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SRRF-Stream Imaging of Optogenetically Controlled Furrow Formation Shows Localized and Coordinated Endocytosis and Exocytosis Mediating Membrane Remodeling.
ACS Synthetic Biology ( IF 3.7 ) Pub Date : 2020-03-16 , DOI: 10.1021/acssynbio.9b00521
Jean A. Castillo-Badillo , Anoop C. Bandi , Suyash Harlalka , N. Gautam

Cleavage furrow formation during cytokinesis involves extensive membrane remodeling. In the absence of methods to exert dynamic control over these processes, it has been a challenge to examine the basis of this remodeling. Here we used a subcellular optogenetic approach to induce this at will and found that furrow formation is mediated by actomyosin contractility, retrograde plasma membrane flow, localized decrease in membrane tension, and endocytosis. FRAP, 4-D imaging, and inhibition or upregulation of endocytosis or exocytosis show that ARF6 and Exo70 dependent localized exocytosis supports a potential model for intercellular bridge elongation. TIRF and Super Resolution Radial Fluctuation (SRRF) stream microscopy show localized VAMP2-mediated exocytosis and incorporation of membrane lipids from vesicles into the plasma membrane at the front edge of the nascent daughter cell. Thus, spatially separated but coordinated plasma membrane depletion and addition are likely contributors to membrane remodeling during cytokinetic processes.

中文翻译:

SRRF流成像的光遗传学控制的沟形成显示局部和协调的内吞和胞吐介导膜重塑。

在胞质分裂过程中分裂沟的形成涉及广泛的膜重塑。在缺乏对这些过程进行动态控制的方法的情况下,检查这种重塑的基础是一个挑战。在这里,我们使用亚细胞光遗传学方法随意诱导这种现象,发现犁沟的形成是由放线菌素的收缩性,逆行的质膜流动,膜张力的局部降低和内吞作用介导的。FRAP,4-D成像以及内吞作用或胞吐作用的抑制或上调表明,ARF6和Exo70依赖性局部胞吐作用支持细胞间桥伸长的潜在模型。TIRF和超高分辨率径向波动(SRRF)流显微镜显示了局部VAMP2介导的胞吐作用,以及膜脂质从囊泡进入新生子细胞前缘的质膜。因此,在细胞动力学过程中,空间分离但协调的质膜消耗和增加可能是膜重塑的原因。
更新日期:2020-04-23
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