SDS-digested freeze-fracture replica labeling(SDS-FRL) was innovated as a suitable technique to reveal the fine structure of plasma membrane and quantitative localization of biomolecules over the membrane structure. However, it was difficult to apply this

创新了 SDS 消化冷冻断裂复制标记（SDS-FRL）作为揭示质膜精细结构和生物分子在膜结构上定量定位的合适技术。

• 批准号: 21500311
• 负责人: FUKAZAWA Yugo
• 金额: $ 2.91万
• 依托单位: Nagoya University (2011)National Institute for Physiological Sciences (2009-2010)
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2009
• 资助国家: 日本
• 起止时间: 2009 至 2011
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-21500311/
• 关键词: 凍結割断レプリカ; 電子顕微鏡; 分子局在; 細胞膜

SDS-digested freeze-fracture replica labeling(SDS-FRL) was innovated as a suitable technique to reveal the fine structure of plasma membrane and quantitative localization of biomolecules over the membrane structure. However, it was difficult to apply this technique to molecules immobilized on protoplasmic faces(P-face) quantitatively. Thus, in this research project, I tried to optimize labeling protocols of SDS-FRL. The optimization was carried out at steps of SDS treatment, pretreatment and blocking of replicated membranes as well as components on each labeling solutions. As a result, although details of labeling protocol differ among experiments, new protocol provided high-sensitive labeling not only for molecules on exoplasmic faces but also for those on P-faces. Localization data obtained by these investigations were published as original papers.

SDS-FRL是一种创新的技术，可用于揭示质膜的精细结构和在膜结构上定量定位生物分子。然而，该技术难以定量地应用于固定在原生质面（p面）上的分子。因此，在本研究项目中，我尝试优化SDS-FRL的标记方案。在SDS处理、预处理和阻断复制膜以及各标记溶液上的组分等步骤进行优化。因此，尽管不同实验的标记方案细节不同，但新方案不仅对外质面分子提供了高灵敏度的标记，而且对p面分子也提供了高灵敏度的标记。这些调查获得的定位数据作为原始论文发表。

项目成果

The enhancement of PSD size and AMPA receptor density in the central amygdala in rat modle of neuropathic pain

神经病理性疼痛模型大鼠杏仁核中央PSD大小和AMPA受体密度的增强

• 发表时间: 2009
• 作者: Dong D;Fukazawa Y;Shigemoto R
• 通讯作者: Shigemoto R

Electron microscopy of biological membrane. Encyclopedia of Biophysics

生物膜的电子显微镜。

• 发表时间: 2012
• 作者: Fujimoto T;Fukazawa Y
• 通讯作者: Fukazawa Y

定量的分子局在解析と計算論的手法によるシナプス構造-機能連関の解析

使用定量分子定位分析和计算方法分析突触结构-功能关系

• 发表时间: 2011

Presynaptic, postsynaptic, and morphological determinants of signal transmission at the retinogeniculate synapse

视网膜突触信号传递的突触前、突触后和形态决定因素

• 发表时间: 2010
• 作者: Matsui K;Budisantoso T;Kamasawa N;Fukazawa Y;Shigemoto R
• 通讯作者: Shigemoto R

In vivo remodeling of postsynaptic glutamate receptor or ganization revealed by freeze-fracture replica labeling

通过冷冻断裂复制标记揭示突触后谷氨酸受体或组织的体内重塑

• 发表时间: 2009
• 作者: Fukazawa Y;Itakura M;Takahashi T;SaitohY;Inokuchi K;Molnar E;Shigemoto R
• 通讯作者: Shigemoto R