研究目的
Investigating the mechanisms of membrane remodeling during cleavage furrow formation and intercellular bridge elongation in cytokinesis using optogenetic control and super-resolution microscopy.
研究成果
The study demonstrates that cleavage furrow formation and intercellular bridge elongation involve spatially localized and coordinated endocytosis and exocytosis. RhoA activation induces actomyosin contractility and plasma membrane flow to the furrow, leading to localized endocytosis. Vesicles are trafficked to the bridge and undergo exocytosis, contributing to bridge elongation. Additionally, exocytosis at the front of daughter cells replenishes membrane lipids, supporting cell shape maintenance and movement.
研究不足
The optogenetic tool induces furrow formation and a structure resembling intercellular bridges but does not lead to abscission, possibly due to the bypass of steps prior to RhoA activation in native cytokinesis. The study is limited to macrophage cells, and the findings may not be directly applicable to other cell types.
1：Experimental Design and Method Selection:
Used a light-inducible dimerization (iLID) system to optically recruit LARG, a guanine nucleotide exchange factor (GEF) for RhoA, to the plasma membrane of RAW
2：7 macrophage cells, activate RhoA locally, and induce furrow formation. Sample Selection and Data Sources:
2 RAW
3：7 cells transfected with various constructs including iLID-CaaX, LARG-mTurq-SspB, and membrane markers like Venus-KRasCT and mCh-GL-GPI. List of Experimental Equipment and Materials:
2 Andor Revolution spinning disk confocal system, Leica DMI6000B microscope, Yokogawa CSU-X1 spinning disk, Andor iXon camera, Andor FRAPPA unit, Andor Dragonfly spinning disk confocal system for TIRF and SRRF-stream imaging.
4：Experimental Procedures and Operational Workflow:
Cells were photoactivated with 445 nm light to induce RhoA activation, imaged every 5 seconds, and analyzed for furrow formation and membrane dynamics.
5：Data Analysis Methods:
Fluorescence recovery after photobleaching (FRAP), intensity profile analysis, and custom algorithms for quantifying exocytosis events and membrane incorporation.
独家科研数据包，助您复现前沿成果，加速创新突破
获取完整内容-
Leica DMI6000B microscope
DMI6000B
Leica
Microscope for high-resolution imaging
暂无现货
预约到货通知
-
Yokogawa CSU-X1 spinning disk
CSU-X1
Yokogawa
Spinning disk unit for confocal microscopy
暂无现货
预约到货通知
-
Andor iXon camera
iXon
Andor
EMCCD camera for high-sensitivity imaging
暂无现货
预约到货通知
-
Andor FRAPPA unit
FRAPPA
Andor
Unit for photoactivation and photobleaching
暂无现货
预约到货通知
-
Andor Dragonfly spinning disk confocal system
Dragonfly
Andor
Confocal system for TIRF and SRRF-stream imaging
暂无现货
预约到货通知
-
CoolLED pE-4000 LED illumination system
pE-4000
CoolLED
LED illumination for photoactivation
立即获取同款设备 -
Andor Revolution spinning disk confocal system
Revolution
Andor
Imaging system for confocal microscopy
暂无现货
预约到货通知
-
Nikon Eclipse Ti2 inverted microscope
Eclipse Ti2
Nikon
Inverted microscope for TIRF imaging
暂无现货
预约到货通知
-
登录查看剩余6件设备及参数对照表
查看全部
