Abstract
In Lewy body disease, alpha-synucleinopathic inclusions tend to develop in brain regions that extend long, thin, and unmyelinated projection fibers, but it is not known if this selective vulnerability is caused by poor myelination status. Our objective was to investigate this link in the limbic forebrain. First, we observed that levels of insoluble, hyperphosphorylated alpha-synuclein correlated inversely with the myelin marker proteolipid protein (20 kDa isoform) in the amygdala of men (but not women) with Lewy body disease. Second, select oligodendrocytic markers were mildly suppressed in the amygdala of men with Lewy body disease. Third, preformed fibrils (PFFs) of alpha-synuclein tended to induce greater pathology in gray matter, even when infused into murine brain regions penetrated by myelinated fiber bundles. Given these correlative patterns, we tested the hypothesis that myelin disruption with pharmacologic or genetic tools will exacerbate the development of limbic alpha-synucleinopathy in mice of both sexes, using a full-factorial experimental design. In nontransgenic mice with retrobulbar infusions of PFFs, dietary exposure to the established myelin disruptor cuprizone modestly increased the fraction of Triton-insoluble alpha-synuclein that was hyperphosphorylated at serine 129 (pSer129). Contrary to our hypothesis, however, cuprizone did not induce systematic, robust worsening of most pSer129+ object attributes, behavior deficits, cellular toxicity measures (NeuN+/Hoechst+ object attributes), or pan-alpha-synuclein insolubility. Similarly, PFF-induced effects on the insolubility and hyperphosphorylation of alpha-synuclein, behavior measures, and pSer129+, NeuN+, and Hoechst+ object attributes were not markedly amplified in mice heterozygous for the shiverer mutation in myelin basic protein (Mbp+/shi), compared to wild-type littermates. Although alpha-synucleinopathy is negatively associated with select myelin markers in mice and men, the causal nature of this link thus remains to be verified, and its potential implications are discussed here.

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Data availability
Full datasets for the bulk RNA-seq analyses are available at NCBI’s Gene Expression Omnibus (ID number GSE297212). The underlying code for this study is available on GitHub and can be accessed via this link: https://github.com/RachelClarkDUQ/RC_Cuprizone-Shiverer-Paper?tab=readme-ov-file#rc_cuprizone-shiverer-paper Other data will be made available upon request. Please email leakr@duq.edu.
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Acknowledgements
We are grateful for support by the NIH (1R15NS130532-01 and 1R21NS141002-01 to RKL). MA was supported by a Fulbright Scholarship from the Institute of International Education. We thank Walter Clark for outstanding assistance with the RStudio script. We are grateful to Drs. Johnnie Anderson and Torbjørn Kristensen at VisioPharm for their support with stereological measurements. We thank the Animal Care Facility (Rachel Barr, Ashley Yuhouse, Peyton Jackson, and Lindsay Hudson), the Center for Biologic Imaging at the University of Pittsburgh, the Graduate School of Pharmaceutical Sciences, and the Duquesne University Office of Research and Innovation for their kind support. We are grateful to Dr. Brian Popko for shipping us the shiverer breeders. JC is the Richard King Mellon Professor of Neurology at the University of Pittsburgh and supported by the VA Senior Research Career Scientist Award (821-RC-NB-30556) and Merit Review awards (I01BX005290 and I01BX003377) from the Department of Veterans Affairs.
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R.N.C. contributed to the methodology, software, validation, formal analysis, investigation, resources, visualization, and writing the original draft as well as review and editing. R.E.L. contributed to the methodology, formal analysis, investigation, writing the original draft as well as reviewing and editing the manuscript. M.A. and J.R.J contributed to methodology, investigation, and formal analysis. V.G.M., K.A.D., and J.F. contributed to formal analysis and investigation. J.C., K.C.L., and X.H. contributed to reviewing and editing the manuscript. R.K.L. contributed to the conceptualization, methodology, validation, formal analysis, investigation, resources, writing the original draft as well as review and editing, visualization, supervision, project administration, and funding acquisition.
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Clark, R.N., Landes, R.E., Abbas, M. et al. Testing an inverse link between limbic alpha-synucleinopathy and myelin markers in mice and humans. npj Parkinsons Dis. (2026). https://doi.org/10.1038/s41531-026-01278-y
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DOI: https://doi.org/10.1038/s41531-026-01278-y


