Establishiment of calcium influx and detection system in the cilia

纤毛钙内流及检测系统的建立

• 批准号: 24659089
• 负责人: YOKOYAMA Takahiko
• 金额: $ 2.5万
• 依托单位: Kyoto Prefectural University of Medicine
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Challenging Exploratory Research
• 财政年份: 2012
• 资助国家: 日本
• 起止时间: 2012-04-01 至 2014-03-31
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-24659089/
• 关键词: 繊毛; カルシウム; シグナル

To establish a system that can get an influx of calcium and detect calcium in the cilia, we made channel-rhodopsin and cameleon(YC3.6) constructs linked to several cilia localization signals.These signals include Arl13B, Fiblocystin C-terminal and nphp3 N terminal. YC3.6 linked NPHP3 N-terminal and Arl13B with palmitoylation site successfully located to the cilia.Channel-rhodopsin linked to Arl13B also located to the cilia without palmitoylation site, suggesting that membrane protein such as channel-rhodopsin does not require palmitoylation to be localized to the cilia.We gave photo-stimulation to cells introduced channel-rodopsin. Unfortunately, calcium was not detected in these cells. To get effective calcium influx, an amplifier system may be required.

为了建立一个能够获得钙离子内流并检测纤毛中钙离子的系统，我们将通道-视紫红质和cameleon（YC 3.6）构建成连接有几个纤毛定位信号的结构，这些信号包括Ar 113 B，Fibrocystin C-末端和nphp 3 N-末端。YC 3.6成功地将NPHP 3的N端和带有棕榈酰化位点的Ar 113 B连接到纤毛上，而与Ar 113 B连接的通道视紫红质也定位在纤毛上，而不带有棕榈酰化位点，这表明膜蛋白如通道视紫红质不需要棕榈酰化就能定位到纤毛上。不幸的是，在这些细胞中没有检测到钙。为了获得有效的钙流入，可能需要放大器系统。

项目成果

繊毛病としての嚢胞性腎疾患

囊性肾病是一种纤毛病

• 发表时间: 2013
• 作者: 辻琢磨;横山尚彦;横山尚彦
• 通讯作者: 横山尚彦

A three step process of Nphp3 ciliary localization

Nphp3 纤毛定位的三步过程

• 发表时间: 2013
• 作者: Shiba D;Nakata K;Fukui H;Kobayashi D and Yokoyama T
• 通讯作者: Kobayashi D and Yokoyama T

A three-step process of Nphp3 ciliary localization.

Nphp3 纤毛定位的三步过程。

• 发表时间: 2013
• 作者: Shiba D;Nakata K;Fukui H;Kobayashi D and Yokoyama T.
• 通讯作者: Kobayashi D and Yokoyama T.

繊毛は体の「右と左」を決める

纤毛决定身体的“左右”侧

• 发表时间: 2013
• 期刊: 京都府立医科大学雑誌
• 作者: 小林大介;横山尚彦
• 通讯作者: 横山尚彦

横山研究室

横山实验室