|Budget Amount *help
¥17,030,000 (Direct Cost: ¥13,100,000、Indirect Cost: ¥3,930,000)
Fiscal Year 2021: ¥4,680,000 (Direct Cost: ¥3,600,000、Indirect Cost: ¥1,080,000)
Fiscal Year 2020: ¥4,160,000 (Direct Cost: ¥3,200,000、Indirect Cost: ¥960,000)
Fiscal Year 2019: ¥4,160,000 (Direct Cost: ¥3,200,000、Indirect Cost: ¥960,000)
Fiscal Year 2018: ¥4,030,000 (Direct Cost: ¥3,100,000、Indirect Cost: ¥930,000)
|Outline of Final Research Achievements
We established a correlative live-cell imaging system by combining high-speed atomic force microscopy (HS-AFM) and confocal laser-scanning microscopy and revealed the detailed morphological changes in the plasma membrane and protein localizations during the clathrin-mediated endocytosis. CIP4, which has membrane-deforming activity, is recruited to the plasma membrane via small G protein Cdc42, and promotes the assembly of actin, which induces a small membrane bulge near the clathrin pit. We also made an ultra-thin stretching cell chamber using polydimethylsiloxane and successfully installed into the above-mentioned correlative imaging system together with a thin stretching device. By using this live-cell stretching system, we analyzed how one-dimensional stretching stimulus affected the progress of clathrin-mediated endocytosis, and identified the tension-dependent steps in the initiation and closing steps.