Engrailed genes and cerebellum morphology, spatial gene expression and circuitry

纠缠基因和小脑形态、空间基因表达和电路

• 批准号: 8777059
• 负责人: ALEXANDRA L. JOYNER
• 金额: $ 63.94万
• 依托单位: SLOAN-KETTERING INST CAN RESEARCH
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-04-01 至 2018-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8777059
• 关键词: Address; Adhesions; Affect; Affective; Afferent Neurons; Afferent Pathways; Architecture; Autistic Disorder; Back; Behavior; Behavioral; Binding; Biochemical; Brain; CCL4 gene; Cell Count; Cell Cycle Regulation; Cell Lineage; Cell Proliferation; Cell Survival; Cell division; Cell physiology; Cells; Cerebellar Nuclei; Cerebellum; ChIP-seq; Complex; Cytoplasmic Granules; Defect; Development; Developmental Gene; Disease; Embryo; Emotional; Engineering; Ensure; Funding; Gene Expression; Gene Expression Profile; Gene Targeting; Genes; Genetic; Genetic Processes; Genetic Techniques; Growth; Health; Homeobox; Human; Image; In Vitro; Knockout Mice; Language; Life; Lip structure; Methods; Microarray Analysis; Molecular; Morphogenesis; Morphology; Motor; Mouse Strains; Mus; Mutant Strains Mice; Mutate; Neurons; Output; Pathway Analysis; Pathway interactions; Patients; Pattern; Performance; Pharmaceutical Preparations; Phenotype; Play; Positioning Attribute; Predisposition; Process; Production; Proteins; Purkinje Cells; Rest; Role; Slice; Social Behavior; Social Functioning; Sonic Hedgehog Pathway; Techniques; Testing; Time; autism spectrum disorder; base; behavioral study; cell behavior; cell type; cellular imaging; cognitive function; comparative; differential expression; disease diagnosis; feeding; in vivo; insight; migration; motor control; mutant; nerve stem cell; novel; public health relevance; research study; smoothened signaling pathway; social; social cognition; transcription factor

DESCRIPTION (provided by applicant): The cerebellum (Cb) is implicated in contributing to cognitive and social functions, in addition to having a critical role in skilled motor performance. Accordingly, the Cb is associated with many debilitating developmental diseases including autism. One gene that regulates development of the Cb and has been implicated in autism is engrailed 2 (EN2), based on human studies and the finding that En2 null mice not only have deficits in motor control, but also in social behaviors and cognition. Before we can begin to understand higher order functions of the Cb, we must gain more insight into the basic cellular and genetic processes that regulate Cb development. Our approach is to use the two EN homeobox transcription factors as molecular entry points to study Cb development, as we discovered that En1/2 conditional mutants have defects in Cb morphology, molecular patterning and afferent circuitry. We will now direct our studies towards distinguishing the cellular processes regulated by En1/2 and identifying EN2 target genes critical for these processes that could be susceptibility loci for complex behavioral diseases. We will focus on the granule neurons (GNs) that comprise the main recipients of input to the Cb and the deep cerebellar nuclei (DCN) that generate the output. DCN neurons are consistently reduced in autistic patients, which could be a primary cause of some behaviors and also reflect defects elsewhere in the Cb circuit. We will apply a multi-facetted approach that combines novel genetic techniques in mice to study normal and mutant behaviors of GNs and DCN projection neurons, including a mosaic mutant analysis using our MASTR technique and a new method to precisely target over- expression of EN2 to GNs and the DCN to test sufficiency of EN2 to alter differentiation. We will then apply both mutant approaches to live imaging of GNs as a different approach to study Cb morphogenesis and cell proliferation/differentiation. We will also address the question of whether feed back loops ensure the correct proportion of cell types is produced by studying the interaction between En1/2 and the sonic hedgehog (SHH) pathway. Finally, in order to identify the first direct targets of EN2 in the brain, we are engineering new mouse strains expressing a tagged form of EN2. Aim 1. Study the cellular behaviors regulated by En1/2 in developing GNs and DCN projection neurons using conditional genetics and characterizing cellular behaviors in vivo, and in vitro with live imaging. Aim 2. Identify critical target genes o EN1/2 in GN precursors and DCN projection neurons using comparative microarray analysis and ChIP-seq.

描述(由申请人提供)：小脑(CB)除了在熟练的运动表现中起关键作用外，还与认知和社会功能有关。 因此，CB与包括自闭症在内的许多衰弱的发育疾病有关。调节CB发育的一个基因是Engraile2(EN2)，它与自闭症有关，这是基于人类研究和发现EN2基因缺失的小鼠不仅在运动控制方面存在缺陷，而且在社会行为和认知方面也存在缺陷。在我们开始了解CB的高阶功能之前，我们必须更深入地了解调控CB发育的基本细胞和遗传过程。我们的方法是使用两个EN同源框转录因子作为分子切入点来研究CB的发育，因为我们发现EN1/2条件突变在CB的形态、分子构型和传入回路方面存在缺陷。我们现在将引导我们的研究，以区分EN1/2调控的细胞过程，并识别对这些过程至关重要的EN2靶基因，这些基因可能是复杂行为疾病的易感基因。我们将专注于颗粒神经元(GN)，它包括CB输入的主要接受者和产生输出的小脑深核(DCN)。自闭症患者的DCN神经元持续减少，这可能是某些行为的主要原因，也反映了CB回路的其他方面的缺陷。我们将应用多方面的方法，结合新的遗传技术在小鼠身上研究GNS和DCN投射神经元的正常和突变行为，包括使用我们的MASTR技术进行镶嵌突变分析，以及一种新的方法来精确定位EN2在GNS和DCN的过度表达，以测试EN2是否足以改变分化。然后，我们将应用这两种突变方法对肾小球进行实时成像，作为研究CB形态发生和细胞增殖/分化的不同方法。我们还将通过研究EN1/2和Sonic Hedgehog(SHH)途径之间的相互作用来解决反馈环是否确保产生正确比例的细胞类型的问题。最后，为了确定EN2在大脑中的第一个直接靶点，我们正在设计表达一种标记形式的EN2的新的小鼠品系。目的1.用条件遗传学方法研究EN1/2对GNS和DCN投射神经元发育过程中细胞行为的调控，并通过活体和体外图像分析细胞行为特征。目的2.利用比较微阵列技术和芯片序列分析技术，鉴定GN前体和DCN投射神经元中的EN1/2关键靶基因。