Neuronal development and synaptic plasticity in Cdc42 cenditional knockout mice

Cdc42 基因敲除小鼠的神经元发育和突触可塑性

• 批准号: 18300106
• 负责人: AIBA Atsu
• 金额: $ 10.53万
• 依托单位: Kobe University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 2006
• 资助国家: 日本
• 起止时间: 2006 至 2007
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-18300106/
• 关键词: Cdc42; knockout mice; Rho family; cerebral cortex; cerebellum; cdc42; Cre; Ella-Cre

In order to investigate the role of Rho family GTPase, Cdc42, in synapse formation and maintenance, synaptic plasticity, and leaning, we generated Cdc42 conditional knockout mice. We introduced loxP sequences into cdc42 allele by gene targeting in the embryonic stem cells. Then we generated flox-Cdc42 mice by using the mutant ES cells. By crossing flox-Cdc42 mice with two different neuron-specific Cre lines, we generated two distinct neuron-specific Cdc42 knockout lines.(1) Forebrain-specific Cdc42 knockout (FB-Cdc42 KO) miceTo avoid embryonic lethality, we have disrupted cdc42 gene via Cre-loxP recombination using Emx1-Cre mice. Emx1 promoter/enhancer induces Cre recombinase expression exclusively in the dorsal telencephalon as early as embryonic day (E) 10.5, tcogehereby eliminating Cdc42 expression in cortical projection neurons from the beginning of cerebral cortinesis. Western blot analysis of the protein prepared from FB-Cdc42 KO cerebral cortex showed that Cdc42 was knocked down in the KO cerebral cortex. We have found expanded cerebral cortex with abnormal layer formation in the FB-Cdc42 KO mice. Furthermore, the morphology of hippocampus was severely distorted in the FB-Cdc42 KO mice. These results suggested that Cdc42 controls the cell proliferation and differentiation of the neuron during cortical development.(2) Purkinje cell specific Cdc42 knockout (PC-Cdc42 KO) miceTo investigate the role of Cdc42 in cerebellar Purkinje cells, we have disrupted cdc42 gene using L7-Cre mice. PC-Cdc42 KO mice did not show ataxic gait. The histological analysis of the PC-Cdc42 KO cerebellum showed no apparent abnormality in morphology of the Purkinje cell dendrites.

为了研究Rho家族GTPase Cdc42在突触形成和维持、突触可塑性和学习中的作用，我们制造了Cdc42条件敲除小鼠。将loxP序列导入胚胎干细胞cdc42等位基因中。然后我们利用突变的ES细胞产生了flox-Cdc42小鼠。通过将flox-Cdc42小鼠与两种不同的神经元特异性Cre系杂交，我们产生了两种不同的神经元特异性Cdc42敲除系。(1)前脑特异性Cdc42敲除（FB-Cdc42 KO）小鼠为了避免胚胎致死，我们利用Emx1-Cre小鼠通过Cre-loxP重组来破坏Cdc42基因。Emx1启动子/增强子早在胚胎日(E) 10.5就诱导Cre重组酶仅在背端脑表达，从而从大脑皮质形成开始就消除了皮层投射神经元中Cdc42的表达。用Western blot方法检测KO大脑皮层中制备的FB-Cdc42蛋白，发现KO大脑皮层中Cdc42蛋白被敲低。我们发现FB-Cdc42 KO小鼠的大脑皮层扩大并形成异常层。此外，FB-Cdc42 KO小鼠海马形态严重扭曲。这些结果表明，在皮层发育过程中，Cdc42控制着神经元的细胞增殖和分化。(2)浦肯野细胞特异性Cdc42敲除（PC-Cdc42 KO）小鼠为了研究Cdc42在小脑浦肯野细胞中的作用，我们使用L7-Cre小鼠破坏Cdc42基因。PC-Cdc42 KO小鼠未表现出共济失调步态。PC-Cdc42 KO小脑的组织学分析显示浦肯野细胞树突形态未见明显异常。

项目成果

Semaphorin 3F confines ventral tangential migration of lateral olfactory tract neurons onto telencephalon surface

Semaphorin 3F 限制侧嗅束神经元向端脑表面的腹侧切向迁移

• 发表时间: 2008
• 期刊: J. Neurosci 28-17
• 作者: Ito;K.;Kawasaki;T.;Takashima;S.;Matsuda;I.;Aiba.;A.;Hirata;T
• 通讯作者: T

The M-Ras-RA-GEF-2-Rapl pathway mediates tumor necrosis factor-a-dependent regulation of integrin activation in splenocytes

M-Ras-RA-GEF-2-Rapl 通路介导脾细胞中肿瘤坏死因子 a 依赖性整合素激活调节

• 发表时间: 2007
• 期刊: Mol. Biol. Cell 18-8
• 作者: Yoshikawa;Y.;Satoh;T.;Tamura;T.;Wei;P.;Bilasy;S. E.;Edamatsu;H.;Aiba.;A.;Katagiri;K.;Kinashi;T.;Nakao;K.;Kataoka;T
• 通讯作者: T

mGluR1シグナルの遺伝学的解析及びプロテオミクス解析

mGluR1 信号的遗传和蛋白质组分析

• 发表时间: 2007
• 作者: Kassai;et.al.;饗場篤
• 通讯作者: 饗場篤

mGluR1 is essential for motor coordination in the adult cerebellum.

mGluR1 对于成人小脑的运动协调至关重要。

• 发表时间: 2007
• 作者: Aiba;A.;Nakao;H.;Nakao;K.;Kano;M.
• 通讯作者: M.

protein-independent neuromodulatory action of adenosine on metabotropic glutamate signaling in mouse cerebellar Purkinje Cells

腺苷对小鼠小脑浦肯野细胞代谢型谷氨酸信号传导的蛋白质独立神经调节作用

• 发表时间: 2007
• 期刊: J. Physiol 581-2
• 作者: Tabata;T.;Kawakami;D.;Hashimoto;K.;Kassai;H.;Yoshida;T.;Hashimotodani;Y.;Fredholm;B. B.;Seikno;Y.;Aiba;A.;Kano;M. G
• 通讯作者: M. G