Disabled-1可变性剪接在调控Reelin信号通路和神经元迁移中的分子研究

项目名称： Disabled-1可变性剪接在调控Reelin信号通路和神经元迁移中的分子研究

项目编号： No.31300905

项目类型： 青年科学基金项目

立项/批准年度： 2014

项目学科： 生物科学

项目作者： 高志华

作者单位： 浙江大学

项目金额： 28万元

中文摘要： Reelin是调节神经元迁移的关键分子，异常与自闭症、抑郁症等密切相关。连接蛋白Dab1是Reelin信号通路的核心，经Reelin刺激后可发生酪氨酸磷酸化，并激活下游效应分子。目前研究仅局限于单个Dab1分子，然而，Dab1协调细胞迁移的机制至今未明。我们的前期研究发现Dab1受剪接调控能产生多达11种剪接体，颠覆了传统的单一Dab1论。进一步研究表明可变性剪接能在时空上精确控制Dab1中酪氨酸位点的数目和磷酸化状态，提示多个Dab1剪接体能共同参与Reelin信号通路并协调神经元的迁移。本项目在前期研究基础上，采用胚胎电转染的方法，在体引入ASOs阻断内源剪接，表达外源剪接体，结合实时荧光显微成像和生化手段，深入研究Dab1剪接在调控神经元迁移中的作用。本项目首次从剪接角度研究细胞迁移，将为阐明神经元迁移的分子机制提供新方向，为揭示Reelin相关精神疾病的发病机制提供新的理论依据。

中文关键词： 神经元迁移；Reelin；Disabled-1；RNA 剪接；磷酸化

英文摘要： Reelin plays an instrumental role in regulating neuronal migration. Aberrant Reelin signaling is associated with multiple psychiatric disorders including autism and schizophrenia. The intracellular adaptor protein Disabled-1 (Dab1) is key to Reelin signaling. Reelin stimulation induces Dab1 tyrosine phosphorylation, which then recruits and activates downstream cascades. To date, worldwide studies have primarily focused on the role of a single Dab1 protein in neuronal migration; yet, mechanisms underlying the orchestration of neuronal migration remain unclear. Contrary to the classic view, we have previously discovered that Dab1 is alternatively spliced during brain development, generating up to 11 Dab1 isoforms. Further studies have revealed that alternative splicing precisely controls the number and phosphorylation status of tyrosine sites within Dab1 isoforms in a spatio-temporal manner. Our data suggest that multiple Dab1 isoforms co-ordinate the downstream signaling and dynamically orchestrate neuronal migration. In this grant proposal, we plan to further determine the role of Dab1 alternative splicing in neuronal migration in vivo, using antisense oligonucleotides (ASOs) to specifically block the alternative splicing of Dab1, in utero electroporation to ectopically express Dab1 isoforms, in combination with

英文关键词： neuronal migration；Reelin；Disabled-1；RNA splicing；phosphorylation