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障碍 (FASD) 每年影响美国多达 5% 的新生儿，并导致终身身体和行为障碍。神经病学阶段的乙醇暴露（人类怀孕第 4 周，小鼠妊娠第 8-10 天），与面部和大脑的扩大有关，特别是腹侧中线结构（例如隔膜、垂体、心室）和生命后期的神经功能变化。类似于在朱伯特氏病等纤毛病遗传性疾病中观察到的中枢神经系统和颅面异常纤毛病是初级纤毛缺陷的结果，初级纤毛是对感觉细胞至关重要的不动性细胞器。发育过程中的音刺猬 (Shh) 通路转导和细胞增殖。一些纤毛病会导致 Shh 过度激活，从而导致先前研究中观察到的中枢神经系统异常。表明纤毛相关运动蛋白的突变会导致纤毛病表型并与乙醇导致更宽的大脑（未发表的数据）在这个提案中，我们使用了一个特征良好的小鼠模型。 FASD 的目的是检验神经溶解过程中乙醇暴露会诱发“短暂”纤毛病的假设胚胎，导致纤毛病和 FASD 的共同表型目标 1 初级纤毛数量。目标 2 研究了神经溶解阶段乙醇暴露后的神经管形态。通过检查纤毛相关基因表达变化来了解这些纤毛缺陷的机制最后，目标 3。通过分析检查神经溶解阶段的乙醇暴露是否会改变纤毛的稳定性和功能微管蛋白翻译后修饰和 Shh 通路信号传导。暴露的胚胎改变了关键纤毛发生基因的表达，并且增加了纤毛数量。更多纤毛的存在会导致 Shh 信号传导上调和细胞增殖异常，重要的是，这些实验将提供支持证据。神经管中初级纤毛数量和功能的改变作为乙醇的新途径暴露会导致 FASD 症状。