Measurement and Visualization of membrane potential by genetical manipul ati on.

通过基因操作测量和可视化膜电位。

• 批准号: 12557016
• 负责人: TAKUMI Toru
• 金额: $ 8.64万
• 依托单位: OSAKA BIOSCIENCE INSTITUTE(OBI) (2001)Kobe University (2000)
• 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-12557016/
• 关键词: membrane potential; neuroscience; physiology; Brain neuron; voltage-sensitive dyes; ニューロン

In neuroscience it is important to analyze not only a single neuron but also network communication among multiple neurons. It is not easy for biochemists or molecular biologists to use conventional electrophysiology when they discuss electrophysiological phenomenon in the nervous system. We plan to develop an untouched voltage-sensing probe and establish a novel assay system, without using conventional electrophysiology, to analyze the neuronal functions, combined with voltagesensitive dyes and the imaging system.We developed electrophysiological analyses and assay systems with use of voltage-sensitive dyes. We recorded long-term potentiation (LTP) in CAl and CA3 of mouse hippocampus and made a study of mechanisms of pharmacological modification. Further, we succeeded the recording of LTP in mouse hippocampal neurons using multi-electrodes.The system we develop may be possible to be mechanized or robotized automatically. It is thus expected that the system should be contribute to not only research in basic medicine but also industrialization such as the systematic screening of the compounds. The system using voltagesensitive dyes may enable us to analyze interaction among multi-neurons, in addition to study a single neuron.

在神经科学中，不仅要分析单个神经元，而且要分析多个神经元之间的网络通信。生物化学家或分子生物学家在讨论神经系统的电生理现象时，不容易使用传统的电生理学。我们计划开发一种非接触式电压传感探针，并建立一种新的检测系统，不使用传统的电生理学，结合电压敏感染料和成像系统来分析神经元的功能。我们开发了使用电压敏感染料的电生理分析和检测系统。我们记录了小鼠海马CAl和CA3的长时程增强（LTP），并对其药理作用机制进行了研究。此外，我们成功地利用多电极记录了小鼠海马神经元的LTP。我们开发的系统有可能实现机械化或自动化。因此，该系统不仅可以用于基础医学研究，还可以用于化合物的系统筛选等产业化。除了研究单个神经元外，使用电压敏感染料的系统还可以分析多个神经元之间的相互作用。

项目成果

H. Ohyama, H. Kajita, K. Omori, T. Takumi, N. Hiramoto, T. Iwasaka and H. Matsuda: "Inhibition of cardiac delayed rectifier K^+ currents by an antisense oligodeoxynucleotide against Isk (MinK) and over-expression of Isk mutant D77N in neonatal mouse heart

H. Ohyama、H. Kajita、K. Omori、T. Takumi、N. Hiramoto、T. Iwasaka 和 H. Matsuda：“针对 Isk (MinK) 和过度表达的反义寡脱氧核苷酸对心脏延迟整流 K^ 电流的抑制

S.Bouret, M.T.V.Chuoi-Mariot, V.Prevot, D.Croix, T.Takumi, S.Jegou, H.Vaudry, J.-C.Beauvilain, V.Mitchell: "Evidence that TGF-b may directly modulate POMC mRNA expression in the female rat arcuate nucleus"Endocrinology. 142. 4055-4065 (2001)

S.Bouret、M.T.V.Chuoi-Mariot、V.Prevot、D.Croix、T.Takumi、S.Jegou、H.Vaudry、J.-C.Beauvillain、V.Mitchell：“TGF-b 可能直接调节 POMC 的证据

H.Ohyama, H.Kajita, K.Omori, T.Takumi, N.Hiramoto, T.Iwasaka, H.Matsuda: "Inhibition of cardiac delayed rectifier K^+ currents by an antisense oligodeoxynucleotide against lsk (MinK) and over-expression of lsk mutant D77N in neonatal mouse hearts"Pflugers

H.Ohyama、H.Kajita、K.Omori、T.Takumi、N.Hiramoto、T.Iwasaka、H.Matsuda：“针对 Isk (MinK) 和过度表达的反义寡脱氧核苷酸对心脏延迟整流 K^ 电流的抑制

Y.Shigeyoshi, W.Fu, Y.Chen, K.Yagita, T.Takumi, P.Schotland, A.Sehgal, H.Okamura: "Restoration of circadian behavioral rhythms in a period null Drosophila mutant (perO) by mammalian Period homologues mPer1 and mPer2"Genes Cells. (in press).

Y.Shigeyoshi、W.Fu、Y.Chen、K.Yagita、T.Takumi、P.Schotland、A.Sehgal、H.Okamura：“通过哺乳动物周期恢复无效果蝇突变体 (perO) 的昼夜行为节律

T. Takumi: "A rapid cDNA cloning in the post-genome era."Methods Mol. Biol.. (in press).

T. Takumi：“后基因组时代的快速 cDNA 克隆。”Methods Mol。