Regulation of synaptic actin reorganization by signal transmission with drebrin family

通过与drebrin家族的信号传递调节突触肌动蛋白重组

• 批准号: 12480236
• 负责人: SHIRAO Tomoaki
• 金额: $ 9.54万
• 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: 2000
• 资助国家: 日本
• 起止时间: 2000 至 2001
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-12480236/
• 关键词: drebrin; dendritic spine; actin; synaptic plasticity; signal transmission; neuronal culture; cytoskeleton; SH3P7; シナプス; アクチン結合蛋白; 培養; 神経細胞

Drebrins are actin-binding proteins, which expression is closely related to spine morphology. Therefore we hypothesized that drebrin-family proteins play adaptor proteins of postsynaptic signal transmission from membrane receptor to actin cytoskeleton.In the previous study, we established the primary culture of cortical neurons, in which neurons can develop and the dendritic spine seemed to be fully maturated. Using this culture systems, we inhibit the drebrin expression wit antisense-oligonucleotides, and demonstrated that activity-dependent accumulation of synaptic functional proteins were changed.Next, we hypothesized that other drebrin-binding proteins than actin play an important role in postsynaptic signal transmission in the neuron. We performed the yeast two hybrid system for cloning the binding proteins to drebrin-family proteins (drebrin E, drebrin A, and SH3P7).One of the clone, designated as DRAP1, encodes full length of a coda of a novel drebrin-binding protein. Then we performed its biochemical characterization and found its N-terminal half actually binds to the N-terminal region of a drebrin molecule. Next, using polypeptide of DRAP1 and DRAP1 fragment expressed by gene engineering technique, we raise polyclonal antiserum to DRAP1.Further, we constructed expression vector which encode the GFP-DRAP1 fused protein. When we transformed fibroblasts with the expression vector, we observed GFP fluorescence in their nuclei.

DREBRINS是肌动蛋白结合蛋白，表达与脊柱形态密切相关。因此，我们假设Drebrin-家庭蛋白在从膜受体到肌动蛋白细胞骨架的突触后信号传播中发挥衔接蛋白。 。使用这种培养系统，我们抑制了反义 - 寡核苷酸的DREBRIN表达，并证明改变了突触功能蛋白的活动依赖性依赖性的积累。隔离，我们假设其他DREBRIN结合蛋白比肌动蛋白在肌动蛋白中比肌动蛋白在突触后信号传播中起着重要作用神经元。我们执行了酵母两种混合系统，用于将结合蛋白克隆到DREBRIN-家族蛋白（Drebrin E，Drebrin A和SH3P7）。一种被称为DARAP1的克隆的一种，编码了新型Drebrin结合蛋白的编码。然后，我们进行了其生化表征，发现其N末端一半实际上与Drebrin分子的N末端区域结合。接下来，使用基因工程技术表达的drap1和drap1片段的多肽，我们将多克隆抗血清提高到drap1。当我们用表达载体转化成纤维细胞时，我们观察到其核中的GFP荧光。

项目成果

Y. Tomidokoro: "Brain Aβ amyloidosis in APPsw mice induces accumulation of presenilin-I and tau"Journal of Pathology. 193. 500-506 (2001)

Y. Tomidokoro：“APPsw 小鼠的脑 Aβ 淀粉样变性诱导早老素-I 和 tau 蛋白的积累”《病理学杂志》193. 500-506 (2001)。

手塚 美佳: "Cellular and molecular changes during the process of repair of spinal cord injury in infant rats"Neurotrauma Research. 12. 57-60 (2000)

Mika Tezuka：“幼年大鼠脊髓损伤修复过程中的细胞和分子变化”神经创伤研究。12. 57-60 (2000)。

S. Tanaka, Y. Sekino, T. Shirao: "NT-3 inhibits cerebellar granule cell migration in vitro"Neurosci.. 97. 727-734 (2000)

S. Tanaka、Y. Sekino、T. Shirao：“NT-3 在体外抑制小脑颗粒细胞迁移”Neurosci.. 97. 727-734 (2000)

金明鎬: "A Novel Brain-Specific Mouse Drebrin : cDNA Cloning, Chromosomal Mapping, Genomic Structure, Expression, and Functional Characterization"Genomics. (2002)

Kim Myung-ho：“新型脑特异性小鼠 Drebrin：cDNA 克隆、染色体作图、基因组结构、表达和功能表征”基因组学 (2002)。

M Ikeda, M Segara, Y Sekino, T Shirao, K Honda, T Yoshioka, CN Allen, S Inoue: "Complete sleep deprivation and Blocking of A1 adenosine receptor-mediated inhibition of intracellular calcium signaling in the rat ventromedial preoptic nucleus by sulphydryl

M Ikeda、M Segara、Y Sekino、T Shirao、K Honda、T Yoshioka、CN Allen、S Inoue：“通过巯基完全剥夺睡眠并阻断 A1 腺苷受体介导的大鼠腹内侧视前核细胞内钙信号传导抑制