Environmental Impoverishment, Aging, and Reduction in Mastication Affect Mouse Innate Repertoire to Explore Novel Environments and to Assess Risk

Studies indicate that inhibition of adequate masticatory function, due to soft diet, occlusal disharmony, or molar losses affects the cognitive behavior of rodents. However, no study has tested the effects on new environments exploration and risk assessment coupled with a combination of masticatory function rehabilitation and environmental enrichment. In the present report, we tested the hypothesis that age, environment, and masticatory changes may interact and alter exploratory patterns of locomotor activity and mice preferences in an open field (OF) arena. As OF arenas are widely used to measure anxiety-like behavior in rats and mice. We examined in an open arena, the exploratory and locomotor activities of mature (6-month-old; 6M), late mature (12-month-old; 12M), and aged (18-month-old; 18M) mice, subjected to distinct masticatory regimens and environments. Three different regimens of masticatory activity were used: continuous normal mastication with hard pellets (HD); normal mastication followed by reduced mastication with equal periods of pellets followed by soft powder – HD/SD; or rehabilitated masticatory activity with equal periods of HD, followed by powder, followed by pellets – HD/SD/HD). Under each diet regimen, half of the individuals were raised in standard cages [impoverished environment (IE)] and the other half in enriched cages [enriched environment (EE)]. Animals behavior on the open field (OF) task were recorded by webcam and analyzed with Any Maze software (Stöelting). The locomotor and exploratory activities in OF task declined with age, and this was particularly evident in 18M HD EE mice. Although all groups kept their preference by the peripheral zone, the outcomes were significantly influenced by interactions between environment, age, and diet. Independent of diet regime, 6M young mice maintained in an EE where voluntary exercise apparatus is available, revealed significant less body weight than all other groups. Although body weight differences were minimized as age progressed, 18M EE group revealed intragroup significant influence of diet regimens. We suggest that long life environmental enrichment reduces the tendency to avoid open/lit spaces (OF) and this is particularly influenced by masticatory activity. These measurements may be useful in discussions of anxiety-related tasks

Spatial memory decline after masticatory deprivation and aging is associated with altered laminar distribution of CA1 astrocytes

<p>Abstract</p> <p>Background</p> <p>Chewing imbalances are associated with neurodegeneration and are risk factors for senile dementia in humans and memory deficits in experimental animals. We investigated the impact of long-term reduced mastication on spatial memory in young, mature and aged female albino Swiss mice by stereological analysis of the laminar distribution of CA1 astrocytes. A soft diet (SD) was used to reduce mastication in the experimental group, whereas the control group was fed a hard diet (HD). Assays were performed in 3-, 6- and 18-month-old SD and HD mice.</p> <p>Results</p> <p>Eating a SD variably affected the number of astrocytes in the CA1 hippocampal field, and SD mice performed worse on water maze memory tests than HD mice. Three-month-old mice in both groups could remember/find a hidden platform in the water maze. However, 6-month-old SD mice, but not HD mice, exhibited significant spatial memory dysfunction. Both SD and HD 18-month-old mice showed spatial memory decline. Older SD mice had astrocyte hyperplasia in the strata pyramidale and oriens compared to 6-month-old mice. Aging induced astrocyte hypoplasia at 18 months in the lacunosum-moleculare layer of HD mice.</p> <p>Conclusions</p> <p>Taken together, these results suggest that the impaired spatial learning and memory induced by masticatory deprivation and aging may be associated with altered astrocyte laminar distribution and number in the CA1 hippocampal field. The underlying molecular mechanisms are unknown and merit further investigation.</p

Enriched environment and masticatory activity rehabilitation recover spatial memory decline in aged mice

BACKGROUND: To measure the impact of masticatory reduction on learning and memory, previous studies have produced experimental masticatory reduction by modified diet or molar removal. Here we induced spatial learning impairment in mice by reducing masticatory activity and then tested the effect of a combination of environmental enrichment and masticatory rehabilitation in recovering spatial learning at adulthood and in later life. For 6 months (6M) or 18 months (18M), we fed three groups of mice from postnatal day 21 respectively with a hard diet (HD) of pellets; pellets followed by a powdered, soft diet (HD/SD, divided into equal periods); or pellets followed by powder, followed by pellets again (HD/SD/HD, divided into equal periods). To mimic sedentary or active lifestyles, half of the animals from each group were raised from weaning in standard cages (impoverished environment; IE) and the other half in enriched cages (enriched environment; EE). To evaluate spatial learning, we used the Morris water maze. RESULTS: IE6M-HD/SD mice showed lower learning rates compared with control (IE6M-HD) or masticatory rehabilitated (IE6MHD/SD/HD) animals. Similarly, EE-HD/SD mice independent of age showed lower performance than controls (EE-HD) or rehabilitated mice (EE-HD/SD/HD). However, combined rehabilitation and EE in aged mice improved learning rate up to control levels. Learning rates did not correlate with swim speed. CONCLUSIONS: Reduction in masticatory activity imposed on mice previously fed a hard diet (HD/SD) impaired spatial learning in the Morris water maze. In adults, masticatory rehabilitation recovered spatial abilities in both sedentary and active mice, and rehabilitation of masticatory activity combined with EE recovered these losses in aged mice

Influência da atividade mastigatória e do ambiente sobre o aprendizado espacial e o padrão da atividade exploratória em modelo murinho senil

Para medir possíveis influências da mastigação e do estilo de vida sedentário em modelo murino senil, impusemos um de três regimes de dieta aos diferentes grupos experimentais do 21º dia pós-natal até 6 ou 18 meses de vida: dieta sólida tipo pellet; dieta em pellet seguida por uma em pó, farelada; ou dieta peletizada seguida de pó e novamente pellet, com intervalos de tempo iguais em cada dieta. Para mimetizar o estilo de vida sedentário ou ativo, os animais foram criados, respectivamente, em gaiolas-padrão (ambiente empobrecido-AP) ou em gaiolas enriquecidas (ambiente enriquecido-AE). Para medir os efeitos da dieta, do ambiente e da idade sobre a atividade exploratória, realizamos o teste do campo aberto, onde camundongos jovens de AP que sofreram alteração da atividade mastigatória demonstraram maior preferência pela zona periférica, mas no envelhecimento e no AE essas diferenças foram minimizadas. Nos velhos de AE, essas diferenças reapareceram. Já sobre as influências na aprendizagem e memória espacial, aplicamos o labirinto aquático de Morris e vimos que a redução da atividade mastigatória, independente do ambiente, diminuiu a taxa média de aprendizado espacial e sua reabilitação recuperou as perdas associadas em animais jovens e, quando combinada ao AE, melhorou a taxa de aprendizado em velhos. Não se encontrou correlação entre taxa de aprendizado e velocidade de nado dos camundongos sugerindo que os déficits são de natureza cognitiva. Concluímos assim, que a alteração da atividade mastigatória influencia o padrão de exploração por zonas no campo aberto e a estimulação ambiental acentua os seus efeitos no envelhecimento, privilegiando a preferência pela zona periférica e a redução da atividade mastigatória prejudica a memória espacial durante o teste do labirinto aquático de Morris e a sua reabilitação é capaz de recuperar as habilidades espaciais, mas em idosos é necessária a combinação com um AE.ABSTRACT: To measure possible influences of the mastication and sedentary lifestyle, we imposed masticatory patterns with three different experimental diet regimes starting at 21^st postnatal day during 6 (6M) or 18 (18M) months: continuous pellet hard diet, equal periods of hard followed by soft diet or equal periods of hard followed by soft followed by hard diet. To mimic the sedentary and active lifestyles, the animals were raised, respectively in standard (AP) or enriched cages (AE). To measure the effects of diet, environment and age on the exploratory activity, we performed the open field test. Young mice (AP) with altered masticatory activity demonstrated more preference for peripheric zone, but in aging and AE animals these differences were minimized. In the aging specimens maintained in AE, these differences were reappeared. About influences on learning and spatial memory, we apply the Morris water maze and the reduced masticatory activity, regardless of the environment, decreased the average rate of spatial learning and rehabilitation recovered the losses associated in young animals and the association with AE improved the learning rate in old mice. No significant correlations were observed between swimming speed and learning rate. We conclude that the change in masticatory activity influences the pattern of exploration by zones in the open field and environmental stimulation enhances the effects of aging, emphasizing the preference for peripheric zone and reduced masticatory activity impairs spatial memory during the test of the Morris water maze and rehabilitation is able to recover spatial ability. Here, the combination with AE is required in aging

Sedentary life and reduced mastication impair spatial learning and memory and differentially affect dentate gyrus astrocyte subtypes in the aged mice

Brazilian Research Council CNPq (grant number 475677/2008-0) Fundação Amazônia Paraense de Amparo à Pesquisa (FAPESPA, grant number 136/08). Fundação de Amparo e Desenvolvimento da Pesquisa (FADESP) and the Pró-Reitoria de Pesquisa e Pós-Graduação (PROPESP/UFPA) Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES). Programa Nacional de Cooperação Acadêmica da Amazônia – DRI (Processo CAPES No. 88881.370785/2019-01). Fundação para a Ciência e a Tecnologia (PTDC/MED-NEU/31395/2017, LISBOA-01-0145-FEDER031395, and UID/DTP/04138/2018-2021), Santa Casa da Misericórdia (ALS Research Grant).Universidade Federal do Pará. Instituto de Ciências Biológicas. Hospital Universitário João de Barros Barreto. Laboratório de Investigações em Neurodegeneração e Infecção. Belém, PA, Brazil / Centro Universitário do Estado do Pará. Curso de Medicina. Belém, PA, Brazil.Universidade Federal do Pará. Instituto de Ciências Biológicas. Hospital Universitário João de Barros Barreto. Laboratório de Investigações em Neurodegeneração e Infecção. Belém, PA, Brazil.Universidade Federal do Pará. Instituto de Ciências Biológicas. Hospital Universitário João de Barros Barreto. Laboratório de Investigações em Neurodegeneração e Infecção. Belém, PA, Brazil / Ministério da Saúde. Secretaria de Vigilância em Saúde. Instituto Evandro Chagas. Laboratório de Microscopia Eletrônica. Ananindeua, PA, Brasil.University of Oxford. Department of Pharmacology. Laboratory of Experimental Neuropathology. Oxford, United Kingdom.Universidade de Lisboa. Faculty of Pharmacy. Research Institute for Medicines. Lisbon, Portugal / Universidade de Lisboa. Faculty of Pharmacy. Department of Biochemistry and Human Biology. Lisbon, Portugal.Universidade Federal do Pará. Instituto de Ciências Biológicas. Hospital Universitário João de Barros Barreto. Laboratório de Investigações em Neurodegeneração e Infecção. Belém, PA, Brazil.Universidade Federal do Pará. Instituto de Ciências Biológicas. Hospital Universitário João de Barros Barreto. Laboratório de Investigações em Neurodegeneração e Infecção. Belém, PA, Brazil.To explore the impact of reduced mastication and a sedentary lifestyle on spatial learning and memory in the aged mice, as well as on the morphology of astrocytes in the molecular layer of dentate gyrus (MolDG), different masticatory regimens were imposed. Control mice received a pellet-type hard diet, while the reduced masticatory activity group received a pellet diet followed by a powdered diet, and the masticatory rehabilitation group received a pellet diet, followed by powder diet and then a pellet again. To mimic sedentary or active lifestyles, mice were housed in an impoverished environment of standard cages or in an enriched environment. The Morris Water Maze (MWM) test showed that masticatory-deprived group, regardless of environment, was not able to learn and remember the hidden platform location, but masticatory rehabilitation combined with enriched environment recovered such disabilities. Microscopic three-dimensional reconstructions of 1,800 glial fibrillary acidic protein (GFAP)-immunolabeled astrocytes from the external third of the MolDG were generated using a stereological systematic and random sampling approach. Hierarchical cluster analysis allowed the characterization into two main groups of astrocytes with greater and lower morphological complexities, respectively, AST1 and AST2. When compared to the hard diet group subjected to impoverished environment, deprived animals maintained in the same environment for 6 months showed remarkable shrinkage of astrocyte branches. However, the long-term environmental enrichment (18-monthold) applied to the deprived group reversed the shrinkage effect, with significant increase in the morphological complexity of AST1 and AST2, when in an impoverished or enriched environment. During housing under enriched environment, complexity of branches of AST1 and AST2 was reduced by the powder diet (pellet followed by powder regimes) in young but not in old mice, where it was reversed by pellet diet (pellet followed by powder and pellet regime again). The same was not true for mice housed under impoverished environment. Interestingly, we were unable to find any correlation between MWM data and astrocyte morphological changes. Our findings indicate that both young and aged mice subjected to environmental enrichment, and under normal or rehabilitated masticatory activity, preserve spatial learning and memory. Nonetheless, data suggest that an impoverished environment and reduced mastication synergize to aggravate age-related cognitive decline; however, the association with morphological diversity of AST1 and AST2 at the MolDG requires further investigation

Comparative analysis between Open Field and Elevated Plus Maze tests as a method for evaluating anxiety-like behavior in mice

Anxiety is being increasingly diagnosed in the elderly population. In this sense, epidemiologic data have linked late-life anxiety disorders to increased cognitive decline, morbidity, and even mortality. In addition, studies have already reported the influence of the environment on the association between aging and anxiety. Therefore, the present study aimed to conduct a comparative analysis between Elevated Plus Maze (EPM) and Open Field (OF) tests as methods for evaluating mice's anxiety-like behavior, considering environmental and age variables. For this, eighty Female albino Swiss mice aged 6, 12, and 18 months were housed in an impoverished environment (IE) and enriched environment (EE). Following this, the animals were tested in EPM and OF tests. The environment and age affect the anxiety-like behavior of the mice in the OF, with a difference between the animals of 6 and 18 months, only in the EE (p < 0.021). However, in the EPM, it does not occur. Despite that, the environment affected the distance traveled by the mice in the EPM, where the IE animals showed greater exploratory activity than the EE, only in the 18-month group (p < 0.001). No environmental influences were detected in the OF. Concerning age, in the EPM, animals in the 18-month-old group traveled shorter distances compared to the 6-month group (p < 0.001) and the 12-month group (p < 0.001), only in EE. In turn, in the OF there was a decrease in the distance traveled in the 18-month group compared to the 6-month group (p = 0.012), only in the IE. Thus, the divergences between the results of EPM and OF instigate a better evaluation of the parameters analyzed in each test

The Sedentary Lifestyle and Masticatory Dysfunction: Time to Review the Contribution to Age-Associated Cognitive Decline and Astrocyte Morphotypes in the Dentate Gyrus

As aging and cognitive decline progresses, the impact of a sedentary lifestyle on the appearance of environment-dependent cellular morphologies in the brain becomes more apparent. Sedentary living is also associated with poor oral health, which is known to correlate with the rate of cognitive decline. Here, we will review the evidence for the interplay between mastication and environmental enrichment and assess the impact of each on the structure of the brain. In previous studies, we explored the relationship between behavior and the morphological features of dentate gyrus glial fibrillary acidic protein (GFAP)-positive astrocytes during aging in contrasting environments and in the context of induced masticatory dysfunction. Hierarchical cluster and discriminant analysis of GFAP-positive astrocytes from the dentate gyrus molecular layer revealed that the proportion of AST1 (astrocyte arbors with greater complexity phenotype) and AST2 (lower complexity) are differentially affected by environment, aging and masticatory dysfunction, but the relationship is not straightforward. Here we re-evaluated our previous reconstructions by comparing dorsal and ventral astrocyte morphologies in the dentate gyrus, and we found that morphological complexity was the variable that contributed most to cluster formation across the experimental groups. In general, reducing masticatory activity increases astrocyte morphological complexity, and the effect is most marked in the ventral dentate gyrus, whereas the effect of environment was more marked in the dorsal dentate gyrus. All morphotypes retained their basic structural organization in intact tissue, suggesting that they are subtypes with a non-proliferative astrocyte profile. In summary, the increased complexity of astrocytes in situations where neuronal loss and behavioral deficits are present is counterintuitive, but highlights the need to better understand the role of the astrocyte in these conditions

The Masticatory Activity Interference in Quantitative Estimation of CA1, CA3 and Dentate Gyrus Hippocampal Astrocytes of Aged Murine Models and under Environmental Stimulation

Studies indicating the influence of masticatory dysfunction, due to a soft diet or lack of molars, on impairing spatial memory and learning have led to research about neuronal connections between areas and cell populations possibly affected. In this sense, with scarce detailed data on the subfields of hippocampus in dementia neurodegeneration, there is no information about astrocytic responses in its different layers. Thus, considering this context, the present study evaluated the effects of deprivation and rehabilitation of masticatory activity, aging, and environmental enrichment on the stereological quantification of hippocampal astrocytes from layers CA1, CA3, and DG. For this purpose, we examined mature (6-month-old; 6M), and aged (18-month-old; 18M) mice, subjected to distinct masticatory regimens and environments. Three different regimens of masticatory activity were applied: continuous normal mastication with hard pellets (HD); normal mastication followed by deprived mastication with equal periods of pellets followed by soft powder (HD/SD); or rehabilitated masticatory activity with equal periods of HD, followed by powder, followed by pellets (HD/SD/HD). Under each specific regimen, half of the animals were raised in standard cages (impoverished environment (IE)) and the other half in enriched cages (enriched environment (EE)), mimicking sedentary or active lifestyles. Microscopic stereological, systematic, and random sampling approaches with an optical dissector of GFAP-immunolabeled astrocytes were done, allowing for an astrocyte numerical estimate. Stratum moleculare and hilus, from the dentate gyrus (DG) and Strata Lacunosum-Moleculare, Oriens, and Radiatum, similarly to the dentate gyrus, showed no significant change in any of the investigated variables (age, diet, or environment) in these layers. However, in Stratum radiatum, it was possible to observe significant differences associated with diet regimens and age. Therefore, diet-related differences were found when the HD 18M IE group was compared to the HD/SD/HD 18-month-old group in the same environment (IE) (p = 0.007). In the present study, we present modulatory factors (masticatory function, environmental enrichment, and aging) for the differentiated quantitative laminar response in the hippocampal regions, suggesting other studies to read the plasticity and responsiveness of astrocytes, including the molecular background

NEOTROPICAL CARNIVORES: a data set on carnivore distribution in the Neotropics

Mammalian carnivores are considered a key group in maintaining ecological health and can indicate potential ecological integrity in landscapes where they occur. Carnivores also hold high conservation value and their habitat requirements can guide management and conservation plans. The order Carnivora has 84 species from 8 families in the Neotropical region: Canidae; Felidae; Mephitidae; Mustelidae; Otariidae; Phocidae; Procyonidae; and Ursidae. Herein, we include published and unpublished data on native terrestrial Neotropical carnivores (Canidae; Felidae; Mephitidae; Mustelidae; Procyonidae; and Ursidae). NEOTROPICAL CARNIVORES is a publicly available data set that includes 99,605 data entries from 35,511 unique georeferenced coordinates. Detection/non-detection and quantitative data were obtained from 1818 to 2018 by researchers, governmental agencies, non-governmental organizations, and private consultants. Data were collected using several methods including camera trapping, museum collections, roadkill, line transect, and opportunistic records. Literature (peer-reviewed and grey literature) from Portuguese, Spanish and English were incorporated in this compilation. Most of the data set consists of detection data entries (n = 79,343; 79.7%) but also includes non-detection data (n = 20,262; 20.3%). Of those, 43.3% also include count data (n = 43,151). The information available in NEOTROPICAL CARNIVORES will contribute to macroecological, ecological, and conservation questions in multiple spatio-temporal perspectives. As carnivores play key roles in trophic interactions, a better understanding of their distribution and habitat requirements are essential to establish conservation management plans and safeguard the future ecological health of Neotropical ecosystems. Our data paper, combined with other large-scale data sets, has great potential to clarify species distribution and related ecological processes within the Neotropics. There are no copyright restrictions and no restriction for using data from this data paper, as long as the data paper is cited as the source of the information used. We also request that users inform us of how they intend to use the data