Role of the Down Syndrome gene, minibrain, in neuronal development and function

唐氏综合症基因、小脑在神经元发育和功能中的作用

• 批准号: 9207204
• 负责人: Kassandra Marie Ori-McKenney
• 金额: $ 24.9万
• 依托单位: UNIVERSITY OF CALIFORNIA AT DAVIS
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-01-04 至 2019-01-03
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9207204
• 关键词: Actins; Affect; Behavioral; Behavioral Assay; Biochemical; Biochemistry; Biological Assay; Brain; California; Chromosomes, Human, Pair 21; Collaborations; Conflict (Psychology); Cytoskeletal Modeling; Cytoskeleton; Data; Defect; Dendrites; Development; Disease; Down Syndrome; Drosophila genus; Dynein ATPase; Enhancers; Fragile X Syndrome; Functional disorder; Gene Dosage; Genes; Genetic; Goals; Health; Hereditary Disease; Homologous Gene; Human; Image; In Vitro; Intellectual functioning disability; Laboratory Research; Larva; Lead; Learning; Learning Disabilities; Life; Maintenance; Mass Spectrum Analysis; Memory; Mentors; Methods; Microscopy; Microtubules; Molecular; Molecular Biology; Molecular Motors; Morphology; Mus; Mutation; Nerve Degeneration; Neurologic; Neurons; Optics; Pathology; Pathway interactions; Pattern; Phase; Phenotype; Phosphorylation; Phosphorylation Site; Phosphotransferases; Physiological; Recombinants; Research; Research Personnel; Resolution; Role; San Francisco; Staging; Syndrome; System; Techniques; Testing; Training; Universities; career; chemical genetics; classical conditioning; genetic approach; in vivo; insight; knock-down; long term memory; loss of function; neurodevelopment; neuron development; novel; overexpression; reconstruction; research study; skills

DESCRIPTION (provided by applicant): The patterning and morphology of a dendritic field determines the number and type of inputs that a neuron is able to receive. Consequently, dendrite defects are the strongest anatomical correlates of the intellectual disabilities manifeste in genetic disorders such as Down Syndrome (DS). In a recent screen for dendrite abnormalities in Drosophila dendritic arborization neurons, I identified minibrain (mnb) kinase as an important regulator of the cytoskeleton during arborization. Mnb is a DS critical gene that has been implicated in neuronal development, brain function and DS pathology. However, it is currently unclear what the physiological substrates of mnb are in neurons, what the molecular mechanism of mnb is during dendrite development and maintenance, and how the imbalance in mnb gene dosage might cause the neuronal and behavioral alterations associated with DS. The goal of this proposal is to answer these questions and define the mechanistic role of mnb kinase during dendrite formation, in order to understand how this pathway contributes to neuronal development and function. During my graduate training with Dr. Richard Vallee at Columbia University, I combined in vitro biochemical and biophysical assays with in vivo neuronal techniques in order to decipher how mutations in the molecular motor, cytoplasmic dynein, cause neurodegenerative and neurodevelopmental diseases in mice and humans. During my postdoctoral career in Dr. Yuh-Nung Jan's lab at the University of California, San Francisco, I have developed additional skills in Drosophila genetics, molecular biology, and live imaging in intact larvae to study the cytoskeletal organization required to create specific dendritic arborization patterns in neurons. For the experiments proposed in this application, I will continue to employ these skills, and I will acquire additional training in behavioral analysis, mass spectrometry, and in vivo super resolution microscopy in collaboration with experts from each field. This training will allow me to achieve the following aims and launch an independent research laboratory within two years: 1) Identification of direct novel targets of mnb kinase using a chemical genetic approach, 2) Functional analysis of mnb and its substrates during dendrite development and maintenance under control and DS-like conditions using STORM, and 3) Characterization of the mnb pathway in Drosophila learning and memory. With the new training and information gained in the K99 phase, I will extend the scope of my research in the R00 phase. I plan to analyze the cytoskeletal interactions and the mnb phosphorylation targets during different stages of dendrite development. In addition, I plan to investigate the roles of th minibrain phosphorylation targets in Drosophila learning and memory. Finally, I will perform an enhancer/suppression screen to identify the mechanisms regulating mnb expression and activity during neuronal development. Overall, the implementation of this proposal will elucidate the pathways involved in dendrite formation during neuronal function and dysfunction and will contribute to our understanding of the mechanism of mnb in the progression of DS.

描述（由申请人提供）：树突场的模式和形态决定了神经元能够接收的输入的数量和类型。因此，树突缺陷是遗传疾病如唐氏综合征（DS）中表现出的智力残疾的最强解剖学相关性。在最近的屏幕上的树突异常果蝇树突树枝状神经元，我确定minibrain（mnb）激酶作为一个重要的调节细胞骨架在树枝状。Mnb是DS的关键基因，其与神经元发育、脑功能和DS病理学有关。然而，目前还不清楚mnb在神经元中的生理底物是什么，mnb在树突发育和维持过程中的分子机制是什么，以及mnb基因剂量的不平衡如何导致与DS相关的神经元和行为改变。本研究的目的是回答这些问题，并明确mnb激酶在树突形成过程中的作用机制，以了解该通路如何促进神经元的发育和功能。在我与哥伦比亚大学的Richard Vallee博士一起进行研究生培训期间，我将体外生物化学和生物物理测定与体内神经元技术相结合，以破译分子马达细胞质动力蛋白的突变如何导致小鼠和人类的神经退行性和神经发育疾病。在我的博士后生涯中Yuh-Nung Jan博士在加州大学弗朗西斯科分校的实验室，我开发了果蝇遗传学，分子生物学和完整幼虫的实时成像的额外技能，以研究在神经元中创建特定树突状分支模式所需的细胞骨架组织。对于本申请中提出的实验，我将继续 为了运用这些技能，我将与来自各个领域的专家合作，在行为分析、质谱分析和体内超分辨率显微镜方面获得额外的培训。这次培训将使我能够实现以下目标，并在两年内建立一个独立的研究实验室：1）使用 化学遗传学方法，2）使用STORM在控制和DS样条件下对mnb及其底物在树突发育和维持期间的功能分析，以及3）果蝇学习和记忆中mnb途径的表征。通过K99阶段获得的新培训和信息，我将扩大R 00阶段的研究范围。我计划分析树突发育不同阶段的细胞骨架相互作用和mnb磷酸化靶点。此外，我计划研究th minibrain磷酸化靶点在果蝇学习和记忆中的作用。最后，我将进行增强子/抑制筛选，以确定在神经元发育过程中调节mnb表达和活性的机制。总体而言，这一建议的实施将阐明树突形成过程中神经元功能和功能障碍的途径，并将有助于我们了解的机制，mnb在DS的进展。