The plasma membrane of Xenopus laevis oocytes contains voltage-dependent anion-selective porin channels.
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.
Steinacker
P
P
Max-Planck-Institut für Experimentelle Medizin, Abteilung Immunchemie, Göttingen, Germany.
Awni
L A
LA
Becker
S
S
Cole
T
T
Reymann
S
S
Hesse
D
D
Kratzin
H D
HD
Morris-Wortmann
C
C
Schwarzer
C
C
Thinnes
F P
FP
Hilschmann
N
N
D000595:Amino Acid Sequence; D000818:Animals; D000918:Antibody Specificity; D002462:Cell Membrane / Q000378:metabolism* / Q000648:ultrastructure; D005260:Female; D066298:In Vitro Techniques; D018613:Microscopy, Confocal; D008855:Microscopy, Electron, Scanning; D009865:Oocytes / Q000378:metabolism* / Q000648:ultrastructure; D018272:Porins / Q000235:genetics / Q000276:immunology / Q000378:metabolism*; D050994:Voltage-Dependent Anion Channels; D014982:Xenopus laevis
D018272:Porins; D050994:Voltage-Dependent Anion Channels