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Role of BDNF in dendritic pathologies caused by Rett-associated MeCP2 mutations

BDNF 在 Rett 相关 MeCP2 突变引起的树突状病理中的作用

基本信息

批准号：
7192018
负责人：
Lucas D Pozzo-Miller
金额：
$ 25.46万
依托单位：
UNIVERSITY OF ALABAMA AT BIRMINGHAM
依托单位国家：
美国
项目类别：
财政年份：
2007
资助国家：
美国
起止时间：
2007-04-01 至 2009-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7192018
关键词：
Affect Animal Model Animals Antibodies Arts Biolistics Birth Brain Brain-Derived Neurotrophic Factor Cell model Cells Chemicals Chromosome Pairing Chromosome abnormality Clinical Code Cues DNA-Binding Proteins Defect Dendrites Dendritic Spines Development Disease Exhibits Female Gene Targeting Gene Transfer Generations Genes Genetic Goals Hippocampus (Brain)Homeostasis Image Immunoglobulin G Impaired cognition Impairment Intervention Knockout Mice Lasers Learning Length Link Maintenance Measures Mediating Memory impairment Mental Retardation Messenger RNA Methyl-CpG-Binding Protein 2 Microscopy Mutation NGFR Protein Neurodevelopmental Disability Neurodevelopmental Disorder Neurons Pathology Patients Phenotype Process Property Research Personnel Rett Syndrome Reverse Transcriptase Polymerase Chain Reaction Role Scanning Severities Signal Transduction Slice Small Interfering RNA Structure Synapses Synaptic plasticity Syndrome Testing Therapeutic Intervention Thinking Time Time Study Transcriptional Regulation Transfection Transgenic Organisms Vertebral column Work base cell motility clinically relevant density developmental disease hippocampal pyramidal neuron insight interest mutant nervous system disorder neuronal survival neuropathology novel particle patch clamp physical insult postnatal postsynaptic presynaptic programs receptor research study

项目摘要

DESCRIPTION (provided by applicant): The neuropathology of mental retardation is thought to be associated with deficits in synaptic structure and function. Our goal is to make contributions into understanding the formation and maintenance of dendritic spines and of postsynaptic Ca2+ homeostasis in neurons expressing Rett Syndrome-associated mutations in MECP2. Rett syndrome (RTT) is an X- linked developmental disorder and the leading cause of mental retardation in females. Mutations in the transcriptional represser MECP2 have been identified in >90% of RTT cases. One of the target genes of MeCP2 is bdnf, brain-derived neurotrophic factor. Considering that BDNF has recently emerged as a potent modulator of activity-dependent synaptic development and plasticity in the postnatal brain, including fundamental neuronal properties such as dendritic spine density and form and neuronal Ca2+ signaling, we hypothesize that a deregulation of BDNF signaling may underlie the dendritic pathologies observed in RTT. The specific hypothesis to be tested is twofold: 1) RTT-associated MECP2 mutations cause dendritic spine loss leading to Impaired dendritic Ca2+ signaling in hippocampal pyramidal neurons through reduced BDNF signaling; 2) impaired dendritic structure in MECP2 mutant neurons can be reverted by BDNF treatment. The consequences of mutant MECP2 expression will be evaluated in neurons maintained in organotypic slice cultures and transfected by particle-mediated gene-transfer. The biolistic gene-transfer approach provides a more flexible way to introduce different mutant forms of MECP2 compared to the generation of transgenic or knockout mice, in addition to allow the co-transfection of other cDNAs or knockdown siRNA constructs of interest. Thus, it represents a novel cellular model of RTT. Transfected neurons will be studied by laser-scanning confocal and time-lapse multiphoton microscopy, as well as by simultaneous Ca2+ imaging and whole-cell intracellular recordings. This combination of state-of-the-art approaches has never been used to investigate MECP2 function, or applied to animal models of RTT. We expect the proposed studies to provide novel insights into the consequences of mutant MECP2 expression in hippocampal neurons in a cellular model of RTT.

描述(申请人提供)：智力低下的神经病理学被认为与突触结构和功能的缺陷有关。我们的目标是为理解树突棘的形成和维持以及表达Rett综合征相关MECP2突变的神经元的突触后钙稳态做出贡献。Rett综合征(RTT)是一种X连锁的发育障碍，是女性智力低下的主要原因。转录抑制因子MECP2的突变在90%的RTT病例中被发现。脑源性神经营养因子是MeCP2的靶基因之一。考虑到BDNF最近已成为出生后脑内活性依赖的突触发育和可塑性的有效调节器，包括树突棘密度和形状以及神经元钙信号等基本神经元特性，我们假设BDNF信号的解除调控可能是RTT中观察到的树突状病理的基础。需要检验的具体假设有两个：1)RTT相关的MECP2突变导致树突棘丢失，通过减少BDNF信号导致海马锥体神经元树突状钙信号受损；2)MECP2突变神经元中受损的树突状结构可以被BDNF治疗恢复。突变的MECP2表达的后果将在器官型切片培养中保持的神经元中进行评估，并通过颗粒介导的基因转移进行转移。与转基因或基因敲除小鼠相比，生物基因转移方法提供了一种更灵活的方法来引入不同突变形式的MECP2，此外还允许共转染其他感兴趣的cDNA或击倒siRNA构建体。因此，它代表了一种新的RTT细胞模型。转基因神经元将通过激光扫描共聚焦和延时多光子显微镜以及同步的钙成像和全细胞内记录进行研究。这种先进方法的结合从未被用于研究MECP2的功能，也从未被应用于RTT的动物模型。我们期待拟议的研究为RTT细胞模型中海马神经元MECP2突变表达的后果提供新的见解。