Functional analysis of Cdk5 by using conditional KO of its activating subunit p35

通过条件性敲除其激活亚基p35对Cdk5进行功能分析

• 批准号: 21500356
• 负责人: OHSHIMA Toshio
• 金额: $ 2.91万
• 依托单位: Waseda University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2009
• 资助国家: 日本
• 起止时间: 2009 至 2011
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-21500356/
• 关键词: キナーゼ; リン酸化; コンディショナルKO

The purpose of this study is to analyze the function of neuron-specific serine/threonine kinase Cyclin-dependent kinase 5(Cdk5) by using conditional KO mice of its activating subunit p35 trough the behavioral analysis such as spatial learning and fear conditioning test, and electrophysiological analysis and biochemical analysis. In this study we generated p35-flox mice in which two loxP sequences were inserted in p35 gene allele. We generated inducible p35 conditional KO by crossing p35-flox and CreER mice in which Cre is activated by Tamoxifen(TM) and p35 gene was destructed after brain formed. We confirmed decrease of p35 protein by p. o. administration of 2 mg TM for three days. We also generated CA1-specific p35 conditional KO mice by crossing CaMKII-Cre mice, and confirmed decreased level of p35 protein in hippocampus. We observed decreased numbers of dendritic spines in the hippocampal CA1 pyramidal neurons in CA1-p35 cKO mice. Using these p35 conditional KO mice, we currently conduct behavioral analysis of p35 cKO mice.

本研究的目的是通过空间学习和恐惧条件反射测试等行为分析、电生理分析和生化分析，通过条件性敲除其激活亚基p35的小鼠，分析神经元特异性丝氨酸/苏氨酸激酶细胞周期蛋白依赖性激酶5（Cdk5）的功能。在这项研究中，我们生成了 p35-flox 小鼠，其中两个 loxP 序列插入到 p35 基因等位基因中。我们通过p35-flox和CreER小鼠杂交产生了诱导性p35条件KO，其中Cre被Tamoxifen(TM)激活，并且p35基因在大脑形成后被破坏。我们证实 p35 蛋白减少了 p。 o。给予 2 mg TM 三天。我们还通过CaMKII-Cre小鼠杂交产生了CA1特异性p35条件KO小鼠，并证实海马中p35蛋白水平降低。我们观察到 CA1-p35 cKO 小鼠海马 CA1 锥体神经元中树突棘数量减少。使用这些 p35 条件 KO 小鼠，我们目前对 p35 cKO 小鼠进行行为分析。

项目成果

Role of collapsin response mediator proteins for dendritic development in hippocampal CA1 pyramidal neurons

塌陷反应介导蛋白在海马 CA1 锥体神经元树突发育中的作用

• 发表时间: 2010
• 作者: Niisato E;Yamashita N;Nakamura F;Goshima Y;Ohshima T
• 通讯作者: Ohshima T

Hypomyelination Phenotype caused by impaired differentiation of oligodendrocytes in Emxl-cre mediated Cdk5 conditional knockout mice.

Emxl-cre 介导的 Cdk5 条件敲除小鼠中少突胶质细胞分化受损导致的低髓鞘化表型。

• 发表时间: 2011
• 期刊: Neurochemical Research
• 影响因子: 4.4
• 作者: Xe H;Takahashi S;Suzuki H;Hashikawa T;Kulkarni AB;Mikoshiba K;Ohshima T
• 通讯作者: Ohshima T

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Conditional deletion of neuronal cyclin-dependent kinase 5 in developing forebrain results in microglial activation and neurodegeneration.

• DOI: 10.2353/ajpath.2010.081158
• 发表时间: 2010
• 期刊: The American journal of pathology
• 作者: Satoru Takahashi;T. Ohshima;Motoyuki Hirasawa;T. Pareek;T. Bugge;Alexei Morozov;K. Fujieda;R. Brady;A. Kulkarni

Phosphorylation of AATYK1 by Cdk5 suppresses its tyrosine phosphorylation.

Cdk5 磷酸化 AATYK1 可抑制其酪氨酸磷酸化。

• 发表时间: 2010
• 期刊: PLoS ONE 5
• 作者: Tsutsumi;K.;Takano;T.;Endo;R.;Fukuda;M.;Ohshima;T.;Tomomura;M.;Hisanaga S.
• 通讯作者: Hisanaga S.