| Project/Area Number |
22K20702
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| Research Category |
Grant-in-Aid for Research Activity Start-up
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| Allocation Type | Multi-year Fund |
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| Review Section |
0704:Neuroscience, brain sciences, and related fields
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| Research Institution | Institute of Physical and Chemical Research |
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Principal Investigator |
Phasuk Sarayut 国立研究開発法人理化学研究所, 生命機能科学研究センター, 特別研究員 (70961580)
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| Project Period (FY) |
2022-08-31 – 2024-03-31
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| Project Status |
Completed (Fiscal Year 2023)
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| Budget Amount *help |
¥2,860,000 (Direct Cost: ¥2,200,000、Indirect Cost: ¥660,000)
Fiscal Year 2023: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000)
Fiscal Year 2022: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000)
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| Keywords | m6A / YTHDF3 / YTHDF1 / Place cells / Calcium imaging / Two photon microscopy / Spatial memory / Virtual reality / Place Cells / Calcium Imaging / Two Photon Microscopy / Spatial Memory / Virtual Reality |
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| Outline of Research at the Start |
We will genetically control YTHDF3 levels in hippocampal CA1 excitatory neurons. We will then observe hippocampal neuronal activity in mice performing a learning task by 2p calcium imaging, and examine how m6A affects the activity of place cells. Elucidation of the role of protein spatiotemporal translation mediated by m6A in spatial learning is expected to greatly contribute to the development of new treatments for dementia and other diseases.
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| Outline of Final Research Achievements |
In this study, we used a conditional knockout (cKO) mouse model lacking the m6A reader protein YTHDF3 in excitatory neurons. An adeno-associated virus (AAV) with GCaMP6f, a green calcium fluorescence sensor, was introduced into the hippocampal CA1 to monitor neuronal activity. Two weeks post-injection, 3-month-old mice underwent optical window implantation. After a one-month recovery, the mice were placed on a Styrofoam treadmill under a two-photon microscope with a secured head plate. They performed virtual reality tasks on a linear track with water rewards.
We monitored over 1,000 neurons in the hippocampal CA1 using in vivo two-photon (2p) calcium imaging. Running speed and licking behavior were recorded and matched with visual cues. We identified cellular mechanisms of spatial learning-induced place cell formation without m6A function, observing dendritic spine activity. This study demonstrated the benefits of combining VR tasks with 2p calcium imaging, achieving our research goals.
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| Academic Significance and Societal Importance of the Research Achievements |
Our research with a cKO mouse model lacking YTHDF3 revealed key insights into spatial learning and memory. It highlighted m6A methylation's role in synaptic plasticity and suggested therapeutic targets. Integrating VR technology enhances experimental accuracy and innovation.
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