2022 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 | Aging / ATP / LPS / Neurodegeneration |
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| Outline of Annual Research Achievements |
The project is going according to plan without significant problems. The data obtain so far are as followed:
1) From the original 4 human iPSC lines, 3 have been successfully cultured, expanded, and cryopreserved.
2) The cell proliferation rate and viability of iPSC from 20 y/o, 40 y/o and 80 y/o individuals have been analyzed and showed significant reduction in the number of colonies, number of cells and cell viability in older iPSCs compared to younger iPSCs.
3) The intracellular levels of ATP in the various iPSC lines have been measured by in vitro bioluminescence assay and revealed a substantial reduction in ATP levels from 80 y/o iPSCs compared to 20 or 40 y/o iPSCs.
4) Reduction in intracellular levels of ATP has also been confirmed in mouse DRG primary cultures from old animals (52 weeks) compared to younger animals (8 weeks), supporting our observations in human iPSCs.
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| Current Status of Research Progress |
Current Status of Research Progress
2: Research has progressed on the whole more than it was originally planned.
Reason
The progress of the project is going according to plan and the initial characterization of the effect on aging using iPSCs has been achieve by measuring cell profiliferation, viability and ATP levels.
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| Strategy for Future Research Activity |
I am currently analyzing liquid phase separation by FRAP in both human iPSC lines and mouse DRG neurons, as well as quantifying the number of active mitochondria in old samples versus young samples by confocal imaging using TMRE.
Additionally, I am extracting genomic DNA and mitochondrial DNA from iPSCs and DRGs to analyze their DNA methylation levels and determine a potential correlation between genetic clock, mitochondria activity and ATP production, and protein condensation.
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| Causes of Carryover |
The carry over was done because of delay in obtaining cell lines and some reagents.
The carry over budget will be used to performed DNA methylation assay, ATP measurement and live imaging to complete the remaining experiments. Also to attend meeting for presentation and publication fees.
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