title: Studies of lipopolysaccharide effects on the induction of α-synuclein pathology by exogenous fibrils in transgenic mice.
abstract: BACKGROUND
  Parkinson's disease (PD) is a progressive neurodegenerative disorder that is pathologically characterized by loss of dopaminergic neurons from the substantia nigra, the presence of aggregated α-synuclein (αS) and evidence of neuroinflammation. Experimental studies have shown that the cerebral injection of recombinant fibrillar αS, especially in αS transgenic mouse models, can induce the formation and spread of αS inclusion pathology. However, studies reporting this phenomenon did not consider the presence of lipopolysaccharide (LPS) in the injected αS, produced in E. coli, as a potential confound. The objectives of this study are to develop a method to remove the LPS contamination and investigate the differences in pathologies induced by αS containing LPS or αS highly purified of LPS.
  
  
  CONCLUSIONS
  We were able to remove >99.5% of the LPS contamination from the αS preparations through the addition of a cation exchange step during purification. The αS pathology induced by injection of fibrils produced from αS containing LPS or purified of LPS, showed a similar distribution pattern; however, there was less spread into the cortex of the mice injected with αS containing higher levels of LPS. As previously reported, injection of αS fibrils could induce astrogliosis, and αS inclusions were present within astrocytes in mice injected with fibrils comprised of αS with or without cation exchange purification. Furthermore, we identified the presence of αS pathology in ependymal cells in both groups of mice, which suggests the involvement of a novel mechanism for spread in this model of αS pathology.
authors.affiliation: Center for Translational Research in Neurodegenerative Disease, Department of Neuroscience, University of Florida, 1275 Center Drive, Room BMS J-483, PO Box 100159, Gainesville, FL, 32610, USA. njrutherford@ufl.edu.
authors.forename: Nicola J
authors.initials: NJ
authors.lastname: Rutherford
authors.affiliation: Center for Translational Research in Neurodegenerative Disease, Department of Neuroscience, University of Florida, 1275 Center Drive, Room BMS J-483, PO Box 100159, Gainesville, FL, 32610, USA. asacino@ufl.edu.
authors.forename: Amanda N
authors.initials: AN
authors.lastname: Sacino
authors.affiliation: Center for Translational Research in Neurodegenerative Disease, Department of Neuroscience, University of Florida, 1275 Center Drive, Room BMS J-483, PO Box 100159, Gainesville, FL, 32610, USA. mbrooks87@ufl.edu.
authors.forename: Mieu
authors.initials: M
authors.lastname: Brooks
authors.affiliation: Center for Translational Research in Neurodegenerative Disease, Department of Neuroscience, University of Florida, 1275 Center Drive, Room BMS J-483, PO Box 100159, Gainesville, FL, 32610, USA. caro.ceb81@ufl.edu.
authors.forename: Carolina
authors.initials: C
authors.lastname: Ceballos-Diaz
authors.affiliation: Center for Translational Research in Neurodegenerative Disease, Department of Neuroscience, University of Florida, 1275 Center Drive, Room BMS J-483, PO Box 100159, Gainesville, FL, 32610, USA. tomladd@ufl.edu.
authors.forename: Thomas B
authors.initials: TB
authors.lastname: Ladd
authors.affiliation: Center for Translational Research in Neurodegenerative Disease, Department of Neuroscience, University of Florida, 1275 Center Drive, Room BMS J-483, PO Box 100159, Gainesville, FL, 32610, USA. jasiehoward@ufl.edu.
authors.forename: Jasie K
authors.initials: JK
authors.lastname: Howard
authors.affiliation: Center for Translational Research in Neurodegenerative Disease, Department of Neuroscience, University of Florida, 1275 Center Drive, Room BMS J-483, PO Box 100159, Gainesville, FL, 32610, USA. tgolde@ufl.edu.
authors.forename: Todd E
authors.initials: TE
authors.lastname: Golde
authors.affiliation: Center for Translational Research in Neurodegenerative Disease, Department of Neuroscience, University of Florida, 1275 Center Drive, Room BMS J-483, PO Box 100159, Gainesville, FL, 32610, USA. bgiasson@ufl.edu.
authors.forename: Benoit I
authors.initials: BI
authors.lastname: Giasson
mesh_terms: D000818:Animals; D001253:Astrocytes / Q000473:pathology; D002452:Cell Count; D002478:Cells, Cultured; D002852:Chromatography, Ion Exchange; D018450:Disease Progression; D004340:Drug Contamination; D004731:Endotoxins / Q000302:isolation & purification / Q000494:pharmacology*; D018728:Entorhinal Cortex / Q000187:drug effects / Q000473:pathology; D004926:Escherichia coli / Q000737:chemistry / Q000378:metabolism; D006624:Hippocampus / Q000187:drug effects / Q000473:pathology; D006801:Humans; D057927:Hydrophobic and Hydrophilic Interactions; D002479:Inclusion Bodies / Q000737:chemistry; D007249:Inflammation; D007267:Injections; D051379:Mice; D008822:Mice, Transgenic; D017628:Microglia / Q000187:drug effects / Q000473:pathology; D020734:Parkinsonian Disorders / Q000139:chemically induced* / Q000473:pathology; D058225:Plaque, Amyloid / Q000737:chemistry; D011487:Protein Conformation; D011994:Recombinant Proteins / Q000302:isolation & purification / Q000494:pharmacology / Q000633:toxicity; D051844:alpha-Synuclein / Q000008:administration & dosage / Q000302:isolation & purification / Q000633:toxicity*
chemical_list: D004731:Endotoxins; D011994:Recombinant Proteins; D051844:alpha-Synuclein; C044101:endotoxin, Escherichia coli