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The plasma membrane of Xenopus laevis oocytes contains voltage-dependent anion-selective porin channels
Authors:Steinacker P  Awni L A  Becker S  Cole T  Reymann S  Hesse D  Kratzin H D  Morris-Wortmann C  Schwarzer C  Thinnes F P  Hilschmann N
Institution:1. Institute of Parasitology, Biology Centre ASCR, České Budějovice CZ-370 05, Czech Republic;2. Institute of Entomology, Biology Centre ASCR, České Budějovice CZ-370 05, Czech Republic;1. Provincial Key Laboratory for Botanical Pesticide R&D of Shaanxi Province, Yangling, Shaanxi 712100, PR China;2. Institute of Pesticide Science, College of Plant Protection, Northwest A&F University, Yangling, Shaanxi 712100, PR China;3. Key Laboratory of Crop Pest Integrated Management on the Loess Plateau, Ministry of Agriculture, Yangling, Shaanxi 712100, PR China;4. Department of Entomology, Michigan State University, East Lansing, MI 48824, USA
Abstract:Recent patch-clamp studies have shown that anti-porin antibodies, applied to the external side of excised plasma membrane patches of mammalian astrocytes, close chloride channels that are thought to be engaged in cell volume regulation. Frog oocytes are often used to study this basic cell function. Here we document the localisation of endogenous porin voltage-dependent anion-selective channels in Xenopus laevis oocyte plasma membranes. In confocal laser microscopy images a disjunctive pattern of fluorescing spots appear about 10 microm apart. Labelling was prevented by preabsorption of the antibodies with synthetic peptides comprising the epitope of the antigen. Immuno-gold marking of oocyte surfaces followed by silver enhancement of the gold particles lead to a plasma membrane labelling corresponding to that obtained by the confocal laser approach. The data suggests the presence of voltage-dependent, anion-selective channels in oocyte plasma membranes. This data should be borne in mind when frog oocytes are used to study the characteristics of endogenous or heterologously expressed ion channels or regulatory proteins.
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