The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature.ĭifferent modes of recordings. Here, we provide detailed instructions on setting up PA and IPE, performing experiments and analyzing the data.Ĭatecholamine Cyclic voltammetry Cytosolic Electrochemistry Exocytosis Fusion pore IPE Patch amperometry Synaptic vesicle. In amperometric mode, IPE detects total catechols, whereas in cyclic voltammetric mode, it provides more specific information on the nature of the detected molecules and may selectively quantify catecholamines, providing a direct approach to determine cytosolic concentrations of catecholaminergic transmitters and their metabolites. IPE, on the other hand, measures cytosolic catecholamines that diffuse into the patch pipette following membrane rupture to achieve the whole-cell configuration. The method provides precise information on single-vesicle size and quantal content, fusion pore conductance, and permeability of the pore for catecholamines. PA, which is employed in cell-attached or excised inside-out patch clamp configuration, offers high-resolution patch capacitance measurements with simultaneous amperometric detection of catecholamines released during exocytosis. Both patch amperometry (PA) and intracellular patch electrochemistry (IPE) take advantage of a recording configuration where an electrochemical detector-carbon fiber electrode (CFE)-is housed inside a patch pipette.
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