Enhaced induction of the immunoproteasome by interferon gamma in neurons expressing mutant huntingtin

Abstract

Huntington disease (HD) inclusions are stained with anti-ubiquitin and anti-proteasome antibodies. This, together with proteasome activity studies on transfected cell, suggested that alterations in the ubiquitin-proteasome system (UPS) might contribute to HD pathogenesis. In a previous work we reported that in a conditional mouse model of Huntington’s disease (HD94 mice), the chymiotrypsin- and trypsin-“like” activities of the proteasome are increased selectively in the affected and aggregate-containing brain regions: striatum and cortex. Moreover, in these areas a neuronal increase in the interferon-inducible subunits of the immunoproteasome LMP2 and LPM7 was observed. In order to test if the expression ofN-terminal mutant huntingtin (htt) by itself is sufficient to induce the change in proteasome catalytic activities as well as in LMP2 subunit expression, we performed enzymatic assays for the three different peptidase activities of the proteasome and western blot experiments in striatal cultured neurons from HD94 mice free of glial contamination. We found no changes in any of the activities in these cells. Furthermore, western blot analysis performed with specific antibody against LMP2 subunits, revealed no difference in levels of this subunit in striatal neurons from HD94 compared to control cultures. On the other hand, when the striatal cultures were treated with interferon-γ (IFN-γ) during 72 hours, a clear increase in LMP2 levels was observed in control neuronal cultures. Interestingly, this increase was much more pronounced (95% higher) in HD94 striatal cultures. These results indicate that although expression of mutant htt is not sufficient to induce the changes in proteasome catalytic core observed in HD, it synergizes the changes induced by IFN-γ. Furthermore, immunocytochemical studies revealed that HD94 striatal neurons expressing high levels of LMP2 subunit showed a pre-apoptotic appearance. These results suggest that the correlation between neuronal induction of the inmuno-proteasome and neurodegeneration found in HD brains is secondary to inflammatory processes.