Titlebook: Calcium Signalling; Methods and Protocol Anna Raffaello,Denis Vecellio Reane Book 2019 Springer Science+Business Media, LLC, part of Spring

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MCU Regulation in Lipid Bilayer and Electrophysiological Recording,l, using the electrophysiological technique of planar lipid bilayer..We showed that MCU gives rise to single-channel Ca. currents. In contrast, MCUb alone does not display calcium-permeable channel activity, while the co-expression of MCUb:MCU drastically alters the calcium permeation mediated by MCU subunit.

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Electrophysiological Characterization of Calcium-Permeable Channels Using Planar Lipid Bilayer,protein complex responsible for the uptake of this ion into mitochondria..Here we describe the protocol used for the electrophysiological characterization of the mitochondrial calcium uniporter (MCU) complex: the following outline indicates step-by-step the setup of planar lipid bilayer experiments.

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Imaging Intracellular Ca2+ in Cardiomyocytes with Genetically Encoded Fluorescent Probes,red contractility and detrimental remodeling of the cellular structure. For these reasons, the study of intracellular Ca. handling in cardiomyocytes represents a central method in experimental molecular cardiology.

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Employing Transgenic Parasite Strains to Study the Ca, Dynamics in the Human Malaria Parasite ,, . parasite. This method employs the construction of transgenic parasites (through standard molecular biology techniques), selection of the transfected population, and use of those parasites in spectrofluorometric Ca. assays.

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Assessing Calcium-Stimulated Mitochondrial Bioenergetics Using the Seahorse XF96 Analyzer,ical tips to design and perform controlled measurements of (a) respiratory and glycolytic metabolism of intact cells, (b) substrate-dependent respiration in permeabilized cells and isolated mitochondria, and (c) calcium-dependent regulation of mitochondrial bioenergetics with Seahorse XF Flux Analyzers.

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Textilforschungsanstalt Krefeldle alignment and image acquisition are detailed. Finally, the image processing steps to analyze calcium oscillations are discussed, with particular emphasis on ratiometric calcium imaging in . root hairs.