2021 Fiscal Year Research-status Report
ATP-dependent liquid phase separation during aging and neurodegeneration
| Project/Area Number |
21K06400
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| Research Institution | Okinawa Institute of Science and Technology Graduate University |
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Principal Investigator |
Guillaud Laurent 沖縄科学技術大学院大学, 分子神経科学ユニット, グループリーダー (90596222)
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| Project Period (FY) |
2021-04-01 – 2024-03-31
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| Keywords | Liquid phase separation / Aging / Mitochondria / ATP / Neurodegeneration |
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| Outline of Annual Research Achievements |
We performed preliminary experiments on mouse primary neurons and human motor neurons from healthy individual and ALS patients currently available in lab. We measured mitochondria activity, ATP concentration and liquid phase separation of cGFP in mouse and human neurons and showed a significant reduction in ATP concentration in ALS neurons compared to healthy neuron, correlated to a decrease in LPS. We also tested the effect of NMN chronic treatment on mouse neurons and showed that 48h treatment induced increase of ATP in treated neurons compared to untreated neurons.
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| Current Status of Research Progress |
Current Status of Research Progress
3: Progress in research has been slightly delayed.
Reason
We had delay in selecting and acquiring human iPSC lines during the first year of the project but all the necessary lines are going to be purchase in the coming fiscal year.
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| Strategy for Future Research Activity |
After acquisition and delivery of iPSC lines of different ages we will initiate their expansion and differentiation into motor neurons. After successful differentiation we will monitor and quantify mitochondria number, size and activity in motor neurons obtained from iPSCs of different ages. Concomitantly we will measure the axo-terminal and cell body cytosolic fluid phase by FRAP in motor neurons transduced with AAV-cGFP and expressing cytosolic GFP. And measure intracellular ATP concentration in neurons expressing PercevalHR (ATP fluorescent sensor) and by in vitro bioluminescence assay.
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| Causes of Carryover |
We had delay to select and acquire the necessary human iPSC lines during the first year and the different lines will be purchased this coming fiscal year.
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