title: Localization of LRRK2 to membranous and vesicular structures in mammalian brain.
abstract: OBJECTIVE
  The PARK8 gene responsible for late-onset autosomal dominant Parkinson's disease encodes a large novel protein of unknown biological function termed leucine-rich repeat kinase 2 (LRRK2). The studies herein explore the localization of LRRK2 in the mammalian brain.
  
  
  METHODS
  Polyclonal antibodies generated against the amino or carboxy termini of LRRK2 were used to examine the biochemical, subcellular, and immunohistochemical distribution of LRRK2.
  
  
  RESULTS
  LRRK2 is detected in rat brain as an approximate 280kDa protein by Western blot analysis. Subcellular fractionation demonstrates the presence of LRRK2 in microsomal, synaptic vesicle-enriched and synaptosomal cytosolic fractions from rat brain, as well as the mitochondrial outer membrane. Immunohistochemical analysis of rat and human brain tissue and primary rat cortical neurons, with LRRK2-specific antibodies, shows widespread neuronal-specific labeling localized exclusively to punctate structures within perikarya, dendrites, and axons. Confocal colocalization analysis of primary cortical neurons shows partial yet significant overlap of LRRK2 immunoreactivity with markers specific for mitochondria and lysosomes. Furthermore, ultrastructural analysis in rodent basal ganglia detects LRRK2 immunoreactivity associated with membranous and vesicular intracellular structures, including lysosomes, endosomes, transport vesicles, and mitochondria.
  
  
  CONCLUSIONS
  The association of LRRK2 with a variety of membrane and vesicular structures, membrane-bound organelles, and microtubules suggests an affinity of LRRK2 for lipids or lipid-associated proteins and may suggest a potential role in the biogenesis and/or regulation of vesicular and membranous intracellular structures within the mammalian brain.
authors.affiliation: Institute for Cell Engineering, Johns Hopkins University School of Medicine, Baltimore, MD.|Department of Neurology, Johns Hopkins University School of Medicine, Baltimore, MD.
authors.forename: Saskia
authors.initials: S
authors.lastname: Biskup
authors.affiliation: Institute for Cell Engineering, Johns Hopkins University School of Medicine, Baltimore, MD.|Department of Neurology, Johns Hopkins University School of Medicine, Baltimore, MD.
authors.forename: Darren J
authors.initials: DJ
authors.lastname: Moore
authors.affiliation: Centre for Molecular Biology and Neuroscience, Institute of Basic Medical Sciences, University of Oslo, Oslo, Norway.
authors.forename: Fulvio
authors.initials: F
authors.lastname: Celsi
authors.affiliation: Department of Psychiatry, Juntendo University School of Medicine, Bungko-ku, Tokyo, Japan.|Neurobiology Programme, The Babraham Institute, Babraham, Cambridge, United Kingdom.
authors.forename: Shinji
authors.initials: S
authors.lastname: Higashi
authors.affiliation: Institute for Cell Engineering, Johns Hopkins University School of Medicine, Baltimore, MD.|Department of Neurology, Johns Hopkins University School of Medicine, Baltimore, MD.
authors.forename: Andrew B
authors.initials: AB
authors.lastname: West
authors.affiliation: Institute for Cell Engineering, Johns Hopkins University School of Medicine, Baltimore, MD.|Department of Neurology, Johns Hopkins University School of Medicine, Baltimore, MD.
authors.forename: Shaida A
authors.initials: SA
authors.lastname: Andrabi
authors.affiliation: Institute for Cell Engineering, Johns Hopkins University School of Medicine, Baltimore, MD.|Department of Neurology, Johns Hopkins University School of Medicine, Baltimore, MD.
authors.forename: Kaisa
authors.initials: K
authors.lastname: Kurkinen
authors.affiliation: Institute for Cell Engineering, Johns Hopkins University School of Medicine, Baltimore, MD.|Department of Neurology, Johns Hopkins University School of Medicine, Baltimore, MD.
authors.forename: Seong-Woon
authors.initials: SW
authors.lastname: Yu
authors.affiliation: Institute for Cell Engineering, Johns Hopkins University School of Medicine, Baltimore, MD.|Department of Neurology, Johns Hopkins University School of Medicine, Baltimore, MD.
authors.forename: Joseph M
authors.initials: JM
authors.lastname: Savitt
authors.affiliation: Department of Anatomy with Radiology, Faculty of Medical and Health Sciences, University of Auckland, Auckland, New Zealand.
authors.forename: Henry J
authors.initials: HJ
authors.lastname: Waldvogel
authors.affiliation: Department of Anatomy with Radiology, Faculty of Medical and Health Sciences, University of Auckland, Auckland, New Zealand.
authors.forename: Richard L M
authors.initials: RLM
authors.lastname: Faull
authors.affiliation: Neurobiology Programme, The Babraham Institute, Babraham, Cambridge, United Kingdom.
authors.forename: Piers C
authors.initials: PC
authors.lastname: Emson
authors.affiliation: Centre for Molecular Biology and Neuroscience, Institute of Basic Medical Sciences, University of Oslo, Oslo, Norway.
authors.forename: Reidun
authors.initials: R
authors.lastname: Torp
authors.affiliation: Centre for Molecular Biology and Neuroscience, Institute of Basic Medical Sciences, University of Oslo, Oslo, Norway.
authors.forename: Ole P
authors.initials: OP
authors.lastname: Ottersen
authors.affiliation: Institute for Cell Engineering, Johns Hopkins University School of Medicine, Baltimore, MD.|Department of Neurology, Johns Hopkins University School of Medicine, Baltimore, MD.|Department of Neuroscience, Johns Hopkins University School of Medicine, Baltimore, MD.
authors.forename: Ted M
authors.initials: TM
authors.lastname: Dawson
authors.affiliation: Institute for Cell Engineering, Johns Hopkins University School of Medicine, Baltimore, MD.|Department of Neurology, Johns Hopkins University School of Medicine, Baltimore, MD.|Department of Neuroscience, Johns Hopkins University School of Medicine, Baltimore, MD.|Department of Physiology, Johns Hopkins University School of Medicine, Baltimore, MD.
authors.forename: Valina L
authors.initials: VL
authors.lastname: Dawson
mesh_terms: D000818:Animals; D000915:Antibody Affinity; D001692:Biological Transport; D015153:Blotting, Western; D001921:Brain / Q000166:cytology / Q000378:metabolism*; D006801:Humans; D007150:Immunohistochemistry; D000071158:Leucine-Rich Repeat Serine-Threonine Protein Kinase-2; D051379:Mice; D009474:Neurons / Q000378:metabolism; D017346:Protein Serine-Threonine Kinases / Q000235:genetics* / Q000378:metabolism*; D050683:R-SNARE Proteins / Q000235:genetics* / Q000378:metabolism*; D051381:Rats; D013347:Subcellular Fractions / Q000378:metabolism
chemical_list: D050683:R-SNARE Proteins; D000071158:Leucine-Rich Repeat Serine-Threonine Protein Kinase-2; C495279:Lrrk2 protein, mouse; D017346:Protein Serine-Threonine Kinases