Effects of DEAF1 on Neuronal Activity and Target Gene Expression.

DEAF1 对神经元活动和靶基因表达的影响。

基本信息

批准号：
8871064
负责人：
Philip J. Jensik
金额：
$ 18.44万
依托单位：
SOUTHERN ILLINOIS UNIVERSITY CARBONDALE
依托单位国家：
美国
项目类别：
财政年份：
2015
资助国家：
美国
起止时间：
2015-02-01 至 2017-01-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8871064
关键词：
Adaptive Behaviors Affect Amino Acid Substitution Area Biological Assay Brain C57BL/6 Mouse DNA DNA Binding DNA Binding Domain Defect Dominant-Negative Mutation EMSA Eukaryotic Initiation Factor-4G Gene Expression Gene Mutation Gene Targeting Genes Genetic Predisposition to Disease Genetic Transcription Hippocampus (Brain)Human Immunofluorescence Immunologic In Vitro Individual Intellectual functioning disability Knock-out Learning Link Long-Term Depression Long-Term Potentiation Measurable Memory Missense Mutation Molecular Mus Mutate Mutation Neuraxis Neurons Outcome Pathogenesis Point Mutation Population Proteins RNA Reporting Research Role Sampling Serotonin Receptor 5-HT1A Slice Testing Variant anxiety-like behavior autism spectrum disorder base chromatin immunoprecipitation cognitive ability cognitive function conditioned fear differential expression exome sequencing in vivo insight mutant neuron development novel particle promoter protein function protein protein interaction public health relevance research study transcription factor transcriptome sequencing

项目摘要

DESCRIPTION (provided by applicant): Intellectual disability is characterized by below-average cognitive ability leading to significant deficits in functioning and adaptive behaviors. Th majority of the unexplained intellectual disability cases are caused by de novo gene mutations. Four individuals with intellectual disabilities were found to have heterozygous de novo point mutations within the DNA binding domain of the transcription factor Deformed Epidermal Autoregulatory Factor-1 (DEAF1), and the resultant amino acid substitutions were shown to negatively affect DEAF1 protein function. Four additional DEAF1 variants have recently been identified from a de-identified group of samples from individuals with intellectual disabilities an autism spectrum disorders. These mutations also occur within the DNA binding domain, but the effects of these mutations on DEAF1 function are currently uncharacterized. Since DEAF1 haploin sufficiency has not been reported to result in intellectual disabilities, this suggests tha the mutant DEAF1 proteins likely exert dominant negative influences on wildtype DEAF1 function. The underlying mechanisms by which mutant DEAF1 proteins contribute to the pathogenesis producing the intellectual disabilities remain unclear. The hypothesis for this proposal is that mutations in the DEAF1 gene that diminish DEAF1 protein function cause altered neuronal activity and measurable changes in target gene expression. Specific Aim 1 will determine the effects of newly identified DEAF1 mutations on DEAF1 protein function and also assess dominant negative activity of the DEAF1 mutant proteins using both in vitro and ex vivo approaches. Specific Aim 2 will evaluate the effects of diminished DEAF1 function on neuronal activity/plasticity and target gene expression. Mice with conditional knockout of Deaf1 in the brain show anxiety-like behavior and impaired contextual memory in fear-conditioning tests and will be used to establish a role of DEAF1 in regulating neuronal activity. DEAF1 gene targets related to neuronal and cognitive function that are differentially expressed in the hippocampus of these mice will be assessed. The proposed studies will describe the functional effects of identified human de novo DEAF1 mutations and the consequences of reduced DEAF1 activity in the central nervous system. We expect that the results of these studies will also provide evidence into the causal relationship between dysregulated DEAF1 function and intellectual disability.

描述（由申请人提供）：智力障碍的特点是认知能力低于平均水平，导致功能和适应行为显着缺陷。大多数不明原因的智力障碍病例是由新的基因突变引起的。发现了四名智力障碍患者。在转录因子变形表皮自动调节因子-1 (DEAF1) 的 DNA 结合域内具有杂合从头点突变，并且由此产生的氨基酸取代被证明会对DEAF1 蛋白功能。最近从智力障碍和自闭症谱系障碍患者的一组未识别样本中发现了另外四种 DEAF1 变体，这些突变也发生在 DNA 结合域内，但这些突变对 DEAF1 功能的影响是有限的。由于 DEAF1 单倍体充足性尚未被报道会导致智力障碍，这表明突变型 DEAF1 蛋白可能对野生型 DEAF1 功能产生显着的负面影响。该提议的假设是，DEAF1 基因的突变会削弱 DEAF1 蛋白的功能，导致神经活动发生改变，并且特定目标 1 的可测量变化将决定新发现的 DEAF1 的影响。突变对 DEAF1 蛋白功能的影响，并使用体外和离体方法评估 DEAF1 突变蛋白的显性负活性。具体目标 2 将评估 DEAF1 功能减弱对神经元活性/可塑性和靶基因的影响。大脑中条件性敲除 Deaf1 的小鼠在恐惧调节测试中表现出焦虑样行为和情境记忆受损，并将用于确定 DEAF1 在调节与神经元和认知功能相关的神经元活动中的作用。我们将评估这些小鼠海马体中的差异表达，所提出的研究将描述已识别的人类 DEAF1 突变的功能影响以及中枢神经系统中 DEAF1 活性降低的后果。这些研究还将为 DEAF1 功能失调与智力障碍之间的因果关系提供证据。