The subcellular arrangement of alpha-synuclein proteoforms in the Parkinson’s disease brain as revealed by multicolor STED microscopy

• DOI: 10.1007/s00401-021-02329-9
• Publication Date: 2021-06-11T10:03:15Z
• Authors: Tim E. Moors; Christina A. Maat; Daniel Niedieker; Daniel Mona; Dennis Petersen; Evelien Timmerman...; Jeroen Kole; Samir F. El-Mashtoly; Liz Spycher; Wagner Zago; Robin Barbour; Olaf Mundigl; Klaus Kaluza; Sylwia Huber; Melanie N. Hug; Thomas Kremer; Mirko Ritter; Sebastian Dziadek; Jeroen J. G. Geurts; Klaus Gerwert; Markus Britschgi; Wilma D. J. van d...
• Keywords: STED microscopy

Abstract Various post-translationally modified (PTM) proteoforms of alpha-synuclein (aSyn)—including C-terminally truncated (CTT) and Serine 129 phosphorylated (Ser129-p) aSyn—accumulate in Lewy bodies (LBs) different regions the Parkinson’s disease (PD) brain. Insight into distribution these within LBs subcellular compartments may aid understanding orchestration pathology PD. We applied epitope-specific antibodies against CTT Ser129-p aSyn domains immunohistochemical multiple labelings on post-mortem brain tissue from PD patients non-neurological, aged controls, which were scanned using high-resolution 3D multicolor confocal stimulated emission depletion (STED) microscopy. Our labeling setup highlighted a consistent onion skin-type architecture mature nigral an intricate structured-appearing framework cytoskeletal elements encapsulates core enriched species. By label-free CARS microscopy we found that enrichments proteins lipids mainly localized to central portion aSyn-immunopositive (aSyn+) inclusions. Outside LBs, observed 122CTT aSyn+ punctae at mitochondrial membranes cytoplasm neurons control brains, suggesting physiological role for outside LBs. In contrast, very limited no immunoreactivity was brains non-neurological while alignment neuronal cytoplasmic network characteristic with (incidental) LB disease. Interestingly, profiles not only containing but also without particularly donors early stage, pointing towards possible pathological phenotype preceding formation. Together, our observations human provide insights potential mechanisms underlying regulated morphogenesis.