2020 Fiscal Year Research-status Report
ATP-dependent coupling of pre- and post-synaptic organelles movements at central synapses and its implication in synaptic plasticity and transmission
| Project/Area Number |
18K06494
|
|---|
| Research Institution | Okinawa Institute of Science and Technology Graduate University |
|---|
Principal Investigator |
Guillaud Laurent 沖縄科学技術大学院大学, 細胞分子シナプス機能ユニット, 研究員 (90596222)
|
|---|
| Project Period (FY) |
2018-04-01 – 2022-03-31
|
| Keywords | Liquid phase separation / Synapse / Mitochondria / ATP / Neurodegeneration |
|---|
| Outline of Annual Research Achievements |
For the third year my research project has progressed according to plan but with a slight delay due to my move to a new research unit. However this move allowed me to also expand my work to motor neurons and related neurodegenerative pathologies.
1) I have now completed the in vitro analysis of 6 proteins involved in Parkinson and Alzheimer diseases and quantified the number and size of their condensates at 37C and in the presence or absence of ATP. I also determined the threshold concentration of ATP required to prevent liquid phase separation of each of these proteins. 2) I have also performed liquid phase separation in vitro assay on 2 additional proteins implicated in ALS disease, and demonstrated that they also undergo ATP-dependent condensation/decondensation at 37C. 3) I have also performed preliminary experiments on cultured motor neurons (derived from human iPSC and/or mouse ES cells) and show that cytosolic GFP liquid phase separation follow the same mitochondria/ATP-dependent mechanism as I previously reported in giant glutamatergic terminals, suggesting a common and conserve LPS regulation mechanism among different neuronal populations. 4) Additionally I also analyzed LPS in cultured human glutamatergic neurons (derived from iPSCs) using FRAP, as well as quantified and compared ATP concentration in normal and Alzheimer cultured neurons by in vitro luminescence assay. 5) I have written and prepared the initial draft of the manuscript
|
| Current Status of Research Progress |
Current Status of Research Progress
3: Progress in research has been slightly delayed.
Reason
As I moved to a different unit at the end of 2019, my work has been slightly delayed. In addition I have also started to collect new data on different type of neurons (derived from human iPSCs or mouse ESCs) as well as on several proteins involved in ALS disease.
|
| Strategy for Future Research Activity |
My next research plan is to increase the number of data on motor neurons cytosolic phase separation and in vitro data on additional proteins involved in ALS or SMA.
Lastly I am planning to submit the manuscript in the first half of 2021.
|
| Causes of Carryover |
Due to my move to a new research unit and the delay in the preparation of the manuscript and the collection of additional and new set of data, I have requested carry over of my remaining budget to 2021.
the remaining budget will be used to cover the cost of reagents need for the revision of the manuscript and for the publication fee.
|
