Environmental Enrichment Protocol Research Articles

Microglial Rac1 is essential for experience-dependent brain plasticity and cognitive performance

Microglia, the largest population of brain immune cells, continuously interact with synapses to maintain brain homeostasis. In this study, we use conditional cell-specific gene targeting in mice with multi-omics approaches and demonstrate that the RhoGTPase Rac1 is an essential requirement for microglia to sense and interpret the brain microenvironment. This is crucial for microglia-synapse crosstalk that drives experience-dependent plasticity, a fundamental brain property impaired in several neuropsychiatric disorders. Phosphoproteomics profiling detects a large modulation of RhoGTPase signaling, predominantly of Rac1, in microglia of mice exposed to an environmental enrichment protocol known to induce experience-dependent brain plasticity and cognitive performance. Ablation of microglial Rac1 affects pathways involved in microglia-synapse communication, disrupts experience-dependent synaptic remodeling, and blocks the gains in learning, memory, and sociability induced by environmental enrichment. Our results reveal microglial Rac1 as a central regulator of pathways involved in the microglia-synapse crosstalk required for experience-dependent synaptic plasticity and cognitive performance.

Role of environmental enrichment on social interaction, anxiety, locomotion, and memory in Wistar rats under chronic methylphenidate intake.

Chronic use of various compounds can have long-lasting effects on animal behavior, and some of these effects can be influenced by the environment. Many environmental enrichment protocols have the potential to induce behavioral changes. The aim of the present study was to investigate how environmental enrichment can mitigate the effects of chronic methylphenidate consumption on the behavior of Wistar rats. The animals were housed for 20 days under either an environmental enrichment protocol (which included tubes of different shapes) or standard housing conditions. After seven days, half of the rats received 13 days of oral administration of methylphenidate (2 mg/kg). After seven days, the rats underwent behavioral tests, including the elevated plus maze (anxiety), open field (locomotion), object-in-place recognition test (spatial memory), and a test for social interaction (social behavior). The results showed that the enriched environmental condition reversed the enhanced time in the open arms of the elevated plus maze induced by methylphenidate (F[1,43] = 4.275, p = 0.045). Methylphenidate also enhanced exploratory rearing in the open field (F[1,43] = 4.663, p = 0.036) and the time spent in the open area of the open field (H[3] = 8.786, p = 0.032). The enriched environment mitigated the inhibition of social interaction with peers induced by methylphenidate (H[3] = 16.755, p < 0.001) as well as the preference for single exploratory behavior (H[3] = 9.041, p = 0.029). These findings suggest that environmental enrichment can counteract some of the effects of methylphenidate. These results are relevant for the clinical treatment of the long-lasting secondary effects associated with methylphenidate pharmacological treatment.

MSK1 is required for the beneficial synaptic and cognitive effects of enriched experience across the lifespan.

Positive experiences, such as social interaction, cognitive training and physical exercise, have been shown to ameliorate some of the harms to cognition associated with ageing. Animal models of positive interventions, commonly known as environmental enrichment, strongly influence neuronal morphology and synaptic function and enhance cognitive performance. While the profound structural and functional benefits of enrichment have been appreciated for decades, little is known as to how the environment influences neurons to respond and adapt to these positive sensory experiences. We show that adult and aged male wild-type mice that underwent a 10-week environmental enrichment protocol demonstrated improved performance in a variety of behavioural tasks, including those testing spatial working and spatial reference memory, and an enhancement in hippocampal LTP. Aged animals in particular benefitted from enrichment, performing spatial memory tasks at levels similar to healthy adult mice. Many of these benefits, including in gene expression, were absent in mice with a mutation in an enzyme, MSK1, which is activated by BDNF, a growth factor implicated in rodent and human cognition. We conclude that enrichment is beneficial across the lifespan and that MSK1 is required for the full extent of these experience-induced improvements of cognitive abilities, synaptic plasticity and gene expression.

Effect of environmental enrichment on relapse rates in patients with severe alcohol use disorder: protocol for a randomised controlled trial

IntroductionAlcohol use disorder (AUD) ranks among the most prevalent psychiatric disorders worldwide. Despite current treatments, more than half of patients relapse within weeks after treatment. In animal models, exposure to...

Environmental enrichment alleviates cognitive and psychomotor alterations and increases adult hippocampal neurogenesis in cocaine withdrawn mice.

Cocaine is a widely used psychostimulant drug whose repeated exposure induces persistent cognitive/emotional dysregulation, which could be a predictor of relapse in users. However, there is scarce evidence on effective treatments to alleviate these symptoms. Environmental enrichment (EE) has been shown to be associated with improved synaptic function and cellular plasticity changes related to adult hippocampal neurogenesis (AHN), resulting in cognitive enhancement. Therefore, EE could mitigate the negative impact of chronic administration of cocaine in mice and reduce the emotional and cognitive symptoms present during cocaine abstinence. In this study, mice were chronically administered with cocaine for 14 days, and control mice received saline. After the last cocaine or saline dose, mice were submitted to control or EE housing conditions, and they stayed undisturbed for 28 days. Subsequently, mice were evaluated with a battery of behavioural tests for exploratory activity, emotional behaviour, and cognitive performance. EE attenuated hyperlocomotion, induced anxiolytic-like behaviour and alleviated cognitive impairment in spatial memory in the cocaine-abstinent mice. The EE protocol notably upregulated AHN in both control and cocaine-treated mice, though cocaine slightly reduced the number of immature neurons. Altogether, these results demonstrate that EE could enhance hippocampal neuroplasticity ameliorating the behavioural and cognitive consequences of repeated administration of cocaine. Therefore, environmental stimulation may be a useful strategy in the treatment cocaine addiction.

Enrichment Protocol for Rat Models.

An environmental enrichment (EE) cage consisting of a broad living area and various stimulators triggers social, cognitive, and physical activities. EE has been utilized in a wide range of neurological and non-neurological studies. However, the details of the environmental enrichment protocol were not well described in these studies. This has resulted in uncertainty and inconsistency in methodology, which may thus fail to replicate environmental enrichment effects, influencing the study outcome. Here we describe the basic guidelines and present an easy-to-follow protocol for environmental enrichment in rat models. © 2021 Wiley Periodicals LLC. Basic Protocol: Environmental enrichment housing.

Effect of simple and complex enrichment added to standard-sized cages in behavioral, physiological, and neurological variables in female Swiss mice (Mus musculus).

Environmental enrichment (EE) has been a widely used tool to improve animal welfare, as well as to study brain plasticity. Traditional EE settings in the field of neuroscience employ highly complex cages with numerous objects and increased space, whereas more simple additions included for the control treatment are rarely considered in the experimental design. This leads to a lack of consistency of what neuroscientists designate as "standard housing," which might compromise the reproducibility of the results. Therefore, we employed standard-sized cages to study how different EE configurations can affect several biological markers of animal welfare. We first compared barren cages with cages containing nest material and a cardboard roll or cages having a complex set of elements. For this purpose, we studied anxiety-like behavior, corticosterone metabolites in feces, and cell survival in the hippocampus. Complex enrichment (CE) increased the concentration of corticosterone metabolites while also decreasing anxiety-like behavior. Interestingly, both simple and CEs were able to promote cell survival in the hippocampus, and this measure was positively correlated to corticosterone metabolites. Furthermore, in a second experiment, one of the elements of the CE was able to reduce anxiety-like behavior and blood glucose reactivity after exposure to a stressful situation. Altogether, this study calls attention about how sensitive experimental outcomes are to these simple EE elements. Even though EE is recommended by most guidelines for the care and use of laboratory animals, a detailed analysis of the EE protocol that is going to be implemented is highly encouraged. (PsycInfo Database Record (c) 2021 APA, all rights reserved).

Environmental Enrichment Potentiates Glucose-Induced Anorexia in Mice

The hypothalamus controls food intake and metabolism by integrating nutrient and hormonal signals from peripheral tissues. Both central and peripheral administration of glucose leads to a reduction in food intake in rodents. Similarly, administration of the adipocyte hormone leptin or the gastrointestinal hormone xenin reduces food intake. In contrast, impairments in hypothalamic signaling of these factors cause hyperphagia and obesity in rodents and humans. Environmental factors affect behavior including feeding behavior and energy metabolism in rodents and humans. Studies have found that environmental enrichment (EE), in which mice interact with complex sensory and motor stimulation, led to a significant reduction in adiposity and resistance to diet-induced obesity in mice. This effect is independent of energy expenditure and is associated with enhanced hypothalamic signaling, but the exact mechanism is unknown. We hypothesized that EE potentiates the feeding suppressing effects of anorectic signals. To address this hypothesis, 4-week-old male C57BL/6 mice were group housed (5/cage) under standard laboratory conditions or EE conditions with free access to regular rodent chow and feeding response to glucose, leptin and xenin was examined. EE cages were supplemented with a house, running wheels, igloos, wood logs, maze and nesting materials. Four weeks after initiating EE protocol, mice were fasted for 8 h and received an intraperitoneal injection of glucose (2 mg/g b.w.) or saline just before the onset of the dark phase. Treatment assignments were reversed for the second injection so that each animal received both treatments with a washout period of 1 week. Mice were given food immediately after the injection and food intake was measured for 4 h after the injection at 0.5–1 h intervals. The same design was repeated using leptin (2.5 μg/g b.w.) and xenin (15 or 50 μg/g b.w.). Glucose injection caused a significant reduction of food intake in both control and EE mice. However, anorexic effect of glucose was more significant in EE group compared to the control group (main effect of treatment: P = 0.0016 for control and P < 0.0001 for EE, two-way ANOVA). Significant reductions in food intake were observed between 0.5 and 2.5 h after glucose injection in EE mice, while no significant reduction was observed thereafter. Moreover, three-way ANOVA showed a significant interaction between housing condition and treatment (P = 0.0086). In contrast, although both leptin and xenin caused a significant reduction in food intake, there was no significant interaction between housing condition and treatment. These data suggest that environmental enrichment enhances the anorectic action of glucose without altering feeding response to leptin and xenin. It is speculated that enhanced hypothalamic glucose sensing may mediate beneficial effects of environmental enrichment on metabolism.

In aged rats, differences in spatial learning and memory influence the response to late-life Environmental Enrichment

It has clearly been demonstrated that cognitive stimulation, physical exercise, and social engagement help counteract age-related cognitive decline. However, several important issues remain to be addressed. Given the wide differences in cognitive impairment found among individuals of the same age, identifying the subjects who will benefit most from late-life interventions is one such issue. Environmental Enrichment (EE) is a particularly valuable approach to do this. In this study, aged (21-month-old) rats were assigned to a better (BL) or a worse (WL) learner group (training phase) and to a non-impaired (NI) or an impaired (I) group (probe phase) by their performance on the Morris Water Maze, using the test performances of adult (12-month-old) rats as the cut-offs. The aged rats were retested after a 12-week EE or standard housing (SH) protocol. After 12weeks, the performances of SH rats had deteriorated, whereas all rats benefited from EE, albeit in different ways. In particular, the animals assigned to the BL and the NI groups prior to EE still performed as well as the adult rats (performance preservation) whereas, critically, the animals assigned to the WL and the I groups before EE showed such improved performances that they reached the level of the adult rats (performance improvement), despite having aged further. EE seems to induce the preservation in BLs and the improvement in WLs of spatial search strategies and the preservation in NIs and the increase in Is of a focused and protract research of the escape point. Our findings suggest that late-life EE prevents spatial learning and memory decline in still cognitively preserved animals and stimulates residual functional reserve in already cognitively compromised animals. Future research should focus on individually tailored stimulation protocols to improve their effect and afford a better understanding of the underlying processes.

Simultaneous Monitoring of Wireless Electrophysiology and Memory Behavioral Test as a Tool to Study Hippocampal Neurogenesis.

Brainwaves amplitude obtained from electroencephalography (EEG) has been well-recognized as a basis for cognitive capacity, memory, and learning on animals and humans. Adult neurogenesis mechanism is also linked to memory and learning improvement. Traditionally, researchers used to assess learning and memory parameters in rodent models by behavioral tasks. Therefore, the simultaneous monitoring of behavioral changes and EEG is particularly interesting in correlating data between brain activity and task-related behaviors. However, most of the equipment required to perform both studies are either complex, expensive, or uses a wired setup network that hinders the natural animals' movement. In this study, EEG was recorded with a wireless electrophysiology device along with the execution of a novel object recognition task (NORT). The animal's behavior was monitored simultaneously by a video tracking system. Both recordings were analyzed offline by their timestamps which were synched to link EEG signals with the animal's actions. Subjects consist of adult Wistar rats after medium-term environmental enrichment treatment. Six skull screw electrodes were fixed in pairs on both hemispheres over frontal, central, and parietal regions and were referenced to an electrode located posterior of the nasal bone. NORT protocol consists of exposing the animal to two identical objects for 10 min. After 2 h and 24 h, one of the objects was replaced with a novel one. Exploration time for each object was monitored by a behavioral tracking software (BTS) and EEG data recording. The analysis of the EEG synced with behavioral data consists of estimations of alpha and beta relative band power and comparisons between novel object recognition versus familiar object exploration, between three experimental stages. In this manuscript, we have discussed electrodes manufacturing process, epidural electrodes implantation surgery, environmental enrichment protocol, NORT protocol, BTS setup, EEG - BTS coupling for simultaneous monitoring in real-time, and EEG data analysis based on automatic events detection.

Long-Term Effect of Environmental Enrichment on Reproductive Performance of Swiss Webster Mice and Their Female Offspring.

Simple SummaryThe aim of this study was to evaluate the effect of the implementation of an environmental enrichment (EE) plan on reproductive performance in mice, applied during one year period in Swiss Webster breeders and their female offspring used as recipients for embryo transfer. Birth rate, litter size, pups’ weight at weaning, interlitter interval and time to first litter were compared against a control group that did not receive EE. In mice breeders, EE significantly increased pups’ weight compared to control group (p < 0.01), without changing any other reproductive parameters. In the offspring females used as recipients, EE did not modify reproductive parameters. Our results demonstrate that this enrichment plan, which is usually applied in mouse facilities for animal well-being, enhances pup’s weight at weaning, with no negative effects on the reproductive performance of Swiss Webster mice.The aim of this study was to evaluate the effect of an environmental enrichment (EE) plan on the reproductive performance of Swiss Webster mice and their female offspring used as recipients for embryo transfer. A total of 54 breeder mice and 60 F1 females, used as foster mothers, were allocated in two experimental groups to receive or not receive EE for physical well-being. Reproductive outcomes of the Swiss trios such as birth rate and pup number, litter size, pups’ weight at weaning, interlitter interval and time to first litter were analyzed. Environmental enrichment significantly increased pups weight from breeding trios compared to the control group (14.4 ± 0.1 vs. 13.8g ± 0.1, EE vs. control, respectively; p < 0.01). Other parameters did not differ between both groups. Reproductive parameters of female offspring used as recipients for embryo transfer did not differ among groups subjected or not to EE. These data demonstrate that the EE protocol applied in Swiss Webster breeder mice positively enhanced pups weight, and did not interfere with other reproductive outcomes. In conclusion, this study supports the implementation of EE plans usually applied for animal welfare in mouse facilities with slight improvement in reproductive performance.

Early exposure to environmental enrichment protects male rats against neuropathic pain development after nerve injury

Because environmental elements modify chronic pain development and endogenous mechanisms of pain control are still a great therapeutic source, we investigated the effects of an early exposure to environmental enrichment (EE) in a translational model of neuropathic pain. Young male rats born and bred in an enriched environment, which did not count on running wheel, underwent chronic constriction injury (CCI) of sciatic nerve. EE abolished neuropathic pain behavior 14 days after CCI. Opioid receptors' antagonism reversed EE-analgesic effect. β-endorphin and met-enkephalin serum levels were increased only in EE-CCI group. Blockade of glucocorticoid receptors did not alter EE-analgesic effect, although corticosterone circulating levels were increased in EE animals. In the spinal cord, EE controlled CCI-induced serotonin increase. In DRG, EE blunted the expression of ATF-3 after CCI. Surprisingly, EE-CCI group showed a remarkable preservation of sciatic nerve fibers compared to NE-CCI group. This work demonstrated global effects induced by an EE protocol that explain, in part, the protective role of EE upon chronic noxious stimulation, reinforcing the importance of endogenous mechanisms in the prevention of chronic pain development.

Preconceptual paternal environmental stimulation alters behavioural phenotypes and adaptive responses intergenerationally in Swiss mice

Experimental research has recently revealed that paternal environmental conditions can influence the offspring phenotype through epigenetic mechanisms. However, it is unclear whether these effects impact adaptive responses in the offspring. Environmental enrichment (EE) is a well-established paradigm that promotes neural plasticity. We investigated whether EE in male mice could modify behaviours that are highly relevant for determining adaptive fitness, i.e. spatial memory, attractiveness and social dominance, in the offspring of outbred mice. Male Swiss mice were housed in either EE or standard housing from post-weaning to adulthood before breeding for offspring. Their offspring were raised in standard housing until adulthood then assessed for behavioural, physiological and molecular parameters. F0 male mice exposed to EE had lower body weight, higher adrenal, spleen and hippocampal weights, better novelty processing and spatial learning, greater hippocampal BDNF levels, and higher social dominance. Unexpectedly, their male offspring (F1) showed spatial memory impairment, lowered social dominance and were less attractive to receptive females, compared to controls. These ethologically relevant measures suggest a maladaptive response in the male F1 offspring. Interestingly, when separate cohorts of male F1 offspring of standard housing or EE fathers were exposed to 8-day EE protocol during adulthood, differences in spatial memory and attractiveness to receptive females were no longer observed between them. These results provide evidence that the paternal environment can influence the offspring’s adaptiveness.

Enriched Environment Procedures for Rodents: Creating a Standardized Protocol for Diverse Enrichment to Improve Consistency across Research Studies.

Exposure to environmental enrichment has beneficial effects on learning and memory, diverse neurobiological effects, and promotes recovery of function after brain injury. The effect of enrichment is produced by a combination of increased social interaction, physical activity, spatial complexity, and novelty. Procedures in the literature have, however, been idiosyncratic with poor consistency in the manner or extent to which protocols provide consistent enrichment. We provide an environmental enrichment protocol that can be easily replicated with minor details determined locally so that animals across cohorts and cages all experience a comparable level of enrichment. Procedures are outlined to generate and use a daily pool of suitably varied objects using a standardized format, with objects systematically varied up to a 40-day continuous period. Together with using a large group of rats in a suitably-sized cage, and regular shifting of the position of food and water and cage location, these procedures have produced robust effects in different laboratories and rat strain, thereby improving comparisons within and across laboratories. Non-enriched comparisons can vary, but typically would include grouped animals in standard laboratory housing without objects and with stable food and water locations. Enrichment is a safe non-pharamacological tool to examine behavioral and neurobiological processes in animal models of the lifespan, brain dysfunction and injury.

Anxiolytic and panicolytic-like effects of environmental enrichment seem to be modulated by serotonin neurons located in the dorsal subnucleus of the dorsal raphe

In a previous study, we showed that exposure of rats to a one-week environmental enrichment (EE) protocol decreases elevated T-maze (ETM) avoidance responses, an anxiolytic-like effect, without altering escape reactions, in clinical terms related to panic disorder. These anxiolytic-like effects were followed by decreased delta FosB-immunoreactivity (delta FosB-ir) in the cingulate cortex, dorsolateral and intermediate lateral septum, hippocampus (cornus of Ammon), anterior and dorsomedial hypothalamus, medial and basolateral amygdala and ventral region of the dorsal raphe nucleus. The purpose of the present study was to further investigate behavioral and neurophysiological alterations induced by EE exposure. For that, in a first experiment we verified if increasing the time of exposure to the same EE protocol used in our previous study (from one to two weeks) altered male Wistar rats’ ETM escape responses. All animals were tested in an open field, immediately after the ETM, for locomotor activity assessment. Since anxiety and panic-related reactions have been associated to the functioning of specific subnuclei of the dorsal raphe nucleus (DR), we also evaluated delta FosB-ir in serotonergic cells of DR regions. At last, we analyzed plasma corticosterone levels in animals submitted to EE and to standard housing. Results showed that a two-week exposure to EE decreases both ETM avoidance and escape reactions, inducing anxiolytic and panicolytic-like effects, respectively. There was also a significant decrease in the number of double staining neurons in the midrostral region of the dorsal subnucleus of the dorsal raphe. No changes in corticosterone levels, however, were observed. These results contribute to a better understanding of the effects of EE on anxiety and panic-related responses.

Aged rats with different performances at environmental enrichment onset display different modulation of habituation and aversive memory.

A wide agreement exists that environmental enrichment (EE) is most beneficial if introduced early in life, but numerous studies reported that also aged animals remain responsive. As age-related memory and cognition impairments are not uniform, an open question is whether EE might exert different effects in animals with different age-related deficits. A 12-week EE protocol was applied to late adult rats pretested for habituation and aversive memory. Animals were classified as low (LP) and high (HP) performers according to percent exploration change in Open Field test (OF) and as impaired (I) and not impaired (NI) according to latency in Step-through Passive Avoidance test (PA). Standard housing (SH) animals pretested by OF and PA, and naïve (non-pretested) EE and SH rats were used as controls. In comparison to pretest, after the housing protocol, EE LP ameliorated while EE HP and both SH HP and LP worsened their habituation pattern. The positive influence of EE on LP was probably due to the more active interaction with and the faster adaptation to surroundings promoted by continuous, multiple stimuli provided during the enriched housing. Regarding HP, EE did not boost the basal behavior, which likely represented the maximum achievable for that age, and the post housing exploration change dropped, as in SH animals, because of the retesting. After EE, a significant percentage of NI animals became I and a significant percentage of I animals became NI. The changes evidenced in the NI group likely depended on EE-related reduction of anxiety and the consequent more efficient coping with fearful situations. This hypothesis was strengthened by the observation that naïve EE animals were almost all I. Pretested EE I rats were not influenced by the rearing condition: their behavior was comparable to SH animals' behavior and determined by retesting. In conclusion, these results demonstrated that, when applied to aging rats, EE produces different effects based on pre-housing cognitive performances. The issue needs further analyses, but the observation that not all animals are able to take advantage of EE to the same extent suggests the opportunity to design individually tailored approaches to optimize their efficacy and minimize possible unwanted consequences.

Environmental Enrichment Results in Both Brain Connectivity Efficiency and Selective Improvement in Different Behavioral Tasks

Exposure to environmental enrichment (EE) has been a useful model for studying the effects of experience on brain plasticity, but to date, few is known about the impact of this condition on the brain functional networks that probably underlies the multiple behavioral improvements. Hence, we assessed the effect of an EE protocol in adult Wistar rats on the performance in several behavioral tasks testing different domains (Open field (OP): locomotor activity; Elevated-zero maze (EZM): anxiety-related behaviors; 5-choice serial reaction time task (5-CSRTT): attentional processes; 4-arm radial water maze (4-RAWM): spatial memory) in order to check its effectiveness in a wide range of functions. After this, we analyzed the functional brain connectivity underlying each experimental condition through cytochrome C oxidase (COx) histochemistry. Our EE protocol reduced both locomotor activity in the OP and anxiety-related behaviors in the EZM. On the other hand, enriched rats showed more accuracy in the 4-RAWM, whereas 5-CSRTT performance was not significantly ameliorated by EE condition. In relation to COx functional connectivity, we found that EE reduced the number of strong positive correlations both in basal and training conditions, suggesting a modulating effect on specific brain connections. Our results suggest that EE seems to have a selective effect on specific brain regions, such as prefrontal cortex and hippocampus, leading to a more efficient brain connectivity.

Distinct environmental enrichment protocols reduce anxiety but differentially modulate pain sensitivity in rats

Environmental enrichment (EE) can alter anxiety as well as perception of nociceptive stimuli, suggesting a relationship between well-being and analgesia. Considering that, we aimed to evaluate the influence of different EE types on anxiety and peripheral pain sensitivity of male Wistar rats. Animals were submitted to two different types of EE: On the first one, animals were housed in standard size cages after weaning and received three different objects, one type per week, on a regular basis (simple EE – sEE). On the second one, animals were born in an already enriched environment and, after five weeks, were housed in larger cages and received five different objects, three of each type per week, on a regular basis (improved EE – iEE). Control group remained in standard cages and did not receive objects. Within 7 weeks of life, anxiety and thermal sensibility were evaluated using elevated plus maze and tail flick tests, respectively. Mechanical hyperalgesia was analyzed in the presence of acute and chronic noxious stimuli by paw pressure test. Both EE protocols tested were effective in diminish anxiety but they did not alter thermal sensibility. On the other hand, sEE protocol did not alter acute and chronic induced mechanical hypersensitivity, whereas iEE completely abolished such pain behavior, even without exercise wheel as part of the enrichment. Our results show that specific parameters (anxiety and pain sensitivity) can be differentially modulated depending on EE protocol used, making possible the implementation of welfare to experimental animals in pain research.

Environmental enrichment attenuates the blood brain barrier dysfunction induced by the neonatal hypoxia-ischemia

Environmental enrichment (EE) is considered an efficient neuroprotector against neonatal hypoxia-ischemia (HI). Nevertheless, the mechanisms involved are not yet clear. In this context, the aim of this study was to investigate the effects of neonatal HI and environmental stimulation in the hippocampus of rats at 3 different time points (PND 8, 22 and 60), evaluating some aspects of BBB structure and function. Seven-day-old Wistar rats were divided into four groups: a control group maintained in a standard environment (CTSE), a control group maintained in an enrichment environment (CTEE), an HI group maintained in a standard environment (HISE) and an HI group maintained in an enrichment environment (HIEE). At the 7th postnatal day (PND), rats were submitted to the Levine-Rice model of neonatal HI. This method consists of permanent occlusion of the right common carotid artery with subsequent exposure to hypoxia. Rats from CTEE and HIEE were stimulated with environmental enrichment. The EE protocol started 24h after HI, in which pup rats with their dams were stimulated in a maintained EE (PND 8–22). Subsequently, animals were submitted to daily EE (1h/day, PND 23–60). The expression of some proteins involved in BBB structure (β-catenin, occludin, connexin-43, aquaporin-4, glut-1 and GFAP) were quantified by western blotting in the hippocampi of rats in three periods, at PND 8, 22 and 60. The BBB permeability and integrity was assessed by Evans blue staining and the immunohistochemistry for GFAP in the CA1 region of the hippocampus were also performed. The results showed an HI-induced decreased occludin expression at PND 22 and low levels of occludin, β-catenin and GFAP at PND 60 in the hippocampus of the hypoxic-ischemic rats. Interestingly, in young and adult rats, EE reversed these effects. Evans blue extravasation into the brain parenchyma confirmed the BBB dysfunction brought on by HI. No differences were observed at PND 8, probably due to the immaturity of the BBB at this age. The present study makes an important contribution to understanding the mechanism of the hypoxic-ischemic brain damage and also to presents, for the first time, the recovery of BBB dysfunction as a possible pathway for the protective effect of EE.

Sex differences after environmental enrichment and physical exercise in rats when solving a navigation task.

The effects of early environmental enrichment (EE) and voluntary wheel running on the preference for using a landmark or pool geometry when solving a simple spatial task in adult male and female rats were assessed. After weaning, rats were housed in same-sex pairs in enriched or standard cages (EE and control groups) for two and a half months. Then the rats were trained in a triangular-shaped pool to find a hidden platform whose location was defined in terms of these two sources of information, a landmark outside the pool and a particular corner of the pool. As expected, enriched rats reached the platform faster than control animals, and males and females did not differ. Enriched rats also performed better on subsequent test trials without the platform with the cues individually presented (either pool geometry or landmark). However, on a preference test without the platform, a clear sex difference was found: Females spent more time in an area of the pool that corresponded to the landmark, whereas males spent more time in the distinctive corner of the pool. The present EE protocol did not alter females' preference for the landmark cue. The results agree with the claim that environmental enrichment is a consequence of a reduced anxiety response (measured by thigmotaxis) during cognitive testing. A possible implication of ancestral selection pressures is discussed.