Sex-dependent effect on mitochondrial and oxidative stress parameters in the hypothalamus induced by prepubertal stress and access to high fat diet

Objective: Some factors related to lifestyle, including stress and high-fat diet (HFD) consumption, are associated with higher prevalence of obesity. These factors can lead to an imbalance between ROS production and antioxidant defenses and to mitochondrial dysfunctions, which, in turn, could cause metabolic impairments, favoring the development of obesity. However, little is known about the interplay between these factors, particularly at early ages, and whether long-term sex-specific changes may occur. Here, we evaluated whether social isolation during the prepubertal period only, associated or not with chronic HFD, can exert long-term effects on oxidative status parameters and on mitochondrial function in the whole hypothalamus, in a sex-specific manner. Methods: Wistar male and female rats were divided into two groups (receiving standard chow or standard chow + HFD), that were subdivided into exposed or not to social isolation during the prepubertal period. Oxidative status parameters, and mitochondrial function were evaluated in the hypothalamus in the adult age. Results: Regarding antioxidant enzymes activities, HFD decreased GPx activity in the hypothalamus, while increasing SOD activity in females. Females also presented increased total thiols; however, non-protein thiols were lower. Main effects of stress and HFD were observed in TBARS levels in males, with both factors decreasing this parameter. Additionally, HFD increased complex IV activity, and decreased mitochondrial mass in females. Complex I-III activity was higher in males compared to females. Conclusion: Stress during the prepubertal period and chronic consumption of HFD had persistent sex-specific effects on oxidative status, as well as on its consequences for the cell and for mitochondrial function. HFD had more detrimental effects on females, inducing oxidative imbalance, which resulted in damage to the mitochondria. This HFD-induced imbalance may be related to the development of obesity

Cognitive development and brain gray matter susceptibility to prenatal adversities : moderation by the prefrontal cortex brain-derived neurotrophic factor gene co-expression network

Background: Previous studies focused on the relationship between prenatal conditions and neurodevelopmental outcomes later in life, but few have explored the interplay between gene co-expression networks and prenatal adversity conditions on cognitive development trajectories and gray matter density. Methods: We analyzed the moderation effects of an expression polygenic score (ePRS) for the Brain-derived Neurotrophic Factor gene network (BDNF ePRS) on the association between prenatal adversity and child cognitive development. A score based on genes co-expressed with the prefrontal cortex (PFC) BDNF was created, using the effect size of the association between the individual single nucleotide polymorphisms (SNP) and the BDNF expression in the PFC. Cognitive development trajectories of 157 young children from the Maternal Adversity, Vulnerability and Neurodevelopment (MAVAN) cohort were assessed longitudinally in 4-time points (6, 12, 18, and 36 months) using the Bayley-II mental scales. Results: Linear mixed-effects modeling indicated that BDNF ePRS moderates the effects of prenatal adversity on cognitive growth. In children with high BDNF ePRS, higher prenatal adversity was associated with slower cognitive development in comparison with those exposed to lower prenatal adversity. Parallel-Independent Component Analysis (pICA) suggested that associations of expression-based SNPs and gray matter density significantly differed between low and high prenatal adversity groups. The brain IC included areas involved in visual association processes (Brodmann area 19 and 18), reallocation of attention, and integration of information across the supramodal cortex (Brodmann area 10). Conclusion: Cognitive development trajectories and brain gray matter seem to be influenced by the interplay of prenatal environmental conditions and the expression of an important BDNF gene network that guides the growth and plasticity of neurons and synapses

Leptin receptor co-expression gene network moderates the effect of early life adversity on eating behavior in children

Leptin influences eating behavior. Exposure to early adversity is associated with eating behaviour disorders and metabolic syndrome, but the role of the leptin receptor on this relationship is poorly explored. We investigated whether individual differences in brain region specific leptin receptor (LepR) gene networks could moderate the effects of early adversity on eating behavior and metabolism. We created an expression-based polygenic risk score (ePRS) reflecting variations in the function of LepR gene network in prefrontal cortex and hypothalamus to investigate the interactions between a cumulative index of postnatal adversity on eating behavior in two independent birth cohorts (MAVAN and GUSTO). To explore whether variations in the prefrontal cortex or hypothalamic genetic scores could be associated with metabolic measurements, we also assessed the relationship between LepR-ePRS and fasting blood glucose and leptin levels in a third independent cohort (ALSPAC). We identified significant interaction effects between postnatal adversity and prefrontal-based LepR-ePRS on the Child Eating Behavior Questionnaire scores. In MAVAN, we observed a significant interaction effect on food enjoyment at 48 months (β = 61.58, p = 0.015) and 72 months (β = 97.78, p = 0.001); food responsiveness at 48 months (β = 83.79, p = 0.009) satiety at 48 months (β = −43.63, p = 0.047). Similar results were observed in the GUSTO cohort, with a significant interaction effect on food enjoyment (β = 30.48, p = 0.006) food fussiness score (β = −24.07, p = 0.02) and satiety score at 60 months (β = −17.00, p = 0.037). No effects were found when focusing on the hypothalamus-based LepR-ePRS on eating behavior in MAVAN and GUSTO cohorts, and there was no effect of hypothalamus and prefrontal cortex based ePRSs on metabolic measures in ALSPAC. Our study indicated that exposure to postnatal adversity interacts with prefrontal cortex LepR-ePRS to moderate eating behavior, suggesting a neurobiological mechanism associated with the development of eating behavior problems in response to early adversity. The knowledge of these mechanisms may guide the understanding of eating patterns associated with risk for obesity in response to fluctuations in stress exposure early in life

Amygdala 5-HTT gene network moderates the effects of postnatal adversity on attention problems : anatomo-functional correlation and epigenetic changes

Variations in serotoninergic signaling have been related to behavioral outcomes. Alterations in the genome, such as DNA methylation and histone modifications, are affected by serotonin neurotransmission. The amygdala is an important brain region involved in emotional responses and impulsivity, which receives serotoninergic input. In addition, studies suggest that the serotonin transporter gene network may interact with the environment and influence the risk for psychiatric disorders. We propose to investigate whether/how interactions between the exposure to early life adversity and serotonin transporter gene network in the amygdala associate with behavioral disorders. We constructed a co-expression-based polygenic risk score (ePRS) reflecting variations in the function of the serotonin transporter gene network in the amygdala and investigated its interaction with postnatal adversity on attention problems in two independent cohorts from Canada and Singapore. We also described how interactions between ePRS-5-HTT and postnatal adversity exposure predict brain gray matter density and variation in DNA methylation across the genome. We observed that the expression-based polygenic risk score, reflecting the function of the amygdala 5-HTT gene network, interacts with postnatal adversity, to predict attention and hyperactivity problems across both cohorts. Also, both postnatal adversity score and amygdala ePRS-5-HTT score, as well as their interaction, were observed to be associated with variation in DNA methylation across the genome. Variations in gray matter density in brain regions linked to attentional processes were also correlated to our ePRS score. These results confirm that the amygdala 5-HTT gene network is strongly associated with ADHD-related behaviors, brain cortical density, and epigenetic changes in the context of adversity in young children

Isolamento social precoce e consumo de uma dieta hiperlipídica : implicações no comportamento do tipo depressivo e em aspectos cognitivos em ratos adultos

Intervenções ambientais, como a exposição precoce a estressores ou a dietas ricas em calorias, podem alterar a trajetória da maturação neural e influenciar na susceptibilidade a certas patologias a longo-prazo. Neste contexto, o objetivo do presente estudo foi investigar os efeitos de uma exposição ao isolamento social durante o período pré-pubere associado ou não ao consumo precoce e crônico de uma dieta hiperlipídica (HFD) sobre aspectos cognitivos e emocionais, e possíveis mecanismos neuroquímicos associados a essas alterações, no hipocampo, no córtex pré-frontal e no núcleo accumbens de ratos machos na idade adulta. Os resultados mostraram que os dois fatores, estresse e dieta (separadamente), induziram um comportamento do tipo depressivo nos animais na idade adulta. Além disso, os animais isolados apresentaram um déficit cognitivo associado à memória de curta-duração e de trabalho, enquanto que animais com acesso à HFD demonstraram prejuízo somente na memória de curta-duração. Curiosamente, a interação entre os fatores (estresse e dieta) causou uma reversão dos déficits na memória de curta-duração. Em relação às avaliações do comportamento alimentar hedônico, observamos que o grupo com consumo crônico de HFD apresentou uma menor motivação para obter diferentes tipos de alimentos palatáveis doces. Essa redução motivacional não parece ser associada a uma menor palatabilidade e/ou a uma maior saciedade induzida pela HFD. Em relação aos marcadores de plasticidade analisados no córtex pré-frontal, observamos interações entre os fatores estresse e dieta na atividade da enzima Na+K+-ATPase, nos níveis de BNDF e no imunoconteúdo das proteínas AKT e MAPK/ERK, sendo que os fatores quando aplicados isolados diminuem os níveis dos parâmetros analisados, porém quando associados, os níveis retornam ou aumentam em relação aos valores do grupo controle. O hipocampo foi a estrutura mais afetada pelas intervenções ambientais neste trabalho. Observamos que tanto o estresse, como a dieta hiperlipídica (separadamente) causaram uma redução da plasticidade sináptica hipocampal, por meio de diferentes mecanismos: o acesso crônico à HFD afeta proteínas relacionadas ao funcionamento das sinapses, enquanto o isolamento social parece afetar mais particularmente a via de sinalização do BDNF. Com relação aos achados neuroquímicos no núcleo accumbens, observamos uma redução dos receptores dopaminérgico-1 (D1) e canabinóide-1 (CB1) com o consumo de HFD, enquanto que os animais isolados na pré-puberdade apresentaram uma redução do metabolismo dopaminérgico nesta estrutura. Em suma, esta tese demonstra que tanto o isolamento social e como o consumo contínuo de HFD causam comportamento do tipo depressivo e outras alterações comportamentais, e reduzem marcadores de plasticidade importantes no córtex prefrontal e hipocampo. O acesso à HFD também causa uma menor motivação para o comportamento alimentar hedônico. Assim, tais eventos precoces podem afetar a plasticidade neural levando a importantes alterações comportamentais, e assim predispor a diferentes patologias durante a vida.Environmental interventions, such as early exposure to stressors or high calorie-diets, can alter the trajectory of neural maturation and influence the susceptibility to some pathologies throughout life. In this context, the aim of the present study was to investigate the effects of exposure to social isolation, during the pre-pubertal period, associated or not with early and chronic consumption of a hyperlipid diet (HFD) on cognitive and emotional aspects, and possible mechanisms associated with these changes, in the hippocampus, the prefrontal cortex and the nucleus accumbens of male rats in adulthood. The results showed that both pre-pubertal social isolation and chronic access to a hyperlipidic diet induced a depressive-like behavior in animals during adulthood. In addition, isolated animals had a cognitive deficit associated with short-term and working memory, whereas animals with access to HFD demonstrated impairment only in short-term memory. Interestingly, the interaction between the factors (stress and diet) caused a reversal in relation to short-term memory, remaining similar to the control group. Regarding the evaluations of the hedonic eating behavior, we observed that HFD group presented a lower motivation to obtain different sweet palatable foods. This impaired motivation does not appear to be associated with less palatability and/or satiety induced by the high-fat diet. Concerning plasticity markers in the prefrontal cortex, we observed interactions between stress and diet on Na+ K+-ATPase activity, BNDF levels and AKT and MAPK/ERK immunocontents. These interactions follow a similar profile: when applied alone, the levels of the analyzed parameters decrease, but when associated, the levels return or increase in relation to the values of the control group. The hippocampus was the structure most affected by the environmental interventions. Both stress and HFD caused a reduction of hippocampal synaptic plasticity through different mechanisms: chronic access to HFD affects proteins related to synaptic function, while social isolation affects the BDNF signaling pathway more significantly. Regarding the neurochemical findings in the nucleus accumbens, we observed a reduction in dopaminergic-1 (D1) and cannabinoid-1 (CB1) receptors with chronic HFD intake, whereas isolated animals had a reduction of dopaminergic metabolism in this same structure. In summary, this thesis shows that both social isolation and chronic consumption of HFD lead to depressive-like behavior and to other behavioral changes; they reduce plasticity markers in prefrontal cortex and hippocampus. HFD access also induced a lower motivation for hedonic feeding. Therefore, these early interventions may affect neural plasticity, leading to important behavioral changes, and thus, predispose to different pathologies later in life

Estresse no período pré-pubere e exposição crônica a dietas hiperpalatáveis : avaliação do comportamento do tipo ansioso e de alterações bioquímicas em ratos machos adultos

A pré-puberdade é um período crítico para a maturação final dos circuitos neuronais que controlam a homeostase energética e as respostas ao estresse. Exposição a estressores e a dietas hiperpalatáveis neste período de desenvolvimento podem modificar o processo de maturação e causar mudanças comportamentais e neuroquímicas na idade adulta. Desta forma, o objetivo deste estudo é verificar os efeitos da exposição ao estresse por isolamento social no período pré-pubere e o acesso crônico a dietas hiperpalatáveis sobre comportamento do tipo ansioso e alterações metabólicas e neuroquímicas, em ratos machos adultos. Os animais que foram isolados apresentaram um comportamento do tipo ansiolítico no Labirinto em Cruz Elevado, e o mesmo foi observado nos animais isolados com acesso a dieta hiperlipídica. Já os animais estressados com acesso a dieta rica em carboidratos apresentaram comportamento oposto, ou seja, do tipo ansiogênico. Foram observadas mudanças metabólicas principalmente nos animais que receberam dieta rica em gordura (aumento do peso corporal, gordura abdominal, assim como aumento da glicose plasmática e da atividade da colinestarese plasmática) e a maioria desses efeitos foram aumentados com a exposição ao estresse. A dieta hiperlipídica associado ao estresse também afetou o perfil lipídico aumentando LDL-colesterol nesses animais. Além disso, exposição ao estresse levou a um desequilíbrio oxidativo no fígado, com aumento da produção de espécies reativas, assim como aumento da atividade de enzimas antioxidantes (Superóxido dismutase e Catalase), e esses efeitos foram acentuados com o acesso à dieta hiperlipídica (que também causou uma grave redução na atividade da Glutationa peroxidase). No hipocampo, o estresse levou a uma diminuição da atividade das enzimas antioxidantes, do conteúdo de tióis totais e da atividade dos complexos II e IV da cadeira respiratória mitocondrial. A dieta hiperlipídica quando associada ao estresse reverteu esses efeitos. A partir dos resultados encontrados, concluiu-se que o período pré-pubere representa um período crítico para intervenções durante o desenvolvimento, e o estresse nesse período leva a alterações comportamentais, metabólicas e neuroquímicas na idade adulta, diminuindo o comportamento do tipo ansioso e aumentando a suscetibilidade ao estresse oxidativo tanto no fígado como hipocampo. Esse desfecho, porém dependem do tipo de dieta a que o animal tem acesso.The pre-puberty period is critical for the final maturation of neural circuits that control energy homeostasis and stress responses. Exposure to stressors and palatable diets in this period of development may alter the maturation process and cause behavioral and neurochemical changes in adulthood. Thus, the objective of this study is to investigate the effects of exposure to stress by social isolation in the prepubertal period and of chronic access to palatable diets on anxious-like behavior, and on metabolic and neurochemical changes in liver and hippocampus of adult male rats. The animals that were isolated showed an anxiolytic-like behavior in the Plus Maze test, and the same was observed in isolated animals with access to high-fat diet. The stressed animals with access to high-carbohydrate diet showed opposite behavior; in other words, they presented anxiogenic-like behavior. Metabolic changes were observed mainly in animals fed high-fat diet (increased body weight, abdominal fat, as well as increased plasma glucose and plasma cholinesterase activity), and most of these effects were further increased by exposure to stress. The high-fat diet associated with stress also affected the lipid profile by increasing LDL-cholesterol in these animals. Furthermore, exposure to stress led to an oxidative imbalance in the liver, by increasing production of reactive species, and the activity of antioxidant enzymes (Catalase and Superoxide dismutase), and these effects were accentuated with access to high-fat diet (which also caused a severe reduction in Glutathione peroxidase activity). In the hippocampus, stress led to decrease in antioxidant enzymes activities, total thiol content and activity of complexes II and IV of the mitochondrial respiratory chair. However, high-fat diet, when associated with stress, reversed these effects. Taken together, we concluded that the prepubertal period is a critical period for interventions during development, and stress during this period leads to behavioral, metabolic and neurochemical changes in adulthood, decreasing anxious-like behavior and increasing the susceptibility to oxidative stress in liver and hippocampus, and these effects will differ depending on the type of diet to which the animal has access

Early life interventions: evaluation of oxidative parameters in distinct cns structures in adult female rats

Early life events lead to a large number of behavioral and biochemical alterations in adulthood. The aim of this study is to verify wheter the release of gonadal hormones during puberty affects parameters of oxidative stress observed in adulthood in cerebral cortex, striatum and hypothalamus of female rats subjected to neonatal handling. Rats were exposed or not to neonatal handling (10min/day, first 10 days of life). Between 21-28 post-natal days, females from each litter were divided into the following groups: ovariectomy, sham, and intact (no surgery). When adults, parameters of oxidative stress were analyzed. The groups subjected to surgery (ovx and sham), showed increased production of free radicals by the method of oxidation of dichlorodihydrofluorescein (DCFH) in cerebral cortex and striatum. Decreased catalase activity was observed in the cerebral cortex, in the same groups. No effects of neonatal manipulation were observed in these structures. We conclude that the period after weaning constitute a critical window for stressful interventions during development, leading to alterations in parameters of oxidative stress in adulthood, and these effects are not influenced by estradiol and neonatal handling