| Budget Amount *help |
¥4,290,000 (Direct Cost: ¥3,300,000、Indirect Cost: ¥990,000)
Fiscal Year 2010: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000)
Fiscal Year 2009: ¥2,730,000 (Direct Cost: ¥2,100,000、Indirect Cost: ¥630,000)
|
|---|
| Research Abstract |
We were able to culture primary motor neurons from rat or mouse using dual chamber system. It enabled us to culture the synapse side and cell body side in the different medium. To improve the culture condition, we improved the condition of the medium. The axonal elongation has improved by adding the extract of skeletal muscles from mouse. We cultured motor neurons derived from human poliovirus (PV) receptor (hPVR) - transgenic (Tg) mouse on the dual chamber and added PV defective interfering particles coding GFP from the synapse side or the cell body side. When the virus was added from the cell body side, expression of GFP was observed at six hours after the addition. On the other hand, when the virus was added from the synapse side, expression of GFP had not been observed until twenty-four hours after the addition, while the expression was observed at forty-eight hours after the addition. These results suggest that the infection from the cell body side progresses differently to the infection from the synapse side. Next, we tried to optimize the four-dimensional imaging system for live fluorescent-cells. For long term culture under the microscope, we changed the medium into CO2 independent medium in a closed system. This enabled cells to live longer under the microscope. We tried various conditions of laser intensity, pinhole size, and number of Z slices for long term observation under the confocal laser microscope. However, we could not optimize the conditions for the long term observation as long as the confocal laser microscope was used.
|
