Production and analyses of mice deficient in molecules involved in cell polarization

细胞极化相关分子缺陷小鼠的生产和分析

• 批准号: 14370003
• 负责人: HARADA Akihiro
• 金额: $ 8.38万
• 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: 2002
• 资助国家: 日本
• 起止时间: 2002 至 2004
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-14370003/
• 关键词: polarized transport; cell polarity; epithelial cell; neuron; Cre-loxP; gene targeting; rab proteins; SNARE proteins; TGN; rab; SNARE; ノックアウトマウス; Cre; loxP

Cell polarity is crutial for function of various types of cells. For cell polarity, polarized transport within cells are thought to be essential. A number of proteins (e.g, rab proteins and SNARE proteins) have been shown to be involved in polarized transport. However, their roles in the development and function of tissues and whole body of animals are largely unknown. Therefore, to uncover their roles in the development and function in the animals, we are making knockout mice for each of these proteins. As it is possible that these molecules are involved in embryonic development, simply knocking out of these genes may lead to embryonic lethality at very early stages of development. To circumvent this problem, we are making conditional knockout mice using Cre-loxP system.During the term of grant, we have completed making conditional knockout mice of these proteins and are now analyzing their phenotypes. In addition, we analyzed AP3B knockout mice under electron microscopy and showed that the diameter of their synaptic vesicles became enlarged, which may lead to the neurological phenotype (convulsion) of this knockout mice. We also looked at the polarization process of neurons in vitro and observed that (1)axons were able to be formed from dendrites (2)axonogenesis is delayed in inhibitory neurons compared to excitatory neurons.

细胞极性对于各种类型细胞的功能都是残酷的。对于细胞极性，细胞内的极化转运被认为是必不可少的。许多蛋白质（例如，Rab蛋白和SNARE蛋白）已显示出参与极化转运。然而，它们在组织和整个动物身体的发展和功能中的作用在很大程度上是未知的。因此，为了揭示它们在动物的发育和功能中的作用，我们正在为每种蛋白质制作敲除小鼠。由于这些分子可能参与胚胎发育，因此仅仅从这些基因中删除可能会导致在发育的早期阶段就会导致胚胎致死性。为了解决这个问题，我们正在使用Cre-loxp系统制造有条件的基因敲除小鼠。在授予的术语中，我们已经完成了这些蛋白质的有条件敲除小鼠，现在正在分析其表型。此外，我们在电子显微镜下分析了AP3B敲除小鼠，并表明它们的突触囊泡的直径增大，这可能导致该基因敲除小鼠的神经系统型（抽搐）。我们还研究了体外神经元的极化过程，并观察到（1）轴突能够由树突形成（2）与兴奋性神经元相比，抑制性神经元的轴突发生延迟。

项目成果

Reversal of neuronal polarity characterized by conversion of dendrites into axons in neonatal rat cortical neurons in vitro.

体外新生大鼠皮质神经元中神经元极性的逆转以树突转化为轴突为特征。

• 发表时间: 2002
• 期刊: Neuroscience 110
• 作者: Kensuke Hayashi

MAP2 is required for dendrite elongation, PKA anchoring in dendrites, and proper PKA signal transduction.

• DOI: 10.1083/jcb.200110134
• 发表时间: 2002-08-05
• 期刊: The Journal of cell biology
• 作者: Harada A;Teng J;Takei Y;Oguchi K;Hirokawa N
• 通讯作者: Hirokawa N

Defective function of GABA-containing synaptic vesicles in mice lacking the AP-3B clathrin adaptor.

• DOI: 10.1083/jcb.200405032
• 发表时间: 2004-10-25
• 期刊: JOURNAL OF CELL BIOLOGY
• 影响因子: 7.8
• 作者: Nakatsu, Fubito;Okada, Motohiro;Mori, Fumiaki;Kumazawa, Noriko;Iwasa, Hiroto;Zhu, Gang;Kasagi, Yasufumi;Kamiya, Haruyuki;Harada, Akihiro;Nishimura, Kazuhiro;Takeuchi, Arata;Miyazaki, Taisuke;Watanabe, Masahiko;Yuasa, Shigeki;Manabe, Toshiya;Wakabayashi, Koichi;Kaneko, Sunao;Saito, Takashi;Ohno, Hiroshi
• 通讯作者: Ohno, Hiroshi

Kensuke Hayashi: "Reversal of neuronal polarity characterized by conversion of dendrites into axons in neonatal rat cortical neurons in vitro"Neuroscience. 110・1. 7-17 (2002)

Kensuke Hayashi：“体外新生大鼠皮质神经元中以树突转化为轴突为特征的神经元极性逆转”神经科学 110・1（2002）。

Kensuke Hayashi: "Inhibitory neurons from fetal rat cerebral cortex exert delayed axon formation and active migration in vitro."Journal of Cell Science. 116・21. 4419-4428 (2003)

Kensuke Hayashi：“来自胎鼠大脑皮层的抑制性神经元在体外发挥延迟的轴突形成和主动迁移。”细胞科学杂志 116・21 4419-4428（2003）。