| Budget Amount *help |
¥4,160,000 (Direct Cost: ¥3,200,000、Indirect Cost: ¥960,000)
Fiscal Year 2015: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000)
Fiscal Year 2014: ¥1,170,000 (Direct Cost: ¥900,000、Indirect Cost: ¥270,000)
Fiscal Year 2013: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000)
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| Outline of Final Research Achievements |
The neurotoxicity of anesthetics on the developing brain has drawn the attention of anesthesiologists. Previous studies have shown that prolonged exposure to anesthetics causes apoptosis. Exactly when the apoptotic cascade starts in the brain remains uncertain. We describe the development of a continuous monitoring system to detect caspase-3 activation using an in vivo model. Brain slices from neonatal SCAT3 transgenic mice with a heterozygous genotype (n = 20) were used for the monitoring of caspase-3 cleavage. SCAT3 is a fusion protein of ECFP and Venus connected by a caspase-3 cleavable peptide, DEVD. A specimen from the hippocampal CA1 sector was mounted on a confocal laser microscope and was continuously superfused with propofol. We observed a shift in the histogram toward the right over time, indicating caspase-3 activation at five hours in the propofol. Thus, real-time FRET imaging was capable of identifying the onset of apoptosis triggered by propofol in neonatal brain slices.
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