Measurement of dynamic membrane mechanosensation using optical tweezers.

Many cellular processes are orchestrated by dynamic changes in the plasma membrane to form membrane projections and endocytic vesicles in response to extracellular environmental changes. Our previous studies show that ARF6-ACAP4-ezrin signaling regulates membrane dynamics and curvature in response to EGF stimulation. However, there is no quantitative measurement to relate molecular organization of membrane cytoskeletal remodeling to stimulus-elicited mechanosensation on the plasma membrane. Optical tweezers is a powerful tool in the study of membrane tension. Comparing to pulling out an entire membrane tether at one time, the step-like method is more efficient because multiple relaxation curves can be obtained from one membrane tether. Fewer models describe relaxation curves to characterize mechanical properties of cell membrane. Here we establish a new method to measure the membrane relaxation curve of HeLa cells judged by the relationship between membrane tether diameter and tensions. We obtained effective viscosities and static tensions by fitting relaxation curves to our model. We noticed the delicate structure of relaxation curves contains information of cytoskeletal remodeling and lateral protein diffusion. Our study established a quantitative measure to characterize the mechanosensation of epithelial cells in response to stimulus-elicited membrane dynamics.