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There is still limited mechanistic insight into how the interaction of individuals with their environment results in the emergence of individuality in behavior and brain structure. Nevertheless, the idea that personal activity shapes the brain is implicit in strategies for healthy cognitive aging as well as in the idea that individuality is reflected in the brain's connectome. We have shown that even isogenic mice kept in a shared enriched environment (ENR) developed divergent and stable social and exploratory trajectories. As these trajectories-measured as roaming entropy (RE)-positively correlated with adult hippocampal neurogenesis, we hypothesized that a feedback between behavioral activity and adult hippocampal neurogenesis might be a causal factor in brain individualization. We used cyclin D2 knockout mice with constitutively extremely low levels of adult hippocampal neurogenesis and their wild-type littermates. We housed them for 3 months in a novel ENR paradigm, consisting of 70 connected cages equipped with radio frequency identification antennae for longitudinal tracking. Cognitive performance was evaluated in the Morris Water Maze task (MWM). With immunohistochemistry we confirmed that adult neurogenesis correlated with RE in both genotypes and that D2 knockout mice had the expected impaired performance in the reversal phase of the MWM. But whereas the wild-type animals developed stable exploratory trajectories with increasing variance, correlating with adult neurogenesis, this individualizing phenotype was absent in D2 knockout mice. Here the behaviors started out more random and revealed less habituation and low variance. Together, these findings suggest that adult neurogenesis contributes to experience-dependent brain individualization.

Personalized medicine intensifies interest in experimental paradigms that delineate sources of phenotypic variation. The paradigm of environmental enrichment allows for comparisons among differently housed laboratory rodents to unravel environmental effects on brain plasticity and related phenotypes. We have developed a new longitudinal variant of this paradigm, which allows to investigate the emergence of individuality, the divergence of individual behavioral trajectories under a constant genetic background and in a shared environment. We here describe this novel method, the 'Individuality Paradigm,' which allows to investigate mechanisms that drive individuality. Various aspects of individual activity are tracked over time to identify the contribution of the non-shared environment, that is the extent to which the experience of an environment differs between individual members of a population. We describe the design of this paradigm in detail, lay out its scientific potential beyond the published studies and discuss how it differs from other approaches to study individuality. The custom-built cage system, commercially marketed as 'ColonyRack', allows mice to roam freely between 70 cages through connector tubes equipped with ring antennas that detect each animal's ID from an RFID transponder implanted in the animal's neck. The system has a total floor area of 2.74 m2 and its spatial resolution corresponds to the size of the individual cages. Spatiotemporally resolved antenna contacts yield longitudinal measures of individual behavior, including the powerful measure of roaming entropy (RE). The Individuality Paradigm provides a rodent model of the making of individuality and the impact of the 'non-shared' environment on life-course development.

Panic disorder can be categorized into the nonrespiratory or the respiratory subtypes, the latter comprising dyspnea, shortness of breath, chest pain, feelings of suffocation, and paresthesias. Doxapram is an analeptic capable of inducing panic attacks with respiratory symptoms in individuals diagnosed with the disorder; however, its neuroanatomical targets and its effects on experimental animals remain uncharacterized. One of the brain regions proposed to trigger panic attacks is the midbrain periaqueductal gray (PAG). Therefore, in this study, we evaluated the effects of doxapram in Fos (c-Fos) protein expression in the PAG and characterized its cardiorespiratory and behavioral effects on the elevated T maze and in the conditioned place aversion (CPA) paradigms. Doxapram increased Fos expression in different columns of the PAG, increased respiratory frequency, decreased heart rate, and increased arterial pressure when injected via intravenous route. Alprazolam, a panicolytic benzodiazepine, injected via intraperitoneal route, decreased respiratory frequency, whereas URB597, an anandamide hydrolysis inhibitor injected via intraperitoneal route, was ineffective. Doxapram injected via intraperitoneal route induced an anxiogenic-like effect in the elevated T-maze model; however, it failed to induce CPA. This study suggests that the cardiorespiratory and behavioral effects of doxapram in rodents serve as an experimental model that can provide insights into the neurobiology of panic attacks.

Background: Evidence has revealed an association between familial hypercholesterolemia and cognitive impairment. In this regard, a connection between cognitive deficits and hippocampal blood-brain barrier (BBB) breakdown was found in low-density lipoprotein receptor knockout mice (LDLr–/–), a mouse model of familial hypercholesterolemia. Objective: Herein we investigated the impact of a hypercholesterolemic diet on cognition and BBB function in C57BL/6 wild-type and LDLr–/–mice. Methods: Animals were fed with normal or high cholesterol diets for 30 days. Thus, wild-type and LDLr–/–mice were submitted to memory paradigms. Additionally, BBB integrity was evaluated in the mice’s prefrontal cortices and hippocampi. Results: A tenfold elevation in plasma cholesterol levels of LDLr–/–mice was observed after a hypercholesterolemic diet, while in wild-type mice, the hypercholesterolemic diet exposure increased plasma cholesterol levels only moderately and did not induce cognitive impairment. LDLr–/–mice presented memory impairment regardless of the diet. We observed BBB disruption as an increased permeability to sodium fluorescein in the prefrontal cortices and hippocampi and a decrease on hippocampal claudin-5 and occludin mRNA levels in both wild-type and LDLr–/–mice treated with a hypercholesterolemic diet. The LDLr–/–mice fed with a regular diet already presented BBB dysfunction. The BBB-increased leakage in the hippocampi of LDLr–/–mice was related to high microvessel content and intense astrogliosis, which did not occur in the control mice. Conclusion: Therefore, LDLr–/–mice seem to be more susceptible to cognitive impairments and BBB damage induced by exposure to a high cholesterol diet. Finally, BBB disruption appears to be a relevant event in hypercholesterolemia-induced brain alterations.

Experience-induced differences in behavior, brain plasticity, and epigenetics persist even after stimulus withdrawal., Individuals differ in their response to environmental stimuli, but the stability of individualized behaviors and their associated changes in brain plasticity are poorly understood. We developed a novel model of enriched environment to longitudinally monitor 40 inbred mice exploring 35 connected cages over periods of 3 to 6 months. We show that behavioral individuality that emerged during the first 3 months of environmental enrichment persisted when mice were withdrawn from the enriched environment for 3 additional months. Behavioral trajectories were associated with stable interindividual differences in adult hippocampal neurogenesis and persistent epigenetic effects on neuronal plasticity genes in the hippocampus. Using genome-wide DNA methylation sequencing, we show that one-third of the DNA methylation changes were maintained after withdrawal from the enriched environment. Our results suggest that, even under conditions that control genetic background and shared environment, early-life experiences result in lasting individualized changes in behavior, brain plasticity, and epigenetics.

Individuals differ in their response to environmental stimuli, but the stability of individualized behaviors and their associated changes in brain plasticity are poorly understood. We developed a novel model of enriched environment to longitudinally monitor 40 inbred mice exploring 35 connected cages over periods of three to six months. We show that behavioral individuality that emerged during the first three months of environmental enrichment persisted when mice were withdrawn from the enriched environment for three additional months. Behavioral trajectories were associated with stable inter-individual differences in adult hippocampal neurogenesis and persistent epigenetic effects on neuronal plasticity genes in the hippocampus. Using genome-wide DNA methylation sequencing, we show that one third of the DNA methylation changes were maintained after withdrawal from the enriched environment. Our results suggest that, even under the most constraint conditions controlling genes and environment, early-life experiences result in lasting individualized changes in behavior and brain plasticity.

Cocaine is a psychostimulant drug that facilitates monoaminergic neurotransmission. The endocannabinoid system, comprising the cannabinoid receptors (CB1R and CB2R), the endocannabinoids, and their metabolizing-enzymes, modulates the mesolimbic dopaminergic pathway and represents a potential target for the treatment of addiction. Here, we tested the hypothesis that the cannabinoid receptors are implicated in cocaine-induced motor sensitization, conditioned place preference (CPP), and hippocampal activation. Male Swiss mice received injections of AM251 (CB1R antagonist; 0.3–10 mg/kg) or JWH133 (CB2R agonist; 1–10 mg/kg) before acquisition or expression of cocaine (20 mg/kg)-induced sensitization and CPP. After the CPP test, cFos-staining was employed as a marker of neuronal activation in the hippocampus. AM251 inhibited the acquisition (0.3, 1, and 3 mg/kg) and expression (1 and 3 mg/kg) of sensitization, as well as the acquisition (10 mg/kg) of CPP. JWH133 inhibited the acquisition (0.3 and 1 mg/kg) and expression (1 and 3 mg/kg) of both sensitization and CPP. JWH133 effects were reversed by AM630 (CB2R antagonist; 5 mg/kg). AM251 and JWH133 also prevented neuronal activation (c-Fos expression) in the hippocampus of CPP-exposed animals. CB1R and CB2R have opposite roles in modulating cocaine-induced sensitization and CPP, possibly by preventing neuronal activation in the hippocampus.

CNPq - Conselho Nacional de Desenvolvimento Científico e Tecnológico FAPEMIG - Fundação de Amparo à Pesquisa do Estado de Minas Gerais CAPES - Coordenação de Aperfeiçoamento de Pessoal de Nível Superior As drogas de abuso, a exemplo da cocaína, exercem efeito recompensador e facilitam a neurotransmissão dopaminérgica na via mesolímbica. O sistema endocanabinoide, representado por receptores canabinoides (CB1 e CB2), endocanabinoides e as enzimas que os sintetizam e degradam, modula esta via de neurotransmissão e pode ser um importante alvo de terapias farmacológicas para o tratamento do transtorno por uso de substância. Neste trabalho, testamos a hipótese de que receptores canabinoides estão evolvidos nos efeitos de sensibilização motora, preferência condicionada ao lugar (PCL) e ativação de neurônios hipocampais induzidas por cocaína. Para tal, camundongos Swiss receberam injeções de AM251 (antagonista de receptor CB1) ou JWH133 (agonista de receptor CB2) durante as fases de aquisição ou expressão dos comportamentos citados. O AM251 inibiu a sensibilização, quando administrado nas fases de aquisição (0,3, 1 e 3 mg/kg) ou expressão (1 e 3 mg/kg), e a PCL, quando administrado na fase de aquisição (10 mg/kg). O JWH133 não impediu o efeito agudo da cocaína. Porém, atenuou a sensibilização quando administrado nas fases de aquisição (0,3 e 1 mg/kg) ou expressão (1 e 3 mg/kg). Além disso, a administração de JWH133 (10 mg/kg) preveniu a PCL por cocaína em ambas as fases. Os efeitos do JWH133 foram revertidos com a administração de AM630 (antagonista de receptores CB2). A atenuação da memória contextual induzida por cocaína foi acompanhada por diminuição na ativação neuronial (células positivas para c-Fos) no hipocampo dos animais tratados com AM251 e JWH133. Assim, os efeitos comportamentais e celulares induzidos por cocaína parecem ser mediados pela ativação do receptor CB1 e inibidos pela ativação do receptor CB2. Considerando que ambos receptores são acoplados à proteína Gi/o, especulamos que estes receptores atuem em tipos distintos de neurônios, estando CB1 e CB2 localizados em neurônios GABAérgicos e dopaminérgicos, respectivamente. Drugs of abuse, such as cocaine, induce a reward effect and facilitate dopaminergic neurotransmission on the mesolimbic pathway. The endocannabinoid system, represented by cannabinoid receptors (CB1 e CB2), endocannabinoids and enzymes responsible for their synthesis and degradation, modulates the neurotransmission pathway and can be an important pharmacological target for the treatment of addiction. Herein, we tested the hypothesis that cannabinoid receptors are implicated on motor sensitization effects, conditioned place preference (CPP) and hippocampal neuronial activation induced by cocaine. Swiss male mice received AM251 (CB1 receptor antagonist) or JWH133 (CB2 receptor agonist) injections during acquisition or expression phases of the above mentioned behavioural tasks. AM251 inhibited sensitization, when administered during the acquisition (0,3, 1 and 3 mg/kg) or expression. (1 and 3 mg/kg) phases, and CPP, when administered during acquisition phase (10 mg/kg). JWH133 did not prevent the acute cocaine-effect. However, it attenuated sensitization when administered during acquisition (0,3 and 1 mg/kg) or expression (1 and 3 mg/kg) phases. Moreover, administration of JWH133 (10 mg/kg) prevented cocaine-induced CPP on both phases. JWH133 effects were reverted by AM630 (CB2 receptor antagonist) administration. The attenuation of the cocaine-induced contextual memory occurred along with a decrease on neuronal activation (c-Fos positive cells) on the hippocampus of animals treated with AM251 and JWH133. Therefore, behavioural and cellular effects induced by cocaine seem to be mediated by activation of CB1 receptor and inhibited by activation of CB2 receptor. As both receptors are Gi/o coupled receptors, we speculate that they act on different neuronal types, with CB1 e CB2 localized on GABAergic and dopaminergic neurons, respectively.

Although epidemiological studies have reported an association between hypercholesterolemia and mood disorders, there is a lack of data regarding depressive-like behavior in animal models of hypercholesterolemia. To address these questions, we assessed depressive-like behavior and hippocampal and cortical monoaminergic metabolism in three-month-old, low-density lipoprotein receptor knockout (LDLr(-/-)) and C57BL/6 wild-type mice. The LDLr(-/-) mice exhibited depressive-like behavior in the sucrose preference test, splash test, and tail suspension test. Increased monoamine oxidase (MAO) A and B activity was evidenced in the hippocampus of LDLr(-/-) mice. Furthermore, to address whether or not cholesterol modulates MAO activity, we exposed SH-SY5Y human neuroblastoma cells to human isolated low-density lipoprotein (LDL). Notably, LDL increased the activity of MAO-A and stimulated the reactive species generation in vitro. These findings indicate that depressive-like behavior in hypercholesterolemic mice is accompanied by alterations in the monoaminergic metabolism, providing new evidence about the association between hypercholesterolemia and depression.

Aims Epidemiological findings demonstrated that increased plasma cholesterol levels are frequently observed in depressive patients. In this regard, there is enhancing evidence that hypercholesterolemia is associated with impairment of brain function. Recently, we demonstrated that low-density lipoprotein receptor knockout (LDLr-/-) mice- a widely used rodent model of familial hypercholesterolemia - exhibited memory deficits and cortico-cerebral mitochondrial dysfunction. In this study, we aimed to assess the hypercholesterolemic mice in predictive tasks for depressive-like behavior. Methods Adult wild type C57BL/6 and LDLr -/- mice were evaluated in two tests for depressive like behavior, the splash test and forced swimming test. In addition, the activity of monoamine oxidase isoforms and the mRNA levels of hemeoxygenase-1 were assessed in the hippocampus and cerebral cortex of C57BL/6 and LDLr -/- mice. Finally, the blood-brain-barrier (BBB) permeability was investigated using the AQP-4 immunofluorescence staining in the mice hippocampus. Results The LDLr -/- mice showed a significant reduction in the grooming time in the splash test and increased immobility time in the forced swimming test, and both parameters were reversed by fluoxetine antidepressant treatment (10 mg/kg, 7 days, o.g.). Interestingly, the depressive like behavior of LDLr -/- mice was associated with increased activity of monoamine oxidase A, decreased hemeoxygenase-1 mRNA levels and increase of BBB permeability in the hippocampus. Conclusions Overall, these data provide new evidence that hypercholesterolemia could trigger brain alterations involved in depressive disorders.