2017 Fiscal Year Research-status Report
The enzymatic activity of Cathepsin E in Abeta degradation
| Project/Area Number |
17K17093
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| Research Institution | Kyushu University |
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Principal Investigator |
倪 軍軍 九州大学, 歯学研究院, 助教 (00783953)
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| Project Period (FY) |
2017-04-01 – 2019-03-31
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| Keywords | Alzheimer's disease / Microglia / Cathepsin E |
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| Outline of Annual Research Achievements |
1, The CatE overexpressed microgla has been established based on the pEGFP-N3 plasmid. The overexpression efficiency has been checked by both RT-PCR and Western blotting.
2, The clearance activity of CatE has been checked by using CatE overexpressed microglia. The phagocytotic activity of microglia were not changed between WT and CatE overexpressed microglia, however, the digestion efficiency of uptaked Abeta were significantly increased in CatE overexpressed microglia.
3, The APP NL-G-F/NL-G-F/CatE-/- mice were picked up and maintained for future experiments.
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| Current Status of Research Progress |
Current Status of Research Progress
3: Progress in research has been slightly delayed.
Reason
1, The CatE knock-out microglia were not obtained using Crispr-cas9 system using CatE target sequence inserted pCas-Guide, because of the low efficiency of transfection.
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| Strategy for Future Research Activity |
1, Using the APP NL-G-F/CatE-/- mice brain samples to checked the Abeta accumulation in mice cortex and hippocampus.
2, Memory tests of APP NL-G-F/CatE-/- mice using Y-maze test, novel object recognition test and step-through passive avoidance test, when the mice number is enough.
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| Causes of Carryover |
The incurring amount will be used for mouse behavior test and in-vivo experiments.
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