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: 8887358
• 负责人: Kassandra Marie Ori-McKenney
• 金额: $ 9.14万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-07-10 至 2016-01-03
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8887358
• 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; 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(申请人提供)：树突场的图案和形态决定了神经元能够接受的输入的数量和类型。因此，树突缺陷是唐氏综合症(DS)等遗传性疾病中表现出的智能障碍的最强解剖相关性。在最近对果蝇树突状分支神经元树突异常的筛查中，我发现迷你脑(MNB)激酶是树枝形成过程中细胞骨架的重要调节因子。MNB是DS的关键基因，与神经元发育、脑功能和DS病理密切相关。然而，目前尚不清楚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。这项建议的目的是回答这些问题，并确定MNB激酶在树突形成过程中的机制作用，以了解这一途径如何促进神经元的发育和功能。在哥伦比亚大学理查德·瓦莱博士的研究生培训期间，我将体外生化和生物物理分析与体内神经元技术相结合，以破译分子马达和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。在加州大学旧金山分校Jan博士的博士后生涯中，我在果蝇遗传学、分子生物学和完整幼虫的活体成像方面积累了其他技能，以研究在神经元中创建特定树突树枝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，我将与每个领域的专家合作，获得行为分析、质谱学和活体超分辨率显微镜方面的额外培训。这项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)利用 化学遗传学方法，2)利用STORM分析MNB及其底物在控制和类似DS条件下树突发育和维持过程中的功能，以及3)描述果蝇学习和记忆中MNB途径的特征。随着在K99阶段获得的新培训和信息，我将扩大我在R00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磷酸化靶标。此外，我计划研究TH小脑磷酸化靶标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。总之，这一建议的实施将有助于阐明神经元功能和功能障碍过程中树突形成的途径，并有助于我们理解MNB在DS进展中的机制。