SUMOylation effects on neural stem cells self-renewal, differentiation, and survival.

SUMO (small ubiquitin-like modifier) conjugation or SUMOylation, a post-translational modification, is a crucial regulator of protein function and cellular processes. In the context of neural stem cells (NSCs), SUMOylation has emerged as a key player, affecting their proliferation, differentiation, and survival. By modifying transcription factors, such as SOX1, SOX2, SOX3, SOX6, Bmi1, and Nanog, SUMOylation can either enhance or impair their transcriptional activity, thus impacting on NSCs self-renewal. Moreover, SUMOylation regulates neurogenesis and neuronal differentiation by modulating key proteins, such as Foxp1, Mecp2, MEF2A, and SOX10. SUMOylation is also crucial for the survival and proliferation of NSCs in both developing and adult brains. By regulating the activity of transcription factors, coactivators, and corepressors, SUMOylation acts as a molecular switch, inducing cofactor recruitment and function during development. Importantly, dysregulation of NSCs SUMOylation has been implicated in various disorders, including embryonic defects, ischemic cerebrovascular disease, glioma, and the harmful effects of benzophenone-3 exposure. Here we review the main findings on SUMOylation-mediated regulation of NSCs self-renewal, differentiation and survival. Better understanding NSCs SUMOylation mechanisms and its functional consequences might provide new strategies to promote neuronal differentiation that could contribute for the development of novel therapies targeting neurodegenerative diseases.

Neuroprotective potential of the Amazonian fruits Euterpe oleracea Mart. and Paullinia cupana Kunth

Abstract Acai (Euterpe oleracea Mart.) and guarana (Paullinia cupana Kunth) are native species from the Amazon Forest that in folk medicine are used to treat several diseases due to their anti-inflammatory and antioxidant properties. This review brings together findings from different studies on the potential neuroprotective effects of acai and guarana, highlighting the importance of the conservation and sustainable exploitation of the Amazon Forest. A bibliographic survey in the PubMed database retrieved indexed articles written in English that focused on the effects of acai and guarana in in vitro and in vivo models of neurodegenerative diseases. In general, treatment with either acai or guarana decreased neuroinflammation, increased antioxidant responses, ameliorated depression, and protected cells from neurotoxicity mediated by aggregated proteins. The results from these studies suggest that flavonoids, anthocyanins, and carotenoids found in both acai and guarana have therapeutic potential not only for neurodegenerative diseases, but also for depressive disorders. In addition, acai and guarana show beneficial effects in slowing down the physiological aging process. However, toxicity and efficacy studies are still needed to guide the formulation of herbal medicines from acai and guarana.

A nationwide study on sleep complaints and associated factors in older adults: ELSI-Brazil / Estudo nacional sobre queixas de sono e fatores associados em idosos: ELSI-Brasil / Estudio nacional sobre quejas de sueño y factores asociados en adultos mayores: ELSI-Brasil

Abstract: Sleep problems, such as difficulty falling asleep, staying asleep, early awakening with failure to continue sleep, and altered sleep-wake cycle, are common in the general population. This cross-sectional study with 6,929 older adults (≥ 60 years) aimed to estimate the prevalence of different types of sleep problems, their associated factors, and the population-attributable fraction of associated factors among older adults. The outcome variables consisted of self-reported sleep problems: insomnia (initial, intermediate, late, and any type of insomnia), poor sleep quality, and daytime sleepiness. The independent variables were sociodemographic and behavioral characteristics and health conditions. The prevalence proportions were initial insomnia (49.1%), intermediate insomnia (49.2%), late insomnia (45.9%), any type of insomnia (58.6%), poor sleep quality (15.6%), and daytime sleepiness (38.4%). Female sex, presence of two or more chronic diseases, not eating the recommended amount of fruits and vegetables, and regular and bad/very bad self-rated health were positively associated with the sleep problems investigated. Consuming alcohol once a month or more was inversely associated with initial insomnia. Population attributable fraction estimates ranged from 3% to 19% considering two or more chronic diseases, not eating the recommended amount of fruits and vegetables, and regular and bad/very bad self-rated health. High prevalence of self-reported sleep problems was evinced in older adults. These results can be useful to guide public health services in the creation of informational, evaluative, and screening strategies for sleep problems in older Brazilian adults.

Resumo: Problemas de sono, como dificuldade para adormecer, permanecer dormindo, despertar precoce com falha na continuidade do sono e alteração do ciclo vigília-sono, são comuns na população em geral. Este estudo transversal com 6.929 idosos (≥ 60 anos) buscou estimar a prevalência de diferentes tipos de problemas de sono, seus fatores associados e a fração atribuível populacional de fatores associados a problemas de sono nessa população. As variáveis de desfecho foram problemas de sono autorreferidos: insônia (inicial, intermediária, tardia e qualquer tipo de insônia), má qualidade do sono e sonolência diurna. As variáveis independentes incluíram características sociodemográficas, comportamentais e condições de saúde. As proporções de prevalência foram: insônia inicial (49,1%), insônia intermediária (49,2%), insônia tardia (45,9%), qualquer tipo de insônia (58,6%), má qualidade do sono (15,6%) e sonolência diurna (38,4%). Sexo feminino, presença de duas ou mais doenças crônicas, não consumir a quantidade recomendada de frutas e hortaliças e autoavaliação da saúde como regular e ruim/muito ruim mostraram associação positiva aos problemas de sono investigados. Consumo de álcool uma vez por mês ou mais associou-se inversamente à insônia inicial. As estimativas da fração atribuível populacional variaram de 3% a 19% considerando duas ou mais doenças crônicas, consumo insuficiente de frutas e vegetais e saúde autorrelatada regular/ruim/muito ruim. Evidenciou-se alta prevalência de problemas de sono autorreferidos em idosos. Esses resultados podem orientar os serviços públicos de saúde na criação de estratégias informativas, avaliativas e de rastreamento de problemas de sono em idosos brasileiros.

Resumen: Problemas del sueño, como la dificultad para conciliar el sueño, permanecer dormido, despertarse temprano sin poder seguir durmiendo y cambios en el ciclo de sueño y vigilia, son comunes en la población en general. Este estudio transversal con 6.929 personas mayores (≥ 60 años) buscó estimar la prevalencia de diferentes tipos de problemas de sueño, sus factores asociados y la fracción atribuible a la población de factores asociados con problemas de sueño en esta población. Las variables de desenlace fueron problemas de sueño autoinformados: insomnio (inicial, intermedio, tardío y cualquier tipo de insomnio), mala calidad del sueño y somnolencia diurna. Las variables independientes incluyeron características sociodemográficos y conductuales y condiciones de salud. Estas fueron las proporciones de prevalencia: insomnio inicial (49,1%), insomnio intermedio (49,2%), insomnio tardío (45,9%), cualquier tipo de insomnio (58,6%), mala calidad del sueño (15,6%) y somnolencia diurna (38,4%). El sexo femenino, la presencia de dos o más enfermedades crónicas, no consumir la cantidad recomendada de frutas y hortalizas y la autoevaluación de la salud como regular y mala/muy mala mostraron una asociación positiva con los problemas de sueño investigados. El consumo de alcohol una vez al mes o más se asoció inversamente con el insomnio inicial. Las estimaciones de la fracción atribuible de la población oscilaron entre el 3% y el 19% considerando dos o más enfermedades crónicas, un consumo insuficiente de frutas y verduras y una salud autoinformada regular/mala/muy mala. Se evidenció una alta prevalencia de problemas de sueño autoinformados en las personas mayores. Estos resultados pueden orientar los servicios públicos de salud en la creación de estrategias informativas, evaluativas y de seguimiento de los problemas de sueño en las personas mayores brasileñas.

Guanosine modulates SUMO2/3-ylation in neurons and astrocytes via adenosine receptors.

SUMOylation is a post-translational modification (PTM) whereby members of the Small Ubiquitin-like MOdifier (SUMO) family of proteins are conjugated to lysine residues in target proteins. SUMOylation has been implicated in a wide range of physiological and pathological processes, and much attention has been given to its role in neurodegenerative conditions. Due to its reported role in neuroprotection, pharmacological modulation of SUMOylation represents an attractive potential therapeutic strategy in a number of different brain disorders. However, very few compounds that target the SUMOylation pathway have been identified. Guanosine is an endogenous nucleoside with important neuromodulatory and neuroprotective effects. Experimental evidence has shown that guanosine can modulate different intracellular pathways, including PTMs. In the present study we examined whether guanosine alters global protein SUMOylation. Primary cortical neurons and astrocytes were treated with guanosine at 1, 10, 100, 300, or 500 µM at four time points, 1, 6, 24, or 48 h. We show that guanosine increases global SUMO2/3-ylation in neurons and astrocytes at 1 h at concentrations above 10 µM. The molecular mechanisms involved in this effect were evaluated in neurons. The guanosine-induced increase in global SUMO2/3-ylation was still observed in the presence of dipyridamole, which prevents guanosine internalization, demonstrating an extracellular guanosine-induced effect. Furthermore, the A1 adenosine receptor antagonist DPCPX abolished the guanosine-induced increase in SUMO2/3-ylation. The A2A adenosine receptor antagonist ZM241385 increased SUMOylation per se, but did not alter guanosine-induced SUMOylation, suggesting that guanosine may modulate SUMO2/3-ylation through an A1-A2A receptor interaction. Taken together, this is the first report to show guanosine as a SUMO2/3-ylation enhancer in astrocytes and neurons.

Enhanced SUMOylation and SENP-1 protein levels following oxygen and glucose deprivation in neurones.

Here, we show that oxygen and glucose deprivation (OGD) causes increased small ubiquitin-like modifier (SUMO)-1 and SUMO-2/3 conjugation to substrate proteins in cultured hippocampal neurones. Surprisingly, the SUMO protease SENP-1, which removes SUMO from conjugated proteins, was also increased by OGD, suggesting that the neuronal response to OGD involves a complex interplay between SUMOylation and deSUMOylation. Importantly, decreasing global SUMOylation in cultured hippocampal neurones by overexpression of the catalytic domain of SENP-1 increased neuronal vulnerability to OGD-induced cell death. Taken together, these results suggest a neuroprotective role for neuronal SUMOylation after OGD.

Oxygen/glucose deprivation induces a reduction in synaptic AMPA receptors on hippocampal CA3 neurons mediated by mGluR1 and adenosine A3 receptors.

Hippocampal CA1 pyramidal neurons are highly sensitive to ischemic damage, whereas neighboring CA3 pyramidal neurons are less susceptible. It is proposed that switching of AMPA receptor (AMPAR) subunits on CA1 neurons during an in vitro model of ischemia, oxygen/glucose deprivation (OGD), leads to an enhanced permeability of AMPARs to Ca(2+), resulting in delayed cell death. However, it is unclear whether the same mechanisms exist in CA3 neurons and whether this underlies the differential sensitivity to ischemia. Here, we investigated the consequences of OGD for AMPAR function in CA3 neurons using electrophysiological recordings in rat hippocampal slices. Following a 15 min OGD protocol, a substantial depression of AMPAR-mediated synaptic transmission was observed at CA3 associational/commissural and mossy fiber synapses but not CA1 Schaffer collateral synapses. The depression of synaptic transmission following OGD was prevented by metabotropic glutamate receptor 1 (mGluR1) or A(3) receptor antagonists, indicating a role for both glutamate and adenosine release. Inhibition of PLC, PKC, or chelation of intracellular Ca(2+) also prevented the depression of synaptic transmission. Inclusion of peptides to interrupt the interaction between GluA2 and PICK1 or dynamin and amphiphysin prevented the depression of transmission, suggesting a dynamin and PICK1-dependent internalization of AMPARs after OGD. We also show that a reduction in surface and total AMPAR protein levels after OGD was prevented by mGluR1 or A(3) receptor antagonists, indicating that AMPARs are degraded following internalization. Thus, we describe a novel mechanism for the removal of AMPARs in CA3 pyramidal neurons following OGD that has the potential to reduce excitotoxicity and promote neuroprotection.

Lithium and valproate protect hippocampal slices against ATP-induced cell death.

Lithium and valproate (VPA) are the most commonly prescribed mood-stabilizing drugs. Recently, several studies have reported their neuroprotective properties in several models of neural toxicity and, in some pathological conditions, large amounts of intracellular ATP can be released from damaged cells. In the present study, we investigate the potential neuroprotective effect of lithium and VPA against ATP-induced cell death in hippocampal slices of adult rats. Acute (in vitro) and chronic (in vivo) treatment at therapeutic doses with lithium or VPA significantly prevent the ATP-induced cell death. Lithium and VPA also exerted a synergic effect in the prevention of ATP-induced cell death. Moreover, hippocampal slices prepared from rats chronically treated with lithium or VPA presented a significant reduction in cell death in the presence of cytotoxic extracellular ATP. Although further investigations are necessary, our results show the neuroprotective effect of lithium and VPA against neuronal death induced by extracellular ATP, probably through a different pathway, and suggest novel uses of these drugs in neurogenerative diseases.

The effects of estradiol on estrogen receptor and glutamate transporter expression in organotypic hippocampal cultures exposed to oxygen--glucose deprivation.

The molecular basis of estrogen-mediated neuroprotection against brain ischemia remains unclear. In the present study, we investigated changes in expression of estrogen receptors (ERs) alpha and beta and excitatory amino acid transporters (EAAT) 1 and 2 in rat organotypic hippocampal slice cultures treated with estradiol and subsequently exposed to oxygen--glucose deprivation (OGD). Pretreatment with 17beta-estradiol (10 nM) for 7 days protected the CA1 area of hippocampus against OGD (60 min), reducing cellular injury by 46% compared to the vehicle control group. Levels of ERalpha protein were significantly reduced by 20% after OGD in both vehicle- and estradiol-treated cultures, whereas ERbeta was significantly up-regulated by 25% in the estradiol-treated cultures. In contrast, EAAT1 and EAAT2 levels were unchanged in response to estradiol treatment in this model of OGD. These findings suggest that estrogen-induced neuroprotection against ischemia might involve regulation of ERbeta and, consequently, of the genes influenced by this receptor.

Neuroprotection and protein damage prevention by estradiol replacement in rat hippocampal slices exposed to oxygen-glucose deprivation.

Here we investigated the effects of estradiol replacement in ovariectomized female rats using hippocampal slices exposed to oxygen-glucose deprivation (OGD). OGD induced lactate dehydrogenase (LDH) release to the incubation medium, what was assumed as a parameter of cellular death. In the estradiol-treated group the LDH release was markedly decreased by 23% as compared to the vehicle-treated group. In attempt to study a possible mechanism by which estradiol acts, we investigated some parameters of oxidative stress. In both vehicle-treated and estradiol-treated groups, OGD significantly increased the free radical production by 34% and 16%, respectively, although no significant differences on total antioxidant capacity were observed. Interestingly, estradiol replacement prevented the significant reduction in tryptophan and tyrosine contents caused by OGD observed in vehicle-treated animals. Our results show that estradiol replacement in ovariectomized female rats decreases cellular susceptibility to an ischemic-like injury and suggest a role for the hormone on protein damage prevention.

Cellular death in hippocampus in response to PI3K pathway inhibition and oxygen and glucose deprivation.

We investigated the importance of the phosphoinositide3-kinase (PI3K) pathway in CA1 and dentate gyrus (DG) areas of hippocampus by exposing organotypic cultures to LY294002, a PI3K inhibitor, or to oxygen and glucose deprivation (OGD) for up to 21 hours. LY294002 induced increased propidium iodide (PI) uptake and caspase 3/7 activity in both regions, with a faster onset in DG. In contrast, cultures exposed to 60 min of OGD showed a PI uptake only in the CA1 area, beginning 13 h after the insult and increasing until 21 h. We did not observe any significant changes in AKT phosphorylation and immunocontent in CA1 or DG areas of organotypic cultures exposed to OGD, suggesting that the phosphorylation of this protein at Ser-473 is unrelated to the cellular damage induced by ischemia. Our results suggest that the inhibition of the PI3K pathway does not mimic the cell death profile observed with an ischemic model.

Hypoxic preconditioning in neonatal rat brain involves regulation of excitatory amino acid transporter 2 and estrogen receptor alpha.

Exposure of the brain to a sublethal insult can protect against a subsequent brain injury. Hypoxic preconditioning induces tolerance to hypoxic--ischemic injury in neonatal rat brain and is associated with changes in gene and protein expression. To study the involvement of excitatory amino acid transporters (EAAT1 and EAAT2) and estrogen receptors (ERalpha and ERbeta) in neonatal hypoxia--induced ischemic tolerance, we examined changes in expression of these proteins in the cortex, hippocampus and striatum of newborn rats at different time points after exposure to sublethal hypoxia (8% O(2), 3h). Preconditioning with hypoxia 24h before hypoxia-ischemia afforded marked brain protection compared with littermate control animals as determined by morphological assessment. Immunoblot analysis showed that EAAT2 and ERalpha were significantly increased by 55% and 49%, respectively, in cortex at 24h after hypoxic-preconditioning. Surprisingly, at the same time point, a significant decrease of EAAT2 by 48% in striatum was observed. In contrast, hypoxic preconditioning had no effect on the levels of EAAT1 and ERbeta in any of the brain regions studied at any of the time points analyzed. The similar pattern of changes in EAAT2 and ERalpha levels suggests that ERalpha might interact with EAAT2 in producing preconditioning. The endogenous molecular mechanisms modulated by hypoxia preconditioning may contribute to the development of hypoxia-induced ischemic tolerance, and may provide novel therapeutic targets for the treatment of cerebral ischemia.

Estradiol protects against oxygen and glucose deprivation in rat hippocampal organotypic cultures and activates Akt and inactivates GSK-3beta.

Here we investigated the neuroprotective effect of 17beta-estradiol in an in vitro model of ischemia. We used organotypic hippocampal slice cultures, acute or chronically treated with 17beta-estradiol (10 nM), and exposed to oxygen and glucose deprivation (OGD). Cellular death was quantified by measuring uptake of propidium iodide (PI), a marker of dead cells. In OGD exposed cultures, treated only with vehicle, about 70% of the CA1 area of hippocampus was labeled with PI, indicating a great percentage of cellular death. When cultures were treated with 17beta-estradiol (acute or chronically), this cellular death was reduced to 15%. This effect was prevented by LY294002 but was not by PD98059. Immunoblotting revealed that both, chronic and acute, treatments with 17beta-estradiol induced the phosphorylation/activation of Akt and the phosphorylation/inactivation of GSK-3beta. Our results show a clear neuroprotective effect of 17beta-estradiol and suggest that this effect could involve PI3-K pathway.