| Budget Amount *help |
¥4,810,000 (Direct Cost: ¥3,700,000、Indirect Cost: ¥1,110,000)
Fiscal Year 2017: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000)
Fiscal Year 2016: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000)
Fiscal Year 2015: ¥1,950,000 (Direct Cost: ¥1,500,000、Indirect Cost: ¥450,000)
|
|---|
| Outline of Final Research Achievements |
For making aquaporin (AQP) 4 knock down, we used Lentiviral RNAi system of Invitrogen corporation. Finally, we obtained AQP 4 knockdown model with pLenti4/BLOCK-IT/Expression Construct and plasmid of pLP1、pLP2、pLP/VSVG、and pENTER-gus. We confirmed that AQP4 knockdown that was stronger than our earlier experiment. We constructed hAQP4-M23 and hAQP4-M1 vector and transfected into C6 cells. We confirmed the overexpression of hAQP4-M1 and hAQP4-M23 in C6 cells. Moreover, we constructed permanent cell line of hAQP4-M23-overexpressed-cells of C6.In hypoxic condition, we tried to decide apoptosis or necrosis with flow cytometry. For optimal oxygen concentration, we experimented cell culture of astrocyte and SH-SY5Y in hyperoxia. We confirmed cell damage of hyperoxia in this condition. We try to continue neuron protection of various condition and drugs using RNAi technique.
|
