Roles of microRNAs during vertebrate brain morphogenesis

microRNA 在脊椎动物脑形态发生中的作用

• 批准号: 8262329
• 负责人: Carter M. Takacs
• 金额: $ 5.3万
• 依托单位: YALE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-06-01 至 2013-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8262329
• 关键词: Anencephaly; Base of the Brain; Behavior; Biogenesis; Bioinformatics; Brain; Candidate Disease Gene; Cell Adhesion; Cell Lineage; Cell Polarity; Cell division; Cell physiology; Cell-Cell Adhesion; Cells; Cellular Morphology; Cerebral Ventricles; Cerebral cortex; Characteristics; Confocal Microscopy; Defect; Development; Dicer Enzyme; Disease; Economic Inflation; Embryo; Ensure; Epilepsy; Epithelium; Excision; Failure; Functional RNA; Gene Expression; Gene Targeting; Genes; Genetic; Genetic Screening; Genetic Translation; High Prevalence; Human; Image; Individual; Knowledge; Label; Laboratories; Life; Medial; Mental Health; Messenger RNA; MicroRNAs; Microarray Analysis; Molecular; Monitor; Morphogenesis; Morphology; Movement; Mus; Nature; Nervous System Physiology; Neural Tube Defects; Neural tube; Neuroectodermal Cell; Neurons; Normal tissue morphology; Organogenesis; Pathway interactions; Pattern; Phenotype; Physiological; Process; Production; Property; Regulation; Regulator Genes; Role; Seeds; Series; Shapes; Spinal; Spinal Cord; Spinal Dysraphism; Techniques; Time; Tissues; Translations; Zebrafish; base; brain morphology; cell fate specification; cell motility; cell type; developmental neurobiology; in vivo; insight; mRNA Stability; mRNA Transcript Degradation; malformation; mutant; neural plate; novel; public health relevance; relating to nervous system; research study; ventricular system

DESCRIPTION (provided by applicant): A fundamental question in developmental neurobiology is how the vertebrate brain acquires its characteristic morphology to facilitate neuronal connectivity and organization. MicroRNAs (miRNAs) have recently emerged as novel regulators of gene expression with potential widespread influence on development and disease. To investigate the roles of miRNAs, the Giraldez laboratory has generated mutant zebrafish embryos (MZdicer) that are devoid of miRNA function. MZdicer embryos display normal patterning but fail to undergo proper neural tube morphogenesis and brain ventricle formation. Reintroduction of an early expressed miRNA (processed miR- 430) rescues these defects, indicating that miR-430 regulation is critical for proper morphogenesis. This proposal aims to determine how miR-430 governs gene expression to shape cell movement and organization during brain morphogenesis. Experiments outlined in Aim #1 will provide a detailed characterization of the MZdicer neural phenotype by combining time-lapse confocal microscopy with single-cell labeling and genetic mosaic analyses. These experiments will elucidate the onset and nature of defects in cell movement and organization that underlie the MZdicer neural phenotype. Experiments outlined in Aim #2 will identify the miR-430 target genes whose regulation is critical for proper brain morphogenesis. Time-course microarray and bioinformatic analyses will isolate mRNAs upregulated in the absence of miRNA function. Further, the in vivo functional relevance of individual miR-430-target mRNA interactions will be assessed through a combination of target misexpression and protection techniques. Taken together, these experiments will provide mechansitic insights into how miRNA activity instructs cellular behaviors during vertebrate brain morphogenesis. PUBLIC HEALTH RELEVANCE: Genetic malformations of the cerebral cortex cause severe mental health impediments, including retardation and epilepsy. In addition, failure of brain closure (anencephaly) and spinal closure (spina bifida) occur with a high prevalence in humans with devastating effects to nervous system function. This study has the potential to increase our understanding of the genetic basis of brain and spinal cord malformations by elucidating the regulatory processes that instruct neural tube and brain morphology during vertebrate development.

描述（由申请人提供）：发育神经生物学中的一个基本问题是脊椎动物大脑如何获得其特征形态以促进神经元的连通性和组织。 MicroRNA（miRNA）最近成为基因表达的新调节剂，对发育和疾病的潜在影响。为了研究miRNA的作用，吉拉尔德斯实验室产生了不含miRNA功能的突变斑马鱼胚胎（MZDICER）。 MZDICER胚胎表现出正常的模式，但无法经历适当的神经管形态发生和脑室心室形成。重新引入早期表达的miRNA（加工miR-430）挽救了这些缺陷，表明miR-430调控对于正确的形态发生至关重要。该建议旨在确定miR-430如何控制基因表达以塑造脑形态发生过程中细胞运动和组织。 AIM＃1中概述的实验将通过将延时共聚焦显微镜与单细胞标记和遗传镶嵌分析相结合，从而提供MZDICER神经表型的详细表征。这些实验将阐明MZDICER神经表型的细胞运动和组织中缺陷的开始和性质。 AIM＃2中概述的实验将识别MiR-430靶基因，其调节对于适当的脑形态发生至关重要。时间课微阵列和生物信息学分析将在没有miRNA功能的情况下隔离mRNA。此外，将通过目标misexpression和保护技术的组合来评估单个miR-430目标mRNA相互作用的体内功能相关性。综上所述，这些实验将提供有关miRNA活性在脊椎动物脑形态发生过程中如何指导细胞行为的机制见解。 公共卫生相关性：大脑皮层的遗传畸形会导致严重的心理健康障碍，包括迟缓和癫痫。另外，脑闭合（肛门颅）和脊柱闭合（脊柱裂）的失败发生，人类的患病率很高，对神经系统功能具有毁灭性影响。这项研究有可能通过阐明指导脊椎动物发育过程中神经管和脑形态的调节过程来增强我们对大脑和脊髓畸形的遗传基础的理解。