α-Synuclein Promotes the Recruitment of Tau to Protein Inclusions in Oligodendroglial Cells: Effects of Oxidative and Proteolytic Stress

Abstract

alpha-Synuclein is the major building block of cytoplasmic inclusions in neurodegenerative disorders named synucleinopathies. These inclusion bodies often contain the small heat shock protein alphaB-crystallin and the microtubule-associated protein tau. Oxidative modification of alpha-synuclein has been linked to fibril formation, and alpha-synuclein aggregation may induce the fibrillization of tau. To study alpha-synuclein aggregate formation, we have engineered oligodendroglial cells (OLN-93 cells) to stably express the longest human isoform of tau and wild-type alpha-synuclein or the A53T alpha-synuclein mutation. Under normal growth conditions, small punctuated alpha-synuclein aggregates were formed, which were more abundant in cells expressing the A53T mutation. After exposure to oxidative stress, protein inclusions were enlarged and were positive for thioflavin S, but the solubility of alpha-synuclein was not altered. Oxidative stress followed by proteasomal inhibition caused the occurrence of larger thioflavin S-positive inclusions, immunoreactive for tau and alphaB-crystallin, thus resembling glial cell inclusion bodies. Furthermore, this double stress situation led to a decrease in alpha-synuclein solubility, and alphaB-crystallin and HSP90 were present in the insoluble fraction. The formation and recruitment of tau to thioflavin S-positive protein aggregates in OLN-93 cells only expressing tau in the absence of alpha-synuclein, either after oxidative or proteasomal stress or both, was not observable. The data indicate that oxidatively modified alpha-synuclein is degraded by the proteasome and that it plays a pro-aggregatory role for tau in this cell culture model system.