secretase ADAM10 affects learning and memory in mice.

Members of the ADAM family (adisintegrin and metalloprotease) are the main candidates for physiologically relevant alpha-secretases. The alpha-secretase cleaves in the non-amyloidogenic pathway the amyloid precursor protein within the region of the Abeta peptides preventing their aggregation in the brain. The increase of alpha-secretase activity in the brain provides a plausible strategy to prevent Abeta formation. Concerning this possibility two transgenic mouse lines (FVB/N) have been created: mice over-expressing the bovine form of the alpha-secretase (ADAM10) and mice over-expressing an inactive form of the alpha-secretase (ADAM10-E348A-HA; ADAM10-dn). For behavioral examination a F1 generation of transgenic mice (C57Bl/6 x FVB/N (tg)) was generated and compared to wild type F1 generation (C57Bl/6 x FVB/N). Behavior was characterized in the following tasks: standard open field, enriched open field, elevated plus-maze, and the Morris water maze hidden platform task. Concerning basal activity, exploration, and anxiety, transgenic mice behaved similar to controls. With respect to learning and memory both transgenic lines showed a significant deficit compared to controls. ADAM10 mice however, showed thigmotaxis with passive floating behavior in the Morris water maze indicating differences in motivation, whereas, ADAM10-dn mice displayed an inconspicuous but limited goal-directed search pattern. Thus variation of the enzymatic activity of alpha-secretase ADAM10 alters learning and memory differentially. Nevertheless, it could be concluded that both, ADAM10 and ADAM10-dn mice are suitable control mice for the assessment of alpha-secretase-related effects in animal models of Alzheimer's disease.

ADAM Proteins↗

Behavioral inhibition and impaired spatial learning and memory in hypothyroid mice lacking thyroid hormone receptor alpha.

Thyroid hormone insufficiency leads to impaired neurogenesis, behavioral alterations and cognitive deficits. Thyroid hormone receptors, expressed in brain regions involved in these behaviors, mediate the effects of thyroid hormone deficiency or excess. To determine the contribution of thyroid hormone receptor alpha (TRalpha) in these behaviors, we examined the behavior of euthyroid as well as hypo- and hyperthyroid mice lacking all isoforms of the TRalpha (TRalpha(o/o)). The hypothyroxinemic TRalpha(o/o) mice demonstrated behavioral inhibition, manifested in decreased activity and increased anxiety/fear in the open field test (OFT) and increased immobility in the forced swim test (FST) compared to C57BL/6J mice. TRalpha(o/o) mice also showed learning and recall impairments in the Morris water maze (MWM), which were exaggerated by hypothyroidism in TRalpha(o/o) mice. These impairments were concurrent with increased thigmotaxis, suggesting an increased anxiety-like state of the TRalpha(o/o) mice in the MWM. Expression of genes, known to be involved in processes modulating learning and memory, such as glucocorticoid receptor (GR), growth-associated protein 43 (GAP-43) and neurogranin (RC3), were significantly decreased in the hippocampus of TRalpha(o/o) mice. GR expression was also decreased in the frontal cortex and amygdala of TRalpha(o/o) mice, indicating that expression of GR is regulated, probably developmentally, by one or more isoforms of TRalpha in the mouse brain. Taken together these data demonstrate behavioral alterations in the TRalpha(o/o) mice, indicating the functional role of TRalpha, and a delicate interaction between TRalpha and TRbeta-regulated genes in these behaviors. Thyroid hormone-regulated genes potentially responsible for the learning deficit found in TRalpha(o/o) mice include GR, RC3 and GAP-43.

Analysis of Variance↗

Regulatory mechanisms underlying novelty-induced grooming in the laboratory rat.

Bouts of pelage cleaning can be readily evoked in laboratory rodents under conditions of exposure to novelty. Such novelty-induced grooming is described as stereotyped and rostro-caudal in its progression. The patterned structure of novelty-induced grooming makes it particularly attractive for research on the organizational and motivational underpinnings of co-ordinated behaviour. Micro-characteristics of stereotyped novelty-induced grooming bouts were studied in 27 female Wistar rats that were exposed to a novel arena with shelters for a period of 15 min. The order of grooming acts within the initial bouts was rostro-caudal, but subsequent bouts became progressively disorganized in their sequencing. The observed pattern of progressive bout disorganization may be attributed to the gradual dearousal from stress. Differences between consecutive bouts in their micro-characteristics suggest that at least some grooming actions emitted within the context of those bouts operate as relatively independent units of behaviour. Those unitary component actions appear to be integrated into protracted pelage cleaning sequences by a separate mechanism. Similar endogenous mechanisms have been proposed for other co-ordinated motor actions, which suggests that the organizational principles identified in the context of novelty-induced grooming may represent general principles that govern co-ordinated behaviour.

Animals↗

Impaired motor performance in mice lacking neurosteroid vitamin D receptors.

Vitamin D is a neuroactive seco-steroid and its importance to the nervous system is receiving increasing recognition. Since numerous data link vitamin D dysfunctions to various neurological and behavioural disorders, we studied whether genetic ablation of vitamin D receptors (VDR) may be associated with motor impairments in mice subjected to several behavioural tests. The data obtained in the vertical screen and swim tests show that VDR genetic ablation produces severe motor impairment (shorter screen retention and poor swimming) in mutant mice compared to wild-type and heterozygous control animals. These impairments appear to be unrelated to visual, vestibular and activity/emotionality parameters of mice, and are likely associated with disturbed calcium homeostasis. This study confirms the important role of the vitamin D system in motor functions and suggests that animal genetic models targeting the vitamin D/VDR system may be a useful tool to study vitamin D-related motor/behavioural disorders.

Animals↗