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Insulin-stimulated GLUT4 translocation is central to glucose homeostasis. Functional assays to distinguish individual steps in the GLUT4 translocation process are lacking, thus limiting progress toward elucidation of the underlying molecular mechanism. Here we have developed a robust method, which relies on dynamic tracking of single GLUT4 storage vesicles(More)
Large-conductance Ca(2+)-activated K(+) (BK) channels can regulate cellular excitability in complex ways because they are able to respond independently to two distinct cellular signals, cytosolic Ca(2+) and membrane potential. In rat chromaffin cells (RCC), inactivating BK(i) and noninactivating (BK(s)) channels differentially contribute to RCC action(More)
Feeding behaviour is modulated by both environmental cues and internal physiological states. Appetite is commonly boosted by the pleasant smell (or appearance) of food and destroyed by a bad taste. In reality, animals sense multiple environmental cues at the same time and it is not clear how these sensory inputs are integrated and a decision is made to(More)
GLUT4 is sequestered in intracellular storage compartments in a basal state and is rapidly translocated to the cell surface in response to insulin stimulation. Regulation of GLUT4 distribution is key for maintaining whole-body glucose homeostasis. To investigate the complicated intracellular movement of GLUT4 vesicles and their interactions with organelles(More)
Deconvolution wide-field fluorescence microscopy and single-particle tracking were used to study the three-dimensional mobility of single secretory granules in live PC12 cells. Acridine orange-labeled granules were found to travel primarily in random and caged diffusion, whereas only a small fraction of granules traveled in directed fashion. High K(+)(More)
We present a method for analysing propagation errors in membrane capacitance (C(m)) measurements under the whole-cell patch-clamp configuration, which mainly focusses on errors in C(m) estimates due to the 'residual' fast capacitance (DeltaC(p)). The method employs a quasi-phasor diagram for visualisation of the analysis. Our results show that both under-(More)
Computational modeling has emerged as an indispensable approach to resolve and predict the intricate interplay among the many ion channels underlying neuronal excitability. However, simulation results using the classic formula-based Hodgkin-Huxley (H-H) model or the superior Markov kinetic model of ion channels often deviate significantly from native(More)
Accurate Cm measurements rely on accurate determination of specific parameters of a patch-clamp amplifier (PCA). Hardware-related parameters, such as the resistance Rf and the stray capacitance Cf of the feedback resistor, the input capacitance Ci, the injection capacitance Cj, and the extra capacitances introduced by the BNC connector, are of significance(More)
In this paper, a programmable signal conditioning system based on software calibration for resistor-feedback patch clamp amplifier (PCA) has been described, this system is mainly composed of frequency correction, programmable gain and filter whose parameters are configured by software automatically to minimize the errors, A lab-designed data acquisition(More)
Membrane capacitance (C m) measurement, which measures and monitors plasma membrane capacitance and its changes of a cell, is one of the most important quantitative techniques and widely used in the studies of exocytosis and endocytosis. We present here a Z f-and-H sys-based C m measurement method (Z–H method) for our self-developed patch-clamp amplifier(More)