Expression of human pICln and ClC-6 in Xenopus oocytes induces an identical endogenous chloride conductance.
pICln is a protein that induces an outwardly rectifying, nucleotide-sensitive chloride current (ICln) when expressed in Xenopus oocytes, but its precise function (plasma-membrane anion channel versus cytosolic regulator of a channel) remains controversial. We now report that a chloride current identical to ICln is induced when Xenopus oocytes are injected with human ClC-6 RNA. Indeed, both the pICln and the ClC-6 induced current are outwardly rectifying, they inactivate slowly at positive potentials and have an anion permeability sequence NO3- > I- > Br- > Cl- > gluconate. Cyclamate, NPPB, and extracellular cAMP block the induced currents. The success rate of current expression is significantly increased when the injected Xenopus oocytes are incubated at a higher temperature (24 or 37 degrees C) prior to the analysis. In addition, the ICln current was detected in 6.2% of noninjected control Xenopus oocytes. We therefore conclude that the ICln current in Xenopus oocytes corresponds to an endogenous conductance that can be activated by expression of structurally unrelated proteins. Furthermore, functional, biochemical, and morphological observations did not support the notion that pICln resides in the plasma membrane either permanently or transiently after cell swelling. Thus, it is unlikely that pICln forms the channel that is responsible for the ICln current in Xenopus oocytes.
Buyse
G
G
Laboratory of Physiology, Catholic University of Leuven, Campus Gasthuisberg, B-3000 Leuven, Belgium.
Voets
T
T
Tytgat
J
J
De Greef
C
C
Droogmans
G
G
Nilius
B
B
Eggermont
J
J
D000818:Animals; D015153:Blotting, Western; D018118:Chloride Channels / Q000378:metabolism*; D005260:Female; D019084:Fluorescent Antibody Technique, Indirect; D006801:Humans; D009865:Oocytes / Q000378:metabolism*; D016133:Polymerase Chain Reaction; D013347:Subcellular Fractions / Q000737:chemistry; D013696:Temperature; D014981:Xenopus
C097539:CLCN6 protein, human; D018118:Chloride Channels