| Project/Area Number |
23K26919
|
|---|
| Project/Area Number (Other) |
23H02226 (2023)
|
|---|
| Research Category |
Grant-in-Aid for Scientific Research (B)
|
| Allocation Type | Multi-year Fund (2024)
Single-year Grants (2023) |
|---|
| Section | 一般 |
|---|
| Review Section |
Basic Section 39050:Insect science-related
|
|---|
| Research Institution | Juntendo University |
|---|
Principal Investigator |
グセフ オレグ 順天堂大学, 大学院医学研究科, 特任先任准教授 (30711999)
|
|---|
| Co-Investigator(Kenkyū-buntansha) |
黄川田 隆洋 国立研究開発法人農業・食品産業技術総合研究機構, 生物機能利用研究部門, グループ長 (60414900)
|
|---|
| Project Period (FY) |
2023-04-01 – 2027-03-31
|
| Project Status |
Granted (Fiscal Year 2024)
|
| Budget Amount *help |
¥18,590,000 (Direct Cost: ¥14,300,000、Indirect Cost: ¥4,290,000)
Fiscal Year 2026: ¥4,290,000 (Direct Cost: ¥3,300,000、Indirect Cost: ¥990,000)
Fiscal Year 2025: ¥4,290,000 (Direct Cost: ¥3,300,000、Indirect Cost: ¥990,000)
Fiscal Year 2024: ¥5,200,000 (Direct Cost: ¥4,000,000、Indirect Cost: ¥1,200,000)
Fiscal Year 2023: ¥4,810,000 (Direct Cost: ¥3,700,000、Indirect Cost: ¥1,110,000)
|
|---|
| Keywords | anhydrobiosis / snRNAseq / brain / insect / cell populations / brain cell populations / neurons / single cell RNAseq / neuronal cells / cytoprotective molecules |
|---|
| Outline of Research at the Start |
This study aims to elucidate molecular and genetic mechanisms protecting against complete desiccation variety of cells in the brain of the sleeping chironomid and to identify compounds with neuroprotective effects for biomedical applications.
|
| Outline of Annual Research Achievements |
This study aims to elucidate molecular and genetic mechanisms protecting against complete desiccation cells in the brain of the sleeping chironomid and to identify compounds with neuroprotective effects for biomedical applications.
To reveal the complexity of cell types and their destiny during anhydrobiosis, we conducted detailed single-nuclei transcriptomics profiling of the chironomid brains in six time points via the cycle of desiccation-rehydration. During this year we have optimized the larvae dissection for brain extraction and adaptation of cell dissociation protocol. We made an initial reconstruction of cell population composition dynamics and adaptation using single-cell mRNA expression events in the brain of the chironomid through the cycle of desiccation and rehydration.
|
| Current Status of Research Progress |
Current Status of Research Progress
1: Research has progressed more than it was originally planned.
Reason
Based on the research plan we have successfully adapted the protocol of dissection of chironomid brain, followed by single-nuclei RNAseq analysis of six timed points of anhydrobiosis. As a result, we successfully profiled a total of 86 000+ nuclei. Due to rapid progress with T-2-T genome analysis of P. vanderplanki we achieved a total number of more than 600 genes per cell profiling. We identified key cell types by comparing P.v. and Drosophila data. Already on the current step of the study, we identified shifts in cell populations in response to desiccation and made the hypothesis about the specific vulnerability of some organelles to desiccation. We finally could indirectly confirm our predictions using analysis of Pv11 cell morphology. These results were published as a research paper.
|
| Strategy for Future Research Activity |
As planned, we will the next major step: analysis of metabolomic events associated with anhydrobiosis in the brain of the larvae, using both metaboles profiling and in silico combination of transcriptomic and proteomic data from the brain of P. vanderplaki. We expect to obtain a profile of changes of up to 400 metabolites in the brains of the larvae. We will further list prospective candidates for metabolites, proteins, and peptides with predicted neuroprotective effects on human neurons by combining our gene expression and metabolomic data, literature survey, and molecular effect simulation. At the end of the year, we expect to obtain a list of promising candidates for further in vitro (and in vivo in the future) experiments for validation of protective potential for mammalian neurons.
|
