Biodesign Center for Bioelectronics and Biosensors , Arizona State University , Tempe , Arizona 85287 , United States.
School of Electrical Computer and Energy Engineering , Arizona State University , Tempe , Arizona 85287 , United States.
ACS Sens. 2019 Aug 23;4(8):2205-2212. doi: 10.1021/acssensors.9b01201. Epub 2019 Aug 6.
Exocytosis involves interactions between secretory vesicles and the plasma membrane. Studying the membrane response is thus critical to understand this important cellular process and to differentiate different mediator release patterns. Here we introduce a label-free optical imaging method to detect the vesicle-membrane-interaction-induced membrane deformation associated with single exocytosis in mast cells. We show that the plasma membrane expands by a few tens of nanometers accompanying each vesicle-release event, but the dynamics of the membrane deformation varies from cell to cell, which reflect different exocytosis processes. Combining the temporal and spatial information allows us to resolve complex vesicle-release processes, such as two vesicle-release events that occur closely in time and location. Simultaneous following a vesicle release with fluorescence and membrane deformation tracking further allows us to determine the propagation speed of the vesicle-release-induced membrane deformation along the cell surface, which has an average value of 5.2 ± 1.8 μm/s.
胞吐作用涉及分泌囊泡与质膜之间的相互作用。因此,研究膜的反应对于理解这个重要的细胞过程和区分不同的介体释放模式至关重要。在这里,我们引入了一种无标记的光学成像方法来检测与肥大细胞中单胞吐作用相关的囊泡-膜相互作用引起的膜变形。我们发现,伴随每个囊泡释放事件,质膜会扩展几十纳米,但膜变形的动力学在细胞间存在差异,这反映了不同的胞吐作用过程。结合时间和空间信息,我们能够解析复杂的囊泡释放过程,例如在时间和位置上非常接近的两个囊泡释放事件。同时用荧光和膜变形跟踪来跟踪一个囊泡的释放,还使我们能够确定囊泡释放诱导的膜变形沿着细胞表面的传播速度,其平均值为 5.2 ± 1.8 μm/s。