Dynamics of Synaptic Proteins in C.elegans

线虫突触蛋白的动力学

• 批准号: 15500208
• 负责人: KUROYANAGI Hidehito
• 金额: $ 2.37万
• 依托单位: Tokyo Medical & Dental University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2003
• 资助国家: 日本
• 起止时间: 2003 至 2004
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-15500208/
• 关键词: C.elegans; visualization; fluorescent protein; glutamate receptor; synapse; 経時観察

Long-term visualization of synaptic proteins and spine morphology in mammalian neurons revealed continual turnover of a fraction of synapse population in vitro and in vivo. Furthermore, recent analysis of mammalian synaptic proteins showed rapid alterations of protein compositions in synapses and their regulation by neuronal activity. Significance of continual alterations of synaptic structure and molecular compositions should be evaluated in a system where direct correlational analyses between changes of circuit properties and resulting behavioral alterations are possible. In C.elegans, full descriptions of the circuitry of all 302 neurons are available and specific sets of neurons and genes that control the sensory-motor coordination have been identified. Therefore, long-term visualization of synaptic proteins essential in the proper synaptic functions between neurons involved in the sensory-motor coordination would be an ideal model system to test the behavioral significance of syna … More pse remodeling.To this end, we focused on the glutamatergic synapses between polymodal sensory neuron ASH and interneurons of the locomotory control circuit (command neurons) of C.elegans. The glutamatergic transmission at ASH-command neuron synapses is essential for response to nose-touch. To visualize postsynaptic component, we utilized a fusion protein between ionotropic glutamate receptor GLR-1 and green fluorescent protein (GLR-1::GFP) and expressed this reporter gene either by an authentic glr-1 promoter or nmr-1 promoter that support expression of GLR-1 in a subset of interneurons. Images were obtained by a two-photon laser scanning microscope. Transgenic worms expressing GLR-1-GFP under the control of glr-1 promoter showed fluorescent clusters in both the nerve ring and the ventral nerve cord. Time-lapse imaging of short- and long-term remodeling of GLR-1::GFP clusters in the ventral nerve cord revealed both short-term structural stability and development-dependent remodeling. In transgenic worms expressing GLR-1::GFP under the control of nmr-1 promoter, a smaller number of fluorescent clusters were visualized within the nerve ring. Anatomical location and double labeling of ASH neurons with longer-wavelength fluorescent probes suggested that some of the GLR-1::GFP clusters corresponded to the synaptic junctions between ASH and command neurons. Less

在哺乳动物神经元突触蛋白和棘形态的长期可视化揭示了在体外和体内突触群体的一部分的持续营业额。此外，最近对哺乳动物突触蛋白的分析表明，突触中的蛋白质组成及其受神经元活动调节的快速变化。突触结构和分子组成的连续变化的意义应在一个系统中进行评估，其中电路特性的变化和由此产生的行为改变之间的直接相关分析是可能的。在秀丽隐杆线虫中，所有302个神经元的电路的完整描述是可用的，并且已经确定了控制感觉-运动协调的特定神经元和基因组。因此，对参与感觉-运动协调的神经元之间的正确突触功能所必需的突触蛋白的长期可视化将是测试突触蛋白的行为意义的理想模型系统。 ...更多信息 神经重塑。为此，我们专注于多模态感觉神经元ASH和运动控制回路（命令神经元）的中间神经元的秀丽隐杆线虫之间的神经元突触。在对鼻触的反应中，在ASH-命令神经元突触处的突触能传递是必不可少的。为了可视化突触后成分，我们利用离子型谷氨酸受体GLR-1和绿色荧光蛋白（GLR-1：：GFP）之间的融合蛋白，并通过支持GLR-1在中间神经元亚群中表达的真实glr-1启动子或nmr-1启动子表达该报告基因。通过双光子激光扫描显微镜获得图像。在glr-1启动子控制下表达GLR-1-GFP的转基因蠕虫在神经环和腹神经索中都显示荧光簇。对腹神经索中GLR-1：：GFP簇的短期和长期重塑的延时成像显示了短期结构稳定性和发育依赖性重塑。在nmr-1启动子控制下表达GLR-1：：GFP的转基因蠕虫中，在神经环内观察到较少数量的荧光簇。解剖位置和双标记的ASH神经元与较长波长的荧光探针表明，一些GLR-1：：GFP集群对应的ASH和命令神经元之间的突触连接。少

项目成果

Synchronized Formation and Remodeling of Postsynaptic Densities: Long-Term Visualization of Hippocampal Neurons Expressing Postsynaptic Density Proteins Tagged with Green Fluorescent Protein

• DOI: 10.1523/jneurosci.23-06-02170.2003
• 发表时间: 2003-03
• 期刊: The Journal of Neuroscience
• 作者: T. Ebihara;I. Kawabata;S. Usui;K. Sobue;S. Okabe

Impaired cell cycle control of neuronal precursor cells in the neocortical primordium of presenilin-1-deficient mice.

presenilin-1 缺陷小鼠新皮质原基中神经元前体细胞的细胞周期控制受损。

• 发表时间: 2002
• 期刊: J Neurosci Res. 70
• 作者: Yuasa S;Nakajima M;Aizawa H;Sahara N;Koizumi K;Sakai T;Usami M;Kobayashi S;Kuroyanagi H;Mori H;Koseki H;Shirasawa T.
• 通讯作者: Shirasawa T.

Electroporation-mediated gene transfer system applied to cultured CNS neurons

• DOI: 10.1097/00001756-200404290-00008
• 发表时间: 2004-04-29
• 期刊: NEUROREPORT
• 影响因子: 1.7
• 作者: Kawabata, I;Umeda, T;Okabe, S
• 通讯作者: Okabe, S

Bi-directional regulation of postsynaptic cortactin distribution by BDNF and NMDA receptor activity

• DOI: 10.1111/j.1460-9568.2005.04510.x
• 发表时间: 2005-12-01
• 期刊: EUROPEAN JOURNAL OF NEUROSCIENCE
• 影响因子: 3.4
• 作者: Iki, J;Inoue, A;Okabe, S
• 通讯作者: Okabe, S

Simultaneous observation of stably associated presynaptic varicosities and postsynaptic spines : morphological alterations of CA3-CA1 synapses in hippocampal slice cultures.

同时观察稳定相关的突触前静脉曲张和突触后棘：海马切片培养物中 CA3-CA1 突触的形态变化。

• 发表时间: 2005
• 期刊: Molecular and Cellular Neuroscience 28
• 作者: Umeda;T.;Ebihara;T.;S.Okabe
• 通讯作者: S.Okabe