Two-Month Voluntary Ethanol Consumption Promotes Mild Neuroinflammation in the Cerebellum but Not in the Prefrontal Cortex, Hippocampus, or Striatum of Mice.

Chronic ethanol exposure often triggers neuroinflammation in the brain's reward system, potentially promoting the drive for ethanol consumption. A main marker of neuroinflammation is the microglia-derived monocyte chemoattractant protein 1 (MCP1) in animal models of alcohol use disorder in which ethanol is forcefully given. However, there are conflicting findings on whether MCP1 is elevated when ethanol is taken voluntarily, which challenges its key role in promoting motivation for ethanol consumption. Here, we studied MCP1 mRNA levels in areas implicated in consumption motivation-specifically, the prefrontal cortex, hippocampus, and striatum-as well as in the cerebellum, a brain area highly sensitive to ethanol, of C57BL/6 mice subjected to intermittent and voluntary ethanol consumption for two months. We found a significant increase in MCP1 mRNA levels in the cerebellum of mice that consumed ethanol compared to controls, whereas no significant changes were observed in the prefrontal cortex, hippocampus, or striatum or in microglia isolated from the hippocampus and striatum. To further characterize cerebellar neuroinflammation, we measured the expression changes in other proinflammatory markers and chemokines, revealing a significant increase in the proinflammatory microRNA miR-155. Notably, other classical proinflammatory markers, such as TNFα, IL6, and IL-1ß, remained unaltered, suggesting mild neuroinflammation. These results suggest that the onset of neuroinflammation in motivation-related areas is not required for high voluntary consumption in C57BL/6 mice. In addition, cerebellar susceptibility to neuroinflammation may be a trigger to the cerebellar degeneration that occurs after chronic ethanol consumption in humans.

Chronic Voluntary Morphine Intake Is Associated with Changes in Brain Structures Involved in Drug Dependence in a Rat Model of Polydrug Use.

Chronic opioid intake leads to several brain changes involved in the development of dependence, whereby an early hedonistic effect (liking) extends to the need to self-administer the drug (wanting), the latter being mostly a prefrontal-striatal function. The development of animal models for voluntary oral opioid intake represents an important tool for identifying the cellular and molecular alterations induced by chronic opioid use. Studies mainly in humans have shown that polydrug use and drug dependence are shared across various substances. We hypothesize that an animal bred for its alcohol preference would develop opioid dependence and further that this would be associated with the overt cortical abnormalities clinically described for opioid addicts. We show that Wistar-derived outbred UChB rats selected for their high alcohol preference additionally develop: (i) a preference for oral ingestion of morphine over water, resulting in morphine intake of 15 mg/kg/day; (ii) marked opioid dependence, as evidenced by the generation of strong withdrawal signs upon naloxone administration; (iii) prefrontal cortex alterations known to be associated with the loss of control over drug intake, namely, demyelination, axonal degeneration, and a reduction in glutamate transporter GLT-1 levels; and (iv) glial striatal neuroinflammation and brain oxidative stress, as previously reported for chronic alcohol and chronic nicotine use. These findings underline the relevance of polydrug animal models and their potential in the study of the wide spectrum of brain alterations induced by chronic morphine intake. This study should be valuable for future evaluations of therapeutic approaches for this devastating condition.

Amelioration of morphine withdrawal syndrome by systemic and intranasal administration of mesenchymal stem cell-derived secretome in preclinical models of morphine dependence.

BACKGROUND: Morphine is an opiate commonly used in the treatment of moderate to severe pain. However, prolonged administration can lead to physical dependence and strong withdrawal symptoms upon cessation of morphine use. These symptoms can include anxiety, irritability, increased heart rate, and muscle cramps, which strongly promote morphine use relapse. The morphine-induced increases in neuroinflammation, brain oxidative stress, and alteration of glutamate levels in the hippocampus and nucleus accumbens have been associated with morphine dependence and a higher severity of withdrawal symptoms. Due to its rich content in potent anti-inflammatory and antioxidant factors, secretome derived from human mesenchymal stem cells (hMSCs) is proposed as a preclinical therapeutic tool for the treatment of this complex neurological condition associated with neuroinflammation and brain oxidative stress. METHODS: Two animal models of morphine dependence were used to evaluate the therapeutic efficacy of hMSC-derived secretome in reducing morphine withdrawal signs. In the first model, rats were implanted subcutaneously with mini-pumps which released morphine at a concentration of 10 mg/kg/day for seven days. Three days after pump implantation, animals were treated with a simultaneous intravenous and intranasal administration of hMSC-derived secretome or vehicle, and withdrawal signs were precipitated on day seven by i.p. naloxone administration. In this model, brain alterations associated with withdrawal were also analyzed before withdrawal precipitation. In the second animal model, rats voluntarily consuming morphine for three weeks were intravenously and intranasally treated with hMSC-derived secretome or vehicle, and withdrawal signs were induced by morphine deprivation. RESULTS: In both animal models secretome administration induced a significant reduction of withdrawal signs, as shown by a reduction in a combined withdrawal score. Secretome administration also promoted a reduction in morphine-induced neuroinflammation in the hippocampus and nucleus accumbens, while no changes were observed in extracellular glutamate levels in the nucleus accumbens. CONCLUSION: Data presented from two animal models of morphine dependence suggest that administration of secretome derived from hMSCs reduces the development of opioid withdrawal signs, which correlates with a reduction in neuroinflammation in the hippocampus and nucleus accumbens.

Methadone directly impairs central nervous system cells in vitro.

Methadone is a synthetic long-acting opioid that is increasingly used in the replacement therapy of opioid-addicted patients, including pregnant women. However, methadone therapy in this population poses challenges, as it induces cognitive and behavioral impairments in infants exposed to this opioid during prenatal development. In animal models, prenatal methadone exposure results in detrimental consequences to the central nervous system, such as: (i) increased neuronal apoptosis; (ii) disruption of oligodendrocyte maturation and increased apoptosis and (iii) increased microglia and astrocyte activation. However, it remains unclear whether these deleterious effects result from a direct effect of methadone on brain cells. Therefore, our goal was to uncover the impact of methadone on single brain cell types in vitro. Primary cultures of rat neurons, oligodendrocytes, microglia, and astrocytes were treated for three days with 10 µM methadone to emulate a chronic administration. Apoptotic neurons were identified by cleaved caspase-3 detection, and synaptic density was assessed by the juxtaposition of presynaptic and postsynaptic markers. Apoptosis of oligodendrocyte precursors was determined by cleaved caspase-3 detection. Oligodendrocyte myelination was assessed by immunofluorescence, while microglia and astrocyte proinflammatory activation were assessed by both immunofluorescence and RT-qPCR. Methadone treatment increased neuronal apoptosis and reduced synaptic density. Furthermore, it led to increased oligodendrocyte apoptosis and a reduction in the myelinating capacity of these cells, and promoted the proinflammatory activation of microglia and astrocytes. We showed that methadone, the most widely used drug in opioid replacement therapy for pregnant women with opioid addiction, directly impairs brain cells in vitro, highlighting the need for developing alternative therapies to address opioid addiction in this population.

The axonal endoplasmic reticulum: One organelle-many functions in development, maintenance, and plasticity.

The endoplasmic reticulum (ER) is highly conserved in eukaryotes and neurons. Indeed, the localization of the organelle in axons has been known for nearly half a century. However, the relevance of the axonal ER is only beginning to emerge. In this review, we discuss the structure of the ER in axons, examining the role of ER-shaping proteins and highlighting reticulons. We analyze the multiple functions of the ER and their potential contribution to axonal physiology. First, we examine the emerging roles of the axonal ER in lipid synthesis, protein translation, processing, quality control, and secretory trafficking of transmembrane proteins. We also review the impact of the ER on calcium dynamics, focusing on intracellular mechanisms and functions. We describe the interactions between the ER and endosomes, mitochondria, and synaptic vesicles. Finally, we analyze available proteomic data of axonal preparations to reveal the dynamic functionality of the ER in axons during development. We suggest that the dynamic proteome and a validated axonal interactome, together with state-of-the-art methodologies, may provide interesting research avenues in axon physiology that may extend to pathology and regeneration. © 2017 Wiley Periodicals, Inc. Develop Neurobiol 78: 181-208, 2018.

Hipertensión en relación con estado nutricional, actividad física y etnicidad en niños chilenos entre 6 y 13 años de edad.

 Introducción: obesidad e inactividad física son importantes factores de riesgo para el desarrollo de hipertensión en adultos. No obstante, hay poca evidencia sobre el efecto de estos factores de riesgo en el desarrollo de hipertensión en población infantil. Objetivo: investigar la asociación del estado nutricional, niveles de actividad física y etnicidad con niveles de hipertensión en escolares entre 6 y 13 años de edad. Métodos: un total de 418 escolares de ascendencia étnica europea (n = 311) y mapuche (n = 107) fueron participantes de este estudio transversal. Se midió el peso, talla, índice de masa corporal (IMC) y presión arterial, utilizando protocolos estandarizados. Resultados: no se encontraron diferencias significativas en IMC, estado nutricional y presión arterial entre niños con ascendencia europea y mapuche. No obstante, la prevalencia de prehipertensión (21,3% vs. 11,1%) e hipertensión (28,9% vs. 18,6%) fue significativamente mayor en escolares mapuches en comparación con europeos, respectivamente. Escolares con ascendencia mapuche tienen un mayor riesgo de desarrollar prehipertensión o hipertensión que escolares con ascendencia europea (OR: 1,92 [1,19 a 3,06], p < 0,01). La prevalencia de hipertensión aumenta significativamente en ambos grupos étnicos con el incremento de IMC y bajos niveles de actividad física. Conclusiones: el riesgo de desarrollar prehipertensión o hipertensión es mayor en población infantil ascendiente de mapuches que ascendiente de europeos y este riesgo se ve acentuado con el incremento de obesidad y bajos niveles de actividad física.

Hipertensión en relación con estado nutricional, actividad física y etnicidad en niños chilenos entre 6 y 13 años de edad / Hypertension in relation to nutritional status, physical activity and ethnicity in Chilean children aged 6 to 13

Introducción: obesidad e inactividad física son importantes factores de riesgo para el desarrollo de hipertensión en adultos. No obstante, hay poca evidencia sobre el efecto de estos factores de riesgo en el desarrollo de hipertensión en población infantil. Objetivo: investigar la asociación del estado nutricional, niveles de actividad física y etnicidad con niveles de hipertensión en escolares entre 6 y 13 años de edad. Métodos: un total de 418 escolares de ascendencia étnica europea (n = 311) y mapuche (n = 107) fueron participantes de este estudio transversal. Se midió el peso, talla, índice de masa corporal (IMC) y presión arterial, utilizando protocolos estandarizados. Resultados: no se encontraron diferencias significativas en IMC, estado nutricional y presión arterial entre niños con ascendencia europea y mapuche. No obstante, la prevalencia de prehipertensión (21,3% vs. 11,1%) e hipertensión (28,9% vs. 18,6%) fue significativamente mayor en escolares mapuches en comparación con europeos, respectivamente. Escolares con ascendencia mapuche tienen un mayor riesgo de desarrollar prehipertensión o hipertensión que escolares con ascendencia europea (OR: 1,92 [1,19 a 3,06], p < 0,01). La prevalencia de hipertensión aumenta significativamente en ambos grupos étnicos con el incremento de IMC y bajos niveles de actividad física. Conclusiones: el riesgo de desarrollar prehipertensión o hipertensión es mayor en población infantil ascendiente de mapuches que ascendiente de europeos y este riesgo se ve acentuado con el incremento de obesidad y bajos niveles de actividad física (AU)

Introduction: Obesity and physical inactivity are two main risk factors for developing hypertension (HTA) in adulthood. However, there is a lack of evidence regarding the effect of this risk factor on HTA in childhood. Objective: The aim of this study was to investigate the association between nutritional status, physical activity levels and ethnicity with hypertension in scholar aged between 6 and 13 years. Methods: 418 school children from European (n = 311) and Mapuches (n = 107) background were invited to take part in the study. Height, weight, body mass index (BMI) and blood pressure were measured with standardised protocols. Results: No significant differences were found for IMC, nutritional status, blood pressure between European and Mapuches children. However, the prevalence of pre-HTA (21.3% vs. 11.1%) and HTA (28.9% vs. 18.6%) were significantly higher in Mapuches than European. Similarly, Mapuches are more likely to develop pre-HTA and HTA than Europeans children [OR: 1.92 (1.19 a 3.06) p < 0.010]. The prevalence of HTA was associated with BMI and physical activity, but not differences were found between ethnic groups. Conclusions: The risk of developing pre-HTA or HTA is higher in Mapuches than European children, and this risk increase with higher levels of BMI and lower levels of physical activity (AU)