Abstract
Oligodendroglial inclusion bodies characterize a subset of neurodegenerative diseases. Multiple system atrophy (MSA) is characterized by α-synuclein glial cytoplasmic inclusions and progressive supranuclear palsy (PSP) is associated with glial tau inclusions. The ubiquitin homologue, SUMO-1, has been identified in inclusion bodies in MSA, located in discrete sub-domains in α-synuclein-positive inclusions. We investigated SUMO-1 associated with oligodendroglial inclusion bodies in brain tissue from MSA and PSP and in glial cell models. We examined MSA and PSP cases and compared to age-matched normal controls. Fluorescence immunohistochemistry revealed frequent SUMO-1 sub-domains within and surrounding inclusions bodies in both diseases and showed punctate co-localization of SUMO-1 and the lysosomal marker, cathepsin D, in affected brain regions. Cell counting data revealed that 70–75 % of lysosomes in inclusion body-positive oligodendrocytes were SUMO-1-positive consistently across MSA and PSP cases, compared to 20 % in neighbouring inclusion body negative oligodendrocytes and 10 % in normal brain tissue. Hsp90 co-localized with some SUMO-1 puncta. We examined the SUMO-1 status of lysosomes in 1321N1 human glioma cells over-expressing α-synuclein and in immortalized rat oligodendrocyte cells over-expressing the four repeat form of tau following treatment with the proteasome inhibitor, MG132. We also transfected 1321N1 cells with the inherently aggregation-prone huntingtin exon 1 mutant, HttQ74-GFP. Each cell model showed the association of SUMO-1-positive lysosomes around focal cytoplasmic accumulations of α-synuclein, tau or HttQ74-GFP, respectively. Association of SUMO-1 with lysosomes was also detected in glial cells bearing α-synuclein aggregates in a rotenone-lesioned rat model. SUMO-1 labelling of lysosomes showed a major increase between 24 and 48 h post-incubation of 1321N1 cells with MG132 resulting in an increase in a 90 kDa SUMO-1-positive band that was immunopositive for Hsp90 and immunoprecipitated with an anti-SUMO-1 antibody. That SUMO-1 co-localizes with a subset of lysosomes in neurodegenerative diseases with glial protein aggregates and in glial cell culture models of protein aggregation suggests a role for SUMO-1 in lysosome function.
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Acknowledgments
We are grateful to the Australian Research Council, Griffith Health Institute and the Estate of the late Clem Jones AO for financial support, to P. Robinson for the gift of the anti-dynamin-1 anti-body, to R. De Silva for the gift of the anti-Tau antibody, to J. Yuan for the gift of the SUMO-1-eGFP transient transfection plasmid, to Dr. Andreas Wyttenbach for the HttQ74-eGFP transfection plasmid and to Poul Henning Jensen for the α-synuclein stable transfection plasmid. We thank Cobie Powell and Natalie Noe for assistance. Supported partially by the NHMRC 535014, 510186.
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Wong, M.B., Goodwin, J., Norazit, A. et al. SUMO-1 is Associated with a Subset of Lysosomes in Glial Protein Aggregate Diseases. Neurotox Res 23, 1–21 (2013). https://doi.org/10.1007/s12640-012-9358-z
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DOI: https://doi.org/10.1007/s12640-012-9358-z