Study of IP3 receptor/Ca^<2+> signaling in neural plasticity and brain development and differentiation

IP3受体/Ca^2信号在神经可塑性和脑发育分化中的研究

• 批准号: 15100006
• 负责人: MIKOSHIBA Katsuhiko
• 金额: $ 77.04万
• 依托单位: The Institute of Physical and Chemical Research (2007)The University of Tokyo (2003-2006)
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (S)
• 财政年份: 2003
• 资助国家: 日本
• 起止时间: 2003 至 2007
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-15100006/
• 关键词: IP_3 receptor; Ca^<2+> signaling; neuronal plasticity; endoplasmic reticulum; redox sensor; Ip_ indicator; Na+K+-ATPase; 2-APB; 2型,3型IP_3レセプターダブルノックアウトマウ; IP_3レセプター; プルキンエ細胞; 酸・塩基平衡; ナトリウム重炭酸イオン共輸送体1; カルシウム振動; カルシウム; カルシウムチャンネル; 小胞体; ERp4

IP_3 is a second messenger to release Ca^<2+> from internal stores. We discovered that P400 which is deficient in Purkinje-neuron-degenerating mutant is IP_3 receptor (IP_3R). IP_3R /P400 is found to be localized on endoplasmic reticulum (ER). Purified IP_3 receptor (IP_3R) works as Ca^<2+> release channel and works as Ca^<2+> oscillator. IP_3R function is regulated by phosphorylation (by PKC, CaMK II, PKG) suggesting that IP_3R works as a cross-talk station between Ca^<2+> signaling and phosphorylation. IP_3R has unique properties: 1) IP_3R is functional even though it is fragmented by proteases into several pieces; 2) IP_3R allosterically and dynamically changes its form reversibly; _3) CryoEM study shows that IP_3R contains multiple cavities; 4) ER forms a meshwork and vesicular ER containing IP_3R moves rapidly along microtubles using kinesin motor. We succeeded in crystallization of IP_3 binding core and suppressor sequence. They show unique structure and interact each other to re … More cognize IP_3. Studies on the role of IP_3R during development show that IP_3R is involved in fertilization and is essential for determination of dorsoventral axis formation. IP_3R is involved in neuronal plasticity. Double homozygous mutant of IP_3R2 and IP_3R_3 shows deficit of exocrine secretion. ERp44 works as a redox sensor in the ER and regulates IP_3R1 activity. We developed IP_3 indicator (named as IRIS) using the IP_3 binding core. We discovered that IP_3 not only releases Ca^<2+>, but also releases IRBIT. IRBIT binds to the same site as IP_3. IRBIT works only when it is phosphorylated. It regulates frequency and amplitude of Ca^<2+> oscillation generated by IP_3R. In addition, phosphorylated IRBIT binds to pancreas type Na, Bicarbonate co-transporter 1 (NBC1) and works as a third messenger to enhance NBC1 which regulates pH inside cells. Therefore, signaling pathway may be modified to be as follows: [ extracellular signal →IP_3→IP_3R→Ca^<2+> release, and IRBIT release→NBC1 activation, and Ca^<2+> oscillation modification ]. Less

IP_3是从内部商店释放CA^<2+>的第二个使者。我们发现在Purkinje-Neuron脱生突变体中具有确定性的P400是IP_3受体（IP_3R）。发现IP_3R /P400位于内质网（ER）上。纯化的IP_3受体（IP_3R）用作CA^<2+>释放通道，并用作CA^<2+>振荡器。 IP_3R函数受磷酸化（通过PKC，CAMK II，PKG）调节，这表明IP_3R是CA^<2+>信号传导和磷酸化之间的串扰站。 IP_3R具有唯一的属性：1）IP_3R具有功能性，即使它被蛋白酶分成多个部分； 2）IP_3R在变构和动态上可逆地更改其形式； _3）冷冻研究表明IP_3R包含多个咖啡馆； 4）ER使用驱动蛋白电动机形成含有IP_3R的网状功能和囊泡ER沿微公开迅速移动。我们成功地结晶了IP_3结合核心和抑制序列。它们显示独特的结构并相互交互以进行……更多的IP_3。 IP_3R在开发过程中的作用的研究表明，IP_3R参与了受精，对于确定背腹轴的形成至关重要。 IP_3R参与神经元可塑性。 IP_3R2和IP_3R_3的双纯合突变体显示外分秘密的不足。 ERP44在ER中充当氧化还原传感器，并调节IP_3R1活动。我们使用IP_3绑定核心开发了IP_3指示器（称为IRIS）。我们发现IP_3不仅发行了Ca^<2+>，而且还发布了Irbit。 IRBIT与IP_3的站点结合。仅在将其磷酸化时才能起作用。它调节IP_3R生成的Ca^<2+>振荡的频率和放大器。此外，磷酸化的含IRBIT与Na型胰腺，碳酸氢盐共转运蛋白1（NBC1）结合，并用作第三个使者，以增强调节细胞内pH的NBC1。因此，信号通路可以修改为如下：[细胞外信号→IP_3→IP_3R→Ca^<2+>释放，而IRBIT释放→NBC1激活以及Ca^<2+>振荡修改]。较少的

项目成果

Organelles containing inositol trisphosphate receptor type 2 in adrenal medullary cells

• DOI: 10.2170/physiolsci.rp006406
• 发表时间: 2006-12-01
• 期刊: JOURNAL OF PHYSIOLOGICAL SCIENCES
• 影响因子: 2.3
• 作者: Endo，Yutaka;Harada，Keita;Inoue，Masumi
• 通讯作者: Inoue，Masumi

Visuatization of inositol 1, 4, 5-trisphosphate receptor by atomic force microscopy.

通过原子力显微镜观察肌醇 1,4,5-三磷酸受体。

• 发表时间: 2006
• 期刊: Neuroscience Letters 391
• 作者: Suhara;W.;Kobayashi;M.;Sagara;H.;Hamada.;K.;Goto;T.;Fujimoto;I.;Torimitsu;K.;Mikoshiba;K.
• 通讯作者: K.

Visualization of inositol 1,4,5-trisphosphate receptor by atomic force microscopy

通过原子力显微镜观察肌醇 1,4,5-三磷酸受体

• 发表时间: 2006
• 期刊: Neuroscience Letters 391
• 作者: Suhara;W.
• 通讯作者: W.

Human malarial parasite,Plasmodium falciparum,displays capacitative calcium entry:2-aminoethyl diphenylborinateblocks the signal transfuction pathway of melatonin action on the P.falciparum cell cycle.

人类疟原虫恶性疟原虫表现出电容性钙内流：2-氨基乙基二苯硼酸酯阻断褪黑素对恶性疟原虫细胞周期作用的信号转导途径。

• 发表时间: 2007
• 期刊: Journal Pineal Research. 43
• 作者: Beraldo;H.F.;et. al.
• 通讯作者: et. al.

Optimization of Transfection Conditions for Gene Expression, siRNA Knock-Down, and Live Cell Imaging Using FuGENE(R) HD Transfection Reagent.

使用 FuGENE(R) HD 转染试剂优化基因表达、siRNA 敲低和活细胞成像的转染条件。

• 发表时间: 2008
• 期刊: BIOCHEICA 1
• 作者: Kawaai K;Ishida S;Matsu-ura T;Kuroda Y;Ogawa N;Tashiro T;Enomoto M;Hisatsune C;Mikoshiba;K.
• 通讯作者: K.