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Deciphering high function autism using mice with human de novo ANK2 mutations

使用携带人类 ANK2 新突变的小鼠解读高功能自闭症

基本信息

批准号：
9429362
负责人：
G Vann Bennett
金额：
$ 23.88万
依托单位：
DUKE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2017
资助国家：
美国
起止时间：
2017-09-18 至 2019-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9429362
关键词：
ANK2 gene Ankyrins Arrhythmia Autistic Disorder Axon Behavior Behavioral Binding Brain Brain region Cell Adhesion Molecules Cellular Assay Cellular biology Cherry - dietary Cognition Collaborations Computer Assisted Core Facility Data Development Developmental Delay Disorders Diffusion Magnetic Resonance Imaging Electrophysiology (science)Exhibits Exocytosis Exons Frameshift Mutation Genes Goals Growth Cones Human Image Image Analysis Immunofluorescence Immunologic Injection of therapeutic agent Intellectual functioning disability Label Length Metabolic syndrome Missense Mutation Modeling Molecular Motor Mouse Strains Mus Mutant Strains Mice Mutation Nervous system structure Neural Cell Adhesion Molecule L1 Neurons Neurosciences Neurosecretory Systems Nonsense Mutation Phenotype Prosencephalon Psychology Reproducibility Research Research Personnel Resources Role Seizures Sensory Signal Transduction Social Behavior Symptoms Synapses Three-dimensional analysis Variant Yin anxiety-like behavior autism spectrum disorder axon growth base brain morphology cell type conditional mutant human data in vivo insight knock-down mouse model mutant myelination polypeptide postnatal premature relating to nervous system synaptogenesis

项目摘要

The long-term goal of this research is to resolve molecular and cellular substrates for high-functioning autism. We will develop mouse models for human de novo mutations in ANK2, one of the few genes associated with autism in the absence of intellectual disability, developmental delay, or seizures. ANK2 encodes 220 kDa ankyrin-B (ankB) which is widely expressed, and 440 kDa ankB which is found only in neurons and is localized to unmyelinated axons and growth cones. The distribution of human de novo mutations suggests that mutation of 440 kDa ankB is sufficient to cause high function autism. We have generated mice modeling a human de novo ANK2 frameshift mutation that selectively depletes 440 kDa ankB. We will explore the hypothesis that mutation/deficiency of 440 kDa ankB selectively perturbs both local and long-range neural connectivity through altered axonal function as follows. 1. Resolve the molecular basis for increased axon growth/branching observed in 440 kDa ankB mutant neurons, and establish a cellular assay to evaluate human 440 kDa ankB variants. Neurons cultured from mice lacking 440 kDa ankB exhibit increased axon length and branching. We will explore the roles in this phenomenon of cytoskeletal dynamics and exocytosis. We also will develop a rescue strategy to evaluate functional consequences of human variants of 440 kDa ankB on axon growth/branching. 2. Determine consequences of 440 kDa ankB mutation for establishment of local and long-range neural connectivity during postnatal brain development. 440 kDa ankB-deficient mice exhibit increased cortical synaptogenesis as well as loss of interaction with the cell adhesion molecule L1CAM. Since L1CAM requires ankyrin-binding activity for long distance axon pathfinding, we hypothesize that 440 kDa ankB-deficient neurons will exhibit abnormal axon targeting. We will evaluate local and long-range targeting of axons of during development in 440 kDa ankB-deficient mice using a) diffusion tensor imaging (DTI) b) imaging of axons of cortical neurons sparsely labeled by stereotactic injection of AAV GFP/Cherry constructs; c) analysis of excitatory and inhibitory forebrain synapses during development using immunofluorescence and electrophysiology. 3. Determine a reproducible behavioral phenotype for 440 kDa ankB mutant mice. In collaboration with Drs. William Wetsel (Director of the Duke Mouse Behavioral and Neuroendocrine Analysis Core Facility) and Henry Yin (Duke Department of Psychology and Neuroscience) and co-investigators on this proposal, we will evaluate social behavior, cognition, sensory-motor gating, and repetitive/anxiety-like behavior. In addition, in collaboration with Henry Yin, we will use computer-assisted analysis of 3D imaging to develop an unbiased reproducible behavior profile. These studies of cellular and in vivo consequences of de novo ANK2 mutation promise to provide the first mechanistic insights into high functioning forms of autism.

这项研究的长期目标是解决高功能自闭症的分子和细胞基础。