title: Conformational templating of α-synuclein aggregates in neuronal-glial cultures.
abstract: BACKGROUND
  Genetic studies have established a causative role for α-synuclein (αS) in Parkinson's disease (PD), and the presence of αS aggregates in the form of Lewy body (LB) and Lewy neurite (LN) protein inclusions are defining pathological features of PD. Recent data has established that extracellular αS aggregates can induce intracellular αS pathologies supporting the hypothesis that αS pathology can spread via a "prion-like" self-templating mechanism.
  
  
  RESULTS
  Here we investigated the potential for conformational templating of αS intracellular aggregates by seeding using recombinant wild-type and PD-linked mutant (A53T and E46K) αS in primary mixed neuronal-glial cultures. We find that wild-type and A53T αS fibrils predominantly seed flame-like inclusions in both neurons and astrocytes of mixed primary cultures; whereas the structurally distinct E46K fibrils seed punctate, rounded inclusions. Notably, these differences in seeded inclusion formation in these cultures reflect differences in inclusion pathology seen in transgenic mice expressing the A53T or E46K αS mutants. We further show that the inclusion morphology is dictated primarily by the seed applied rather than the form of αS expressed. We also provide initial evidence that αS inclusion pathology can be passaged in primary astrocyte cultures.
  
  
  CONCLUSIONS
  These studies establish for the first time that αS aggregation in cultured cells can occur by a morphological self-templating mechanism.
authors.affiliation: Department of Neuroscience, McKnight Brain Institute, University of Florida College of Medicine, Gainesville, FL 32610, USA.
authors.forename: Amanda N
authors.initials: AN
authors.lastname: Sacino
authors.forename: Michael A
authors.initials: MA
authors.lastname: Thomas
authors.forename: Carolina
authors.initials: C
authors.lastname: Ceballos-Diaz
authors.forename: Pedro E
authors.initials: PE
authors.lastname: Cruz
authors.forename: Awilda M
authors.initials: AM
authors.lastname: Rosario
authors.forename: Jada
authors.initials: J
authors.lastname: Lewis
authors.forename: Benoit I
authors.initials: BI
authors.lastname: Giasson
authors.forename: Todd E
authors.initials: TE
authors.lastname: Golde
mesh_terms: D000818:Animals; D015153:Blotting, Western; D018929:Cell Culture Techniques; D018920:Coculture Techniques; D006801:Humans; D002479:Inclusion Bodies / Q000378:metabolism / Q000473:pathology*; D016631:Lewy Bodies / Q000378:metabolism / Q000473:pathology; D051379:Mice; D008856:Microscopy, Fluorescence; D009457:Neuroglia / Q000378:metabolism / Q000473:pathology*; D009474:Neurons / Q000378:metabolism / Q000473:pathology*; D010300:Parkinson Disease / Q000378:metabolism / Q000473:pathology; D051844:alpha-Synuclein / Q000378:metabolism*
chemical_list: D051844:alpha-Synuclein