2018 Fiscal Year Research-status Report
Functional analysis of an RNA methyltransferase in regulation of neural stem cell fate
| Project/Area Number |
18K14722
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| Research Institution | Institute of Physical and Chemical Research |
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Principal Investigator |
呉 泉 国立研究開発法人理化学研究所, 生命機能科学研究センター, 基礎科学特別研究員 (60812766)
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| Project Period (FY) |
2018-04-01 – 2021-03-31
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| Keywords | Brain development / RNA methltransferase / Single cell analysis |
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| Outline of Annual Research Achievements |
The purpose of this study is to clarify the function of Fibrillarin (Fbl), a RNA methyltransferase, in brain development. I have confirmed that knockout of Fbl caused microcephaly previously. In this year, to determine whether p53-dependent apoptosis is the cause of microcephaly of mutant brains, I crossed Fbl cKO mice with p53-/- and obtained Fbl flox/flox Emx1-Cre p53-/- double knockout mice (dKO). I found that brains of dKO were also smaller than control mice, indicating that microcephaly could not only explained by the apoptosis of neural stem cells (NSCs). To further examine the impact of Fbl on NSCs, I investigated cortical organization of dKO at E17. Notably, the number of neurons were significantly reduced when compared with control brains. To investigate to what extent Fbl affects temporal identity, I performed transcriptome analysis from isolated single cells with different genotypes along developmental time. Bioinformatics analysis of these data indicated that Fbl is required for temporal fate transition of NSCs. To identify the targets of Fbl, I also performed ribosome profiling using dKO mice brains and have already identified several candidates that might be controlled by Fbl.
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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
In this year, we analyzed Fbl mutant mice by using different approaches at both mRNA (single cell analysis) and protein levels (ribosome profiling). We proved that Fbl is an important regulator for the temporal fate transition of NSCs. Importantly, I have already identified potential targets of Fbl.
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| Strategy for Future Research Activity |
In the next year, I will functional analysis identified targets of Fbl by using siRNA or genetic mutant mice. In the same time, I will find out how Fbl affects the translation of these genes specifically.
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| Causes of Carryover |
The mutant mice which were used for the sequencing expeirment was not as much as expected, therefore, the money for sequencing was remained.
The remained money will be used in next fiscal year for sequencing.
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