The effects of environmental enrichment and social isolation and their reversion on anxiety and fear conditioning.

Animal models of fear and anxiety provide important insight into anxiety-related symptoms in humans. Environmental physical conditions and social contact influence behavior and brain plasticity particularly at early developmental stages and have long lasting effects reaching even adulthood. The potential benefit that a later environmental enrichment may have on rats raised in isolation is however not fully understood. We aim to investigate the effects of housing conditions and their reversion on anxiety and fear-related behaviors in rats. In phase I, we compared the effects of different housing conditions (environmental enrichment, control and social isolation) on anxiety behaviors in the open field test, elevated plus maze and fear conditioning. We found stronger effects of housing on behavioral tests when induced at weaning (phase I), than later in development (phase II). After one month, EE rats showed lower anxiety related behaviors and more freezing in FC. In phase II, we evaluated the effects of the reversion of housing conditions on the same behavioral parameters. We observed a behavioral trend such that the groups started to behave similar to their new housing conditions in OFT mainly. These results suggest that housing conditions at weaning can have long-lasting effects on anxiety and fear-related behaviors. Because the behavioral changes observed after the housing reversal in adulthood were partial, we suggest that more time of social and physical enrichment could be necessary to promote major changes in behavior at this age.

Age-dependent effects of environmental enrichment on spatial memory and neurochemistry.

Although aging and environmental stimulation are well-known to affect cognitive abilities, the question of whether aging effects can be distinguished in already-mature adult rats has not been fully addressed. In the present study, therefore, young and mature adult rats were housed in either enriched or standard conditions (EE or SC) for three months. Open-field (OFT) and radial-maze (RM) behavior, and ex-vivo contents of GABA and glutamate in hippocampus, and of dopamine and DOPAC in ventral striatum (VS) were analyzed and compared between the four groups. In OFT, young rats were more active than mature adults irrespective of the housing condition. Surprisingly, in the RM test, mature adults outperformed young counterparts except for the young-enriched rats, which showed a progressive improvement in RM performance. At the neurochemical level, young EE rats showed higher hippocampal glutamate and GABA concentrations, and DA turnover in VS, which correlated with RM performance. Altogether, the behavioral and cognitive strategies underlying habituation learning and spatial memory seem to be qualitatively different between the two ages analyzed. These results challenge the assumption that mature adult animals are always worse in learning and memory tasks. However, young rats benefited more from the social and physical stimulation provided by the enrichment than mature adult counterparts. The latter effect was evident not just on behavior, but also on brain neurochemistry.

Individual differences in the forced swimming test and neurochemical kinetics in the rat brain.

Individual differences in the forced swimming test (FST) could be associated with differential temporal dynamics of gene expression and neurotransmitter activity. We tested juvenile male rats in the FST and classified the animals into those with low and high immobility according to the amount of immobility time recorded in FST. These groups and a control group which did not undergo the FST were sacrificed either 1, 6 or 24 h after the test. We analyzed the expression of the CRF, CRFR1, BDNF and TrkB in the prefrontal cortex, hippocampus and nucleus accumbens as well as norepinephrine, dopamine, serotonin, glutamate, GABA and glutamine in the hippocampus and nucleus accumbens. Animals with low immobility showed significant reductions of BDNF expression across time points in both the prefrontal cortex and the nucleus accumbens when compared with non-swim control. Moreover, rats with high immobility only showed a significant decrease of BDNF expression in the prefrontal cortex 6h after the FST. Regarding neurotransmitters, only accumbal dopamine turnover and hippocampal glutamate content showed an effect of individual differences (i.e. animals with low and high immobility), whereas nearly all parameters showed significant differences across time points. Correlational analyses suggest that immobility in the FST, probably reflecting despair, is related to prefrontal cortical BDNF and to the kinetics observed in several other neurochemical parameters. Taken together, our results suggest that individual differences observed in depression-like behavior can be associated not only with changes in the concentrations of key neurochemical factors but also with differential time courses of such factors.

Individual differences in the immobility behavior in juvenile and adult rats are associated with monoaminergic neurotransmission and with the expression of corticotropin-releasing factor receptor 1 in the nucleus accumbens.

The study of individual differences provides an important methodological approach to analyze the neurobehavioral spectrum of a given cohort in order to understand brain function and disease. Based on immobility time in the forced swimming test (FST) juvenile and adult rats were classified as subgroups with low and high immobility. Afterwards, we compared behavior, neurochemical parameters, and gene expression profiles in some brain areas of rats with low and high immobility only. No differences in the open field test (OFT) were observed between subgroups. Regarding neurochemistry, juvenile animals with low immobility showed higher accumbal dopamine turnover and lower hippocampal norepinephrine concentrations, whereas adult rats only differed for accumbal dopamine, although in an opposite direction from that observed in juveniles. Moreover, the expression of accumbal corticotrophin-releasing factor receptor 1 (CRFR1) was significantly different in animals with low and high immobility at both ages, with animals less immobile showing higher levels of CRFR1 mRNA levels. Taken together, our findings suggest that differences in monoaminergic neurotransmission and CRFR1 expression are associated with the coping strategy adopted by the animal and with the tendency to develop depression-related behaviors. Concerning monoaminergic neurotransmission such association is modulated by age, and such modulation could be related to the differential behavioral results observed between juvenile and adult rats.

Behavioral and neurochemical characterization of maternal care effects on juvenile Sprague-Dawley rats.

Maternal care represents a major constituent of early life environment and has the potential to modulate critical neurobehavioral responses to stress. The aim of the present study was to determine the effects of naturally occurring variations in maternal care on behavioral and neurochemical responses of juvenile Sprague-Dawley rats. A group of dams were classified based on their licking behavior in high and low licking-grooming mothers. Afterwards, the male offspring was tested in a series of behavioral tests: open field test (OFT), elevated plus maze (EPM) and forced swimming test (FST). Additionally, monoamine concentrations were determined post-mortem in three brain regions: hippocampus, ventral striatum and prefrontal cortex. Our findings suggest that maternal care variations have an effect on several anxiety-related behaviors in OFT and EPM but not in depression-like behaviors in FST. Such behavioral differences could be related to an increased DOPAC concentration and 5-HT turnover in prefrontal cortex. These evidences suggest that natural variations in maternal care modified some behavioral and neurochemical parameters related with anxiety and stress in this strain.

Maternal behavior as an early modulator of neurobehavioral offspring responses by Sprague-Dawley rats.

Maternal care plays an important role as an early modeler of neurodevelopment and brain function, and its effects remain until adulthood. Such modeling or programming has shown to influence the stress response and represents a key susceptibility factor in the development of mood disorders. In order to characterize such process which is still not clear, male offspring were classified in animals with low, medium and high licking/grooming (LG) according to the maternal behavior. Juvenile animals were subjected to the open field test (OFT) and the forced swimming test (FST), and offspring of low and high LG mothers were compared. Seven days after the FST, neurochemical and gene expression analyses were carried out in order to identify possible changes on relevant targets. Maternal care did determine locomotor behaviors in the OFT, supporting an anxiogenic effect of low maternal investment. This effect seems to be associated with the serotonergic systems in both nucleus accumbens (NAc) and hippocampus (HPC), since offspring of low LG mothers showed decreased 5-HT neurotransmission in those brain regions compared with animals of high LG mothers. Furthermore, TrkB expression was higher in offspring of high LG compared to the group of low LG mothers, supporting its influence as a mechanistic intermediate of such effect, at least in the NAc. Taken together, these findings strongly support the influence of differential maternal care on the neurodevelopment and responsivity of juvenile rats.