Role of actin cytoskeleton in the axonal growthcone during brain development

肌动蛋白细胞骨架在大脑发育过程中轴突生长锥中的作用

• 批准号: 16300117
• 负责人: SHIRAO Tomoaki
• 金额: $ 9.47万
• 依托单位: Gunma University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 2004
• 资助国家: 日本
• 起止时间: 2004 至 2006
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-16300117/
• 关键词: actin; growthcone; hippocampal neuronal culture; immunofluorescence staining; actin-binding proteins; axonal growth; microtubule; 微小管

During the neuronal network formation, the inhibition of axonal growthcone morphogenesis and its function make it impossible for the brain function to develop normally. In this study, in order to elucidate the regulatory mechanism of actin cytoskeleton, which is directly involved in the morphogenesis and function of axonal growthcones, we focused in the actin filament and analyzed the change in the protein network of actin-binding proteins which regulate the physicochemical and biochemical characters using primary cultured hippocampal neurons. We first analyzed the distribution of drebrin in the axonal growthcone of cultured hippocampal neurons at various developmental stages using immunocytochemistry. In the neuron of stage 2, drebrin was localized at transitional area of growthcones. The immunocytochemistry also showed that Neurabin 1 showed similar localization of in the growthcone to drebrin. We further analyzed localization of various actin-related proteins in the growthcones at stage 2. Microtubules did not invade into the transitional area. In order to discriminate drebrin E from drebrin A, we developed drebrin A-specific antibody. This antibody clearly showed that growthcone only has drebrin E. This antibody further showed that migrating neurons in the adult brain only express drebrin E. We developed the new identification method of migrating neurons in the adult brain. Using this method, we discovered migrating neurons in the piriform cortex in adult rats. We finally showed that drebrin knockdown by RNAi resulted the inhibition of axonal growth.

在神经元网络形成过程中，轴突生长锥的形态发生和功能受到抑制，使脑功能不能正常发育。本研究以原代培养的海马神经元为研究对象，以肌动蛋白微丝为研究对象，分析了肌动蛋白结合蛋白网络的变化，探讨了肌动蛋白骨架对轴突生长锥形态发生和功能的调控机制。我们首先用免疫细胞化学的方法分析了不同发育阶段培养的海马神经元轴突生长锥中dreglycoprotein的分布。在第2阶段的神经元中，神经元的轴突位于生长锥的过渡区。免疫细胞化学结果显示Neurabin 1在生长锥中的定位与dreabin相似。我们进一步分析了各种肌动蛋白相关蛋白在第2阶段生长锥的定位。移行区未见微管侵入。为了区分dreA和dreE，我们研制了dreA特异性抗体。该抗体清楚地表明生长锥仅具有dreplase E。该抗体进一步表明，成年脑中的迁移神经元仅表达dreplase E。我们发展了一种新的成人脑内迁移神经元的鉴定方法。利用这种方法，我们发现了成年大鼠梨状皮质中的迁移神经元。我们最后发现，通过RNAi敲低dreglycoprotein导致轴突生长抑制。

项目成果

AMPA receptor downscaling at the onset of Alzheimer's pathology in double knock-in mice

双敲入小鼠阿尔茨海默病病理发生时 AMPA 受体的下调

• 发表时间: 2006
• 期刊: Proc. Natl. Acad Sci.(USA) 103
• 作者: Chang EH;Savage MJ;Flood DG;Thomas JM;Levy RB;Mahadomrongkul;V.;Shirao;T.;Aoki;C.;Huerta;P.T.
• 通讯作者: P.T.

興奮性シナプスのアクチン結合タンパク-その動態と機能-

兴奋性突触的肌动蛋白结合蛋白 - 它们的动力学和功能 -

• 发表时间: 2006
• 期刊: 蛋白質・核酸・酵素 51
• 作者: Sekino;Y.;関野 祐子
• 通讯作者: 関野 祐子

Synaptic dysfunction and disruption of the postsynaptic drebrin-actin complex: the study of neurological disorders accompanied by cognitive deficits

• 发表时间: 2007
• 期刊: Neuropharmacology
• 影响因子: 4.7
• 作者: N. Kojima;T. Shirao

Antisense knockdown of drebrin A, a dendritic spine protein, causes stronger preference, impaired pre-pulse inhibition, and an increased sensitivity to psychostimulant

• DOI: 10.1016/j.neures.2004.02.014
• 发表时间: 2004-06-01
• 期刊: NEUROSCIENCE RESEARCH
• 影响因子: 2.9
• 作者: Kobayashi, R;Sekino, Y;Saji, M
• 通讯作者: Saji, M

Downregulation of drebrin A expression suppresses synaptic targeting of NMDA receptors in developing hippocampal neurons

drebrin A 表达下调抑制发育中海马神经元中 NMDA 受体的突触靶向

• 发表时间: 2006
• 期刊: J. Neuochem 97(sl)
• 作者: Takahashi;H.;Mizui T.;Shirao;T.
• 通讯作者: T.