Analysis of large conductance Ca2+-activated K+ (BK) channels dynamics in living cells.

Large conductance Ca2+-activated K+ (BK) channels are expressed in a wide range of human tissues such as brain, muscle, lung or pancreas, where they regulate insulin secretion by closing N-type calcium channels. The study of BK channels activity, localization and dynamics, would be useful not only for the achievement of a further understanding of BK channels properties in physiological conditions, but also in disease. Indeed, BK channel activity has been reported to be disrupted in a wide range of disorders, such as diabetes. Therefore, our study aims to determine whether components of the secretory machinery and BK channels are localised in close proximity at the plasma membrane, as well as how the secretory machinery influences BK channel localisation, activity and dynamics. To carry out these experiments we are using electrophysiology (patch-clamp) and super-resolution microscopy (gated-Stimulated Emission Depletion [g-STED], Photoactivatable Localisation Microscopy [PALM], single particle tracking PALM [sptPALM], Fluorescence Lifetime Imaging and Fluorescence Resonance Energy Transfer [FLIM-FRET]).