The Victor Chang Cardiac Research Institute, Lowy Packer Building, 405 Liverpool St, Darlinghurst, NSW 2010, Australia.
School of Medical Sciences, University of New South Wales, Kensington, NSW 2052, Australia.
Sci Rep. 2017 Mar 27;7:45180. doi: 10.1038/srep45180.
The droplet on hydrogel bilayer (DHB) is a novel platform for investigating the function of ion channels. Advantages of this setup include tight control of all bilayer components, which is compelling for the investigation of mechanosensitive (MS) ion channels, since they are highly sensitive to their lipid environment. However, the activation of MS ion channels in planar supported lipid bilayers, such as the DHB, has not yet been established. Here we present the activation of the large conductance MS channel of E. coli, (MscL), in DHBs. By selectively stretching the droplet monolayer with nanolitre injections of buffer, we induced quantifiable DHB tension, which could be related to channel activity. The MscL activity response revealed that the droplet monolayer tension equilibrated over time, likely by insertion of lipid from solution. Our study thus establishes a method to controllably activate MS channels in DHBs and thereby advances studies of MS channels in this novel platform.
水凝胶双层（DHB）上的液滴是研究离子通道功能的新型平台。该设置的优点包括对所有双层成分的严格控制，这对于研究机械敏感（MS）离子通道非常有吸引力，因为它们对其脂质环境非常敏感。然而，在平面支撑脂质双层（如 DHB）中，MS 离子通道的激活尚未得到证实。在这里，我们展示了 DHB 中大肠杆菌大电导 MS 通道（MscL）的激活。通过用纳升缓冲液选择性地拉伸液滴单层，我们诱导了可量化的 DHB 张力，这与通道活性有关。MscL 活性反应表明，液滴单层张力随时间平衡，可能是通过从溶液中插入脂质来实现的。因此，我们的研究建立了一种在 DHB 中可控激活 MS 通道的方法，从而推进了该新型平台中 MS 通道的研究。
