Primary cilia number and function in the neural tube in a mouse model of FASD

FASD 小鼠模型神经管初级纤毛的数量和功能

基本信息

批准号：
9469089
负责人：
Karen Elizabeth Boschen
金额：
$ 5.67万
依托单位：
UNIV OF NORTH CAROLINA CHAPEL HILL
依托单位国家：
美国
项目类别：
财政年份：
2018
资助国家：
美国
起止时间：
2018-03-15 至 2020-03-14
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9469089
关键词：
Abnormal Cell Acetylation Affect Behavioral Biological Assay Birth Brain Cell Proliferation Child Cilia Cleaved cell Cognitive Confocal Microscopy Corpus Callosum Craniofacial Abnormalities Data Defect Development Digit structure Dysmorphology Embryo Ethanol Etiology Event Exhibits Eye Face Failure Fetal Alcohol Exposure Fetal Alcohol Spectrum Disorder Fetal Alcohol Syndrome Fetus First Pregnancy Trimester Functional disorder Gene Expression Gene Targeting Genes Genetic Genetic Transcription Goals Hereditary Disease Holoprosencephaly Human Image Joubert syndrome Lead Length Life Link Lip structure Live Birth Mediating Microtubules Modification Molecular Morphology Motor Mus Mutant Strains Mice Mutation Neural tube Orbital separation excessive Organelles Palate Pathogenesis Pathway interactions Phenotype Pituitary Gland Population Post-Translational Protein Processing Pregnancy Process Prosencephalon Proteins Quantitative Reverse Transcriptase PCR RNA Reporting SHH gene Sensory Signal Pathway Signal Transduction Sonic Hedgehog Pathway Structure Symptoms Testing Thinness Time Tissue Expansion Tissues Translations Tubulin Up-Regulation Western Blotting Work alcohol exposure base behavioral impairment brain abnormalities ciliopathy cilium biogenesis craniofacial experimental study gastrulation hedgehog signal transduction knock-down mouse model novel prenatal reconstruction smoothened signaling pathway therapeutic target transcriptome sequencing

项目摘要

Abstract Fetal Alcohol Spectrum Disorders (FASD) affects up to 5% of births in the US each year and results in life-long physical and behavioral impairments. Neurulation-stage ethanol exposure (~ 4th week of pregnancy in humans, gestational days 8-10 in mice), is associated with a widening of the face and brain, particularly the ventral midline structures (e.g. septum, pituitary, ventricles), and neurofunctional changes later in life. This expansion is similar to the CNS and craniofacial abnormalities observed in ciliopathic genetic disorders such as Joubert’s syndrome. Ciliopathies are a consequence of defects in primary cilia, immotile sensory organelles critical for sonic hedgehog (Shh) pathway transduction and cell proliferation during development. Cilia dysfunction in some ciliopathies results in an overactivation of Shh, leading to the observed CNS anomalies. Previous work has suggested that mutations in cilia-associated motor proteins lead to ciliopathic phenotypes and interact with ethanol to cause wider brains (unpublished data). In this proposal, we use a well-characterized mouse model of FASD to test the hypothesis that ethanol exposure during neurulation induces a “transient” ciliopathy in the embryo, leading to the shared phenotype between ciliopathies and FASD. Aim 1 analyzes primary cilia number and morphology in the neural tube following neurulation-stage ethanol exposure. Aim 2 investigates the mechanisms of these ciliary defects by examining cilia-related gene expression changes. Finally, Aim 3 examines whether neurulation-stage ethanol exposure alters cilia stability and function through analysis of tubulin post-translational modifications and Shh pathway signaling. Preliminary data suggest that ethanol- exposed embryos have altered expression of key ciliogenesis genes and an increased number of cilia in the neural tube. Presence of more cilia would lead to upregulated Shh signaling and abnormal cell proliferation, causing the observed CNS abnormalities. Importantly, these experiments will provide evidence supporting alterations to primary cilia number and function in the neural tube as a novel pathway through which ethanol exposure causes symptoms of FASD.

抽象的胎儿酒精谱系（FASD）每年最多影响美国5％的出生，并导致终身身体和行为障碍。神经阶段乙醇暴露（人类怀孕的第四周，小鼠的妊娠天8-10），与面部和大脑的扩大有关中线结构（例如，隔膜，倍感，心室）和神经功能的变化以后生活。这个扩展类似于CNS和颅面异常，在纤毛性遗传疾病（例如Joubert的）中观察到综合征。纤毛病是原发性纤毛缺陷的结果发育过程中的Sonic刺猬（SHH）途径翻译和细胞增殖。纤毛功能障碍一些纤毛病导致SHH过度活化，导致观察到的中枢神经系统异常。以前的工作已经表明，与纤毛相关的运动蛋白中的突变导致纤毛性表型并与乙醇引起更广泛的大脑（未发表的数据）。在此提案中，我们使用特征良好的鼠标模型 FASD检验以下假设：神经化过程中乙醇暴露会诱导“短暂”纤毛病。胚胎，导致纤毛病和FASD之间的共同表型。 AIM 1分析初级纤毛数神经元乙醇暴露后神经元管中的形态。 AIM 2调查这些纤毛缺陷的机制通过检查与纤毛相关的基因表达变化。最后，目标3 检查神经化阶段乙醇暴露是否通过分析来改变纤毛的稳定性和功能微管蛋白翻译后修饰和SHH途径信号传导。初步数据表明乙醇 - 裸露的胚胎已经改变了关键纤毛生成基因的表达，并且纤毛数量增加神经管。存在更多的纤毛会导致更新的SHH信号传导和异常细胞增殖，导致观察到的中枢神经系统异常。重要的是，这些实验将提供支持的证据在神经管中的原发性纤毛数和功能的改变，作为一种新的途径，乙醇通过暴露会导致FASD症状。