Generation and regulation of the cell resting membrane potential - implications for function

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All cells have a resting membrane potential (RMP), due to the unequal ionic distribution across the membrane. Cellular integrity and function is closely coupled to the RMP with key processes including, for example, proliferation, secretion, and migration triggered by changes in membrane voltage. Hence, manipulation of the cell RMP has applications for health and disease, for example in cancer where cells have been shown to have RMPs that differ from their normal, healthy counterparts. The cell RMP is most readily recorded by microelectrode methods while specific fluorophores have been developed that measure ion concentrations within cells. Working with researchers equipped with expertise in electrophysiology, cell culture and live-cell fluorescence imaging, the aim of this rotation is to enable the student to sample these cutting-edge methodologies. The RMP in various cell lines will be measured in response to altered ionic gradients across the membrane to compare theoretical and experimental values. The dominant ionic species contributing to the cell RMP will also be determined using well-characterised ion channel blockers. A component of the training will involve the use of live-cell confocal imaging (at QMC) using known fluorophores that track potassium ions. Understanding key concepts of membrane biophysics will enable deeper mechanistic insight into thegeneration of the RMP and manipulation of cellular function.