Effect of Ethanol on Cell Proliferation

乙醇对细胞增殖的影响

• 批准号: 8462175
• 负责人: Frank A. Middleton
• 金额: $ 34.74万
• 依托单位: UPSTATE MEDICAL UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1992
• 资助国家: 美国
• 起止时间: 1992-08-01 至 2015-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8462175
• 关键词: Activity Cycles; Adult; Affect; Affinity; Alcohols; Attention deficit hyperactivity disorder; Autistic Disorder; Behavior; Brain; Cell Count; Cell Cycle; Cell Cycle Kinetics; Cell Proliferation; Cell Proliferation Regulation; Cell Transplants; Cells; Cerebral cortex; Characteristics; Complement; Corpus Callosum; Defect; Development; Dorsal; Environmental Impact; Environmental Risk Factor; Equilibrium; Ethanol; Ethanol toxicity; Etiology; Event; Exposure to; Fetal Alcohol Exposure; Fetal Alcohol Spectrum Disorder; Functional disorder; Gene Silencing; Genes; Genetic; Genomics; Growth; Harvest; Heterotopic Transplantation; Homeobox Genes; In Vitro; Indium; Kinetics; Ligands; Live Birth; Medial; Mental Retardation; Mental disorders; Modeling; Mus; Neuraxis; Neurons; Neuropeptides; Pattern; Phase; Phenotype; Population; Process; Production; Proliferating; Proteins; Psyche structure; Regulation; Shapes; Site; Slice; Staging; Stem cells; Structure; System; Telencephalon; Testing; Transforming Growth Factor beta; Transforming Growth Factors; Transplantation; United States; Variant; Ventricular; alcohol effect; brain size; defined contribution; excitatory neuron; human TGFB1 protein; in vivo; in vivo Model; inhibitory neuron; insight; migration; nerve stem cell; novel; prenatal exposure; progenitor; receptor; response; stem cell fate; subventricular zone

Understanding the effects of ethanol on neural stem cells is critical to unraveling the etiological knots of deficits associated with fetal alcohol spectrum disorder (FASD). FASD is a compelling problem because it is a major cause of developmental mental dysfunction affecting ~2% of all live births (indeed, it is the chief cause of mental retardation in the USA), and it provides insights into the etiology of other (often co-morbid) mental health disorders (e.g., attention deficit hyperactivity disorder and autism). The composition and size of the brain are established by the proliferation of neural stem cells and by the numbers and lineage of the derivatives. Improper numbers or balance of neuronal subpopulations can underlie mental dysfunction. Thus, we will test the hypothesis that ethanol affects the cycling behavior and fates of cells generated in the developing central nervous system. Cerebral cortex is composed of two types of neurons: excitatory projection neurons (PNs) and inhibitory local circuit neurons (LCNs). These derive prenatally from two distinct proliferative regions. PNs come from the dorsal telencephalon which comprises two zones: the ventricular zone (VZ) and its derivative, the subventricular zone (SZ). Most cortical LCNs are generated in the ventral telencephalon, in the medial ganglionic eminence (MGE). The fates of VZ/SZ and MGE cells are defined in a two-step process. (1) It is decided whether the cells remain in the cycling population and (2) the phenotype (e.g., the type of neuron) is defined. During the previous period of support, we showed that transforming growth factor (TGF) beta1 is a key regulator of neural stem cell proliferation. The present project will explore the effects of ethanol on dynamics of cell proliferation and on the definition of cell fate. Specific Aims 1 and 2 will use an in vivo model to determine the effects of ethanol on the cycling activity (cell cycle kinetics and exit) and on the expression/activation of TGFbeta receptors by neural stem cells in the cortical proliferative zones. During their development, neural stem cells express a homeobox gene product(s), Pax6 and/or Tbr2. These proteins define the transition from (Pax6+) stem cells in the VZ to an (Tbr2+) intermediate progenitor cell stage. This transition is critical for the development of superficial cortex (e.g., the origin of callosal projections) which is a target of prenatal exposure to ethanol. Specific Aim 3 will use two types of cultures (organotypic slices that retain in vivo-like organization and lines of neural stem cells harvested from the VZ/SZ or MGE) to explore the effects of ethanol on TGFbeta1-regulated cell proliferation and fate decisions. In addition, we will identify genes that are up- and down-regulated and silenced (methylated) by ethanol and/or TGFbeta1. In Specific Aim 4, neural stem cells will be transplanted (homotopically or heterotopically) to determine the effects of ethanol and/or TGFbeta1 on genetic and environmental contributions to determining cycling behavior and to defining cell fate. In concert, the novel Aims will use three complementary models to gain critical insight into mechanisms defining cell proliferation and fate and the effects of ethanol on these critical factors.

了解乙醇对神经干细胞的影响对于揭开缺陷的病因结是至关重要的 与胎儿酒精谱系障碍（FASD）相关。 FASD是一个引人入胜的问题，因为它是主要的 发育性心理功能障碍的原因影响了所有活产的约2％（实际上，这是精神的主要原因 在美国的迟钝），它提供了有关其他（通常合并）心理健康障碍的病因的见解 （例如，注意缺陷多动障碍和自闭症）。建立大脑的组成和大小 通过神经干细胞的增殖以及衍生物的数量和谱系。数字不正确或 神经元亚群的平衡可以是精神功能障碍的基础。因此，我们将检验乙醇的假设 影响发育中心神经系统中产生的细胞的循环行为和命运。 大脑皮层由两种类型的神经元组成：兴奋性投影神经元（PNS）和抑制性局部 电路神经元（LCN）。这些从两个不同的增殖区域中得出。 PNS来自背侧 端脑包括两个区域：心室区（VZ）及其衍生物，室内区。 （SZ）。大多数皮质LCN在腹侧脑脑，内侧神经节隆起（MGE）中产生。 VZ/SZ和MGE细胞的命运是在两步过程中定义的。 （1）确定细胞是否保留 在循环种群中，（2）定义了表型（例如，神经元的类型）。在上一个阶段 在支持中，我们表明转化生长因子（TGF）beta1是神经干细胞增殖的关键调节剂。 本项目将探讨乙醇对细胞增殖动态的影响以及对 细胞命运。具体目的1和2将使用体内模型来确定乙醇对循环活性的影响 （细胞周期动力学和出口）以及皮质中神经干细胞TGFBETA受体的表达/激活 增殖区。在发育过程中，神经干细胞表达同型基因基因产物，PAX6和/或 TBR2。这些蛋白质定义了从VZ中的（PAX6+）干细胞到（TBR2+）中间祖细胞的过渡 细胞阶段。这种过渡对于浅表皮质的发展至关重要（例如，呼叫的预测的起源） 这是产前暴露于乙醇的靶标。特定目标3将使用两种类型的培养物（器官型切片 保留从VZ/SZ或MGE收获的神经干细胞的体内式组织和线条，以探索 乙醇对TGFBETA1调节的细胞增殖和命运决策的影响。此外，我们将确定 由乙醇和/或TGFBETA1上调和下调和沉默（甲基化）。在特定目标4中，神经茎 细胞将被移植（同量或异位），以确定乙醇和/或TGFBETA1对 遗传和环境对确定循环行为和定义细胞命运的贡献。 在协同中，小说的目标将使用三种互补模型来获得对定义机制的关键见解 细胞增殖和命运以及乙醇对这些关键因素的影响。

项目成果

Alterations in serum microRNA in humans with alcohol use disorders impact cell proliferation and cell death pathways and predict structural and functional changes in brain.

• DOI: 10.1186/s12868-015-0195-x
• 发表时间: 2015-09-05
• 期刊: BMC neuroscience
• 影响因子: 2.4
• 作者: Ignacio C;Hicks SD;Burke P;Lewis L;Szombathyne-Meszaros Z;Middleton FA
• 通讯作者: Middleton FA