Uncoupling endosomal CLC chloride/proton exchange causes severe neurodegeneration

Abstract

CLC chloride/proton exchangers may support acidification of endolysosomes and raise their luminal Cl⁻ concentration. Disruption of endosomal ClC‐3 causes severe neurodegeneration. To assess the importance of ClC‐3 Cl⁻/H⁺ exchange, we now generate Clcn3^unc/unc mice in which ClC‐3 is converted into a Cl⁻ channel. Unlike Clcn3^−/− mice, Clcn3^unc/unc mice appear normal owing to compensation by ClC‐4 with which ClC‐3 forms heteromers. ClC‐4 protein levels are strongly reduced in Clcn3^−/−, but not in Clcn3^unc/unc mice because ClC‐3^unc binds and stabilizes ClC‐4 like wild‐type ClC‐3. Although mice lacking ClC‐4 appear healthy, its absence in Clcn3^unc/unc/Clcn4^−/− mice entails even stronger neurodegeneration than observed in Clcn3^−/− mice. A fraction of ClC‐3 is found on synaptic vesicles, but miniature postsynaptic currents and synaptic vesicle acidification are not affected in Clcn3^unc/unc or Clcn3^−/− mice before neurodegeneration sets in. Both, Cl⁻/H⁺‐exchange activity and the stabilizing effect on ClC‐4, are central to the biological function of ClC‐3.