2018 Fiscal Year Annual Research Report
Cell type-specific mechanisms of desiccation tolerance in the anhydrobiotic insect
| Project/Area Number |
18H02217
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| Research Institution | Institute of Physical and Chemical Research |
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Principal Investigator |
グセフ オレグ 国立研究開発法人理化学研究所, 科技ハブ産連本部, ユニットリーダー (30711999)
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| Project Period (FY) |
2018-04-01 – 2021-03-31
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| Keywords | scRNAseq / Pv11 cells / the sleeping chironomid / 10xGenomics platform / LEA proteins |
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| Outline of Annual Research Achievements |
Using 10XGenomics platform we will analyze 8000-1000 single cells transcriptomic profile of Pv11 cell line on the different time points: control, trehalose treatment for 24h (pre-conditioning) and trehalose treatment for 48h (ready to complete desiccation) Using new version on the Pv genome (MidgeBase 2.0 with N50 is 1.8M bp, four chromosome-level contigs in total) ad the new improved annotation we identified variety of Pv11 cells sub-populations and characterize its response to desiccation. We found that Pv11 population contains of six major subtypes of cells that have clearly different response to trehalose treatment. We also identified specific sets of protective genes (including Lea proteins, hsp, etc.) specific for each sub-type. Specifically, we assume that initial cell life cycle status is one of the key definition of success of anhydrobiosis. Also, we confirmed that protective genes contained in ARId regions of genomes and responsive to desiccation is actually how cell type-specific pattern in expression. The strongest cell-type specific patterns were observed for LEA genes and PIMT genes.
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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
We have successfully adapted Pv11 cells for single cell sequencing. Initial challenge was to maintain viable cells under trehalose treatment and it was sussesfully achieved. We also confirmed our initial hypothesis about heterogeneity of Pv11 cells and its possible strong influence of the capacity of Pv11 line to anhydrobiosis. We have also observed significant specialization of protective anhydrobiosis-related genes expression depending on sub-type of Pv11 cells. In total our observation provided additional evidence of strong cell type-specific nature of anhydrobiosis mechanism.
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| Strategy for Future Research Activity |
According to initial plan, Using 10xGenomics Chromium single cell sequencing platform, we will evaluate
gene expression in up to 20 000 cells, derived from different organs and tissues the larvae (to be provided by Dr. Kikawada) on its different time points of desiccation and rehydration (i.e. cycle of
anhydrobiosis). Updating MidgeBase 2.0 database with single cell data. Result: we will identify transcriptional profiles of major primary Pv cell types and cell type-specific groups of ARId-located and other protective genes associated with anhydrobiosis
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