研究実績の概要 |
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.
|
今後の研究の推進方策 |
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
|