| 研究課題/領域番号 |
24K23243
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| 研究種目 |
研究活動スタート支援
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| 配分区分 | 基金 |
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| 審査区分 |
0704:神経科学、ブレインサイエンスおよびその関連分野
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| 研究機関 | 京都大学 |
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研究代表者 |
劉 品吾 京都大学, 医学研究科, 特定研究員 (60886563)
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| 研究期間 (年度) |
2024-07-31 – 2026-03-31
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| 研究課題ステータス |
交付 (2024年度)
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| 配分額 *注記 |
2,860千円 (直接経費: 2,200千円、間接経費: 660千円)
2025年度: 1,430千円 (直接経費: 1,100千円、間接経費: 330千円)
2024年度: 1,430千円 (直接経費: 1,100千円、間接経費: 330千円)
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| キーワード | CaMKII / glutamate receptor / synaptic plasticity / LLPS / AMPAR / NMDAR / GluSnFR / super-resolution imaging |
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| 研究開始時の研究の概要 |
Synaptic plasticity, crucial for memory and other brain functions, involves synaptic proteins with unclear regulation. We propose that synaptic protein condensates formed via liquid-liquid phase separation (LLPS) are key to synaptic plasticity.
This study aims to explore memory mechanisms through LLPS in vivo. By using photoactivatable proteins, we will manipulate LLPS in living cultures and observe neuron network changes with light stimulation.
The findings could uncover mechanisms of memory formation and extinction, potentially leading to therapies for disorders like dementia and PTSD.
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| 研究実績の概要 |
In this project, we aim to understand how memory formation is regulated by the condensation of synaptic proteins through a process called liquid-liquid phase separation (LLPS). So far, we have focused on developing tools allowing us to control LLPS inside living neurons using light. We have successfully conducted a preliminary test of a fluorescence-based glutamate sensor in cultured neurons, confirming that it works at a basic level. However, we have encountered problems in creating light-controllable versions of a protein called CaMKII. Because the modified protein is very large, it has been difficult to produce it efficiently in common lab cell system used for protein testing. Despite these difficulties, our early results show that we are moving toward the goals set for this project.
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| 現在までの達成度 (区分) |
現在までの達成度 (区分)
3: やや遅れている
理由
At present, we are continuing the development of molecular tools for Aim 1 of the project. Preliminary experiments using the fluorescence-based glutamate sensor in neuronal cultures have confirmed that the sensor functions as intended, which is a positive step. However, the photoactivatable CaMKII constructs have shown poor production levels in the test cells. This suggests that the size and complexity of the engineered proteins may need to be optimized further before moving forward to light-controlled experiments in neurons.
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| 今後の研究の推進方策 |
Looking ahead, we will work on improving the design of our photoactivatable proteins. In addition to adjusting the current constructs, we plan to explore other methods for making proteins responsive to light, potentially by reducing some of the tags that were initially added as reporters. These improvements will hopefully make protein expression in cells easier and allow us to proceed with further experiments in neurons. In parallel, we will continue validating the use of iGluSnFR to monitor changes in synaptic communication under controlled conditions.
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