Astrocyte-neuron interactions and sulfatases in Fetal Alcohol Spectrum Disorders

胎儿酒精谱系障碍中的星形胶质细胞-神经元相互作用和硫酸酯酶

• 批准号: 9297181
• 负责人: Marina Guizzetti
• 金额: $ 28.74万
• 依托单位: OREGON HEALTH & SCIENCE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-09-05 至 2020-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9297181
• 关键词: Affect; Antioxidants; Architecture; Arylsulfatase B; Astrocytes; Axon; Behavior; Behavioral; Biological; Brain; CSPG3 gene; Chondroitin Sulfate A; Chondroitin Sulfate C; Chondroitin Sulfate Proteoglycan; Clinical Research; Coculture Techniques; Cognitive; Communication; Core Protein; Cues; Dendrites; Development; Disaccharides; Enzyme Activation; Enzymes; Ethanol; Experimental Models; Family; Fetal Alcohol Spectrum Disorder; GAG Gene; Galactose; Generations; Hippocampus (Brain); Hypoxia; In Vitro; Injection of therapeutic agent; Investigation; Learning; Length; Link; Measures; Memory; Memory impairment; Messenger RNA; National Institute on Alcohol Abuse and Alcoholism; Neonatal; Neonatal Alcohol Exposure; Neurites; Neuronal Plasticity; Neurons; Oxidative Stress; Performance; Prevention; Property; Proteoglycan; Rattus; Reactive Oxygen Species; Recombinants; Reporting; Research; Role; Side; Signal Transduction; Small Interfering RNA; Sulfatases; Supplementation; Testing; Unspecified or Sulfate Ion Sulfates; Vertebral column; Work; alcohol abuse therapy; alcohol effect; alcohol exposure; brain abnormalities; brevican; chondroitin sulfate glycosaminoglycan; density; effective therapy; hippocampal pyramidal neuron; in vivo; innovation; insight; liquid chromatography mass spectrometry; molecular targeted therapies; morris water maze; neuron development; novel; novel therapeutic intervention; permissiveness; postnatal; preclinical study; prevent; public health relevance; versican

 DESCRIPTION (provided by applicant): Fetal Alcohol Spectrum Disorders (FASD) are characterized by structural brain abnormalities and compromised cognitive and behavioral functions. Neuronal connectivity and plasticity are affected by developmental alcohol exposure. The exposure of astrocytes to ethanol inhibits neuritogenesis in hippocampal neurons co-cultured with ethanol-treated astrocytes. However, a lack of understanding of the mechanisms by which ethanol affects neuronal structural plasticity in the developing brain persists. Filling tis gap will open the path to the identification of highly effective treatments to prevent or ameliorat the developmental effects of ethanol. Lecticans are inhibitory proteoglycans that, in the brain, prevent the extension of axons and dendrites. Lecticans consist of a core-protein moiety covalently bound to linear chains of disaccharides, chondroitin sulfates glycosaminoglycans (CS-GAGs), which are comprised mostly of chondroitin-4 sulfate (C4S) and chondroitin-6 sulfate (C6S); the inhibitory properties of lecticans depend on their core-protein and their sGAG side-chains. Two sulfatase enzymes, arylsulfatase B (ARSB) and galactose-6-sulfatase (GALNS) remove sulfate groups from C4S and C6S respectively and are required for the degradation of CS-GAGs. We find that ethanol inhibits the activity of ARSB and GALNS and that ethanol and ARSB silencing increase the expression of the lectican neurocan and C4S-GAG content in astrocytes in vitro. ARSB activity is inhibited, neurocan and sGAG levels are upregulated, and pyramidal neuron dendritic arborization is reduced also in the hippocampus of neonatal rats after in vivo ethanol exposure. Reduced ARSB activity and dendritic arborization persist up to post-natal day 36 and is correlated with reduced performance in spatial learning and memory tasks. In this proposal we hypothesize that the inhibition of ARSB and GALNS activity by ethanol in the developing hippocampus induces an unscheduled increase in lectican (neurocan, brevican, and versican) core-protein and CS-GAG levels leading to decreased in neuronal structural plasticity; these effects are long-lasting and causally linked to reduced performance in learning and memory tasks and are rescued by recombinant (r)ARSB. We plan to test our hypothesis by pursuing the following three specific aims: 1) To investigate the mechanisms of ethanol-induced sulfatase inhibition and the consequences on lectican core-proteins and sGAG levels and on CS disaccharide levels and composition in astrocytes. 2) To investigate the role of: sulfatases, SUMF1, lecticans, and oxidative stress on ethanol-treated astrocyte inhibition of neurite outgrowth. 3) To investigate the effects of in vivo neonatal ethanol exposure and of rARSB injections on sulfatase activity, lectican expression, sGAG levels and CS disaccharide levels, and dendritic arborization in the hippocampus and on spatial learning and memory. The proposed work is innovative and significant as it explores a novel mechanism underlying ethanol-induced inhibition of neuronal plasticity and will yield strategies aimed at sulfatase enzyme activation for the treatment of alcohol teratogenesis therefore advancing FASD research.

 描述（由适用提供）：胎儿酒精谱系障碍（FASD）的特征是结构性脑部异常和认知和行为功能受损。神经元连通性和可塑性受到发育性酒精暴露的影响。星形胶质细胞暴露于乙醇中，抑制与乙醇处理的星形胶质细胞共培养的海马神经元中的神经发生。但是，缺乏对乙醇影响发育中大脑中神经元结构可塑性的机制的理解。填充间隙将为鉴定高效处理以预防或改善乙醇的发育效果开辟道路。固定剂是抑制性蛋白聚糖，在大脑中可以防止轴突和树突的延伸。拉克斯人由核蛋白部分组成，共价结合到二糖，软骨素硫酸盐糖胺聚糖（CS-GAGS）的线性链中，这些链蛋白硫酸盐糖糖糖糖糖糖糖糖糖糖糖糖糖糖糖蛋白（CS-GAGS）主要由软骨素-4硫酸盐（C4S）和软骨素-6硫酸盐（C6S）组成；拉克剂的抑制特性取决于其核蛋白及其SGAG侧链。两种硫酸酶B（ARSB）和半乳糖-6-硫酸酶（GALNS）分别从C4和C6S中去除硫酸盐基团，这是CS-GAG降解所必需的。我们发现乙醇抑制了ARSB和GALN的活性，并且在体外星形胶质细胞中，乙醇和ARSB沉默会增加拉克斯神经胶质和C4S-GAG含量的表达。 ARSB活性受到抑制，神经can和SGAG水平被上调，在体内乙醇暴露后，在新生大鼠的海马中也降低了锥体神经元树突状树皮化。降低的ARSB活性和树突状树博化持续到产后第36天，与空间学习和记忆任务的性能降低相关。在这项提案中，我们假设乙醇在发育中的海马中抑制ARSB和GALNS活性会影响拉克（Neurocan，Brevican和Brevican和Versican）核心蛋白和CS-GAG的固定剂降低，导致神经元结构可塑性下降，导致神经元核蛋白和CS-GAG水平降低；这些效果是长期的，并且与降低的性能有关 学习和记忆任务，并由重组（R）ARSB做出响应。我们计划通过追求以下三个特定目的来检验假设：1）研究乙醇诱导的硫酸糖抑制的机制，以及对胶状核蛋白和SGAG水平以及CS脱糖水平以及星形胶质细胞中CS脱糖水平以及组成的后果。 2) To investigate the role of: sulfatases, SUMF1, lecticans, and oxidative stress on ethanol-treated astrocyte inhibition 3) To investigate the effects of in vivo neonatal ethanol exposure and of rARSB injections on sulfatase activity, lectican expression, sGAG levels and CS disaccharide levels, and dendritic arborization in the hippocampus and on spatial learning and 记忆。拟议的工作具有创新性和意义，因为它探索了乙醇诱导的神经元可塑性抑制的一种新型机制，并将产生针对硫酸酶酶激活的策略，以治疗酒精致病发生的治疗。