2021 Fiscal Year Annual Research Report
Super-resolution imaging for synapse reveals structure-function correlation between sub-synaptic protein localization and synaptic plasticity
| Project/Area Number |
20K22685
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| Research Institution | Kyoto University |
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Principal Investigator |
劉 品吾 京都大学, 医学研究科, 研究員 (60886563)
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| Project Period (FY) |
2020-09-11 – 2022-03-31
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| Keywords | Super-resolution imaging / STORM / synaptic plasticity / memory formation |
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| Outline of Annual Research Achievements |
The aim of this study is to investigate the molecular mechanism underlying the memory formation. It has been reported that the synaptic proteins form sub-synaptic clusters upon the excitatory stimulation. Recently, we have reported the synaptic activity-dependent organization of nano-scale localization of postsynaptic proteins via liquid-liquid phase separation, which is critical for the regulation of synaptic plasticity. Especially we found that the representative glutamate receptors, AMPAR and NMDAR, had segregated each other in vitro, which would explain the potentiation of synapse during memory formation. However, observation for detailed structure of synaptic clusters requires dual-channel super-resolution microscopy. Thus, to expand our research, we set up the dual-color super-resolution microscopy with Nikon STORM system.
In the last year, we have established the single-color observation of this super-resolution system. In this year, we improved the efficacy of fluorophore blinking which is related with the resolution of final image, of the second color channel by optimizing the imaging buffer. Furthermore, we upgraded the laser power and modified the pass way of the laser beam. After those efforts, we successfully completed the setup for dual-color STORM imaging.
Using these improved systems, we will observe not only the combination of AMPAR and NMDAR, but also the other synaptic proteins with receptors, which reveals excitatory stimulation-triggered nano-scale reorganization of synaptic proteins to enhance synaptic transmission after learning.
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