Localization of LRRK2 to membranous and vesicular structures in mammalian brain.
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.
Biskup
Saskia
S
Institute for Cell Engineering, Johns Hopkins University School of Medicine, Baltimore, MD.|Department of Neurology, Johns Hopkins University School of Medicine, Baltimore, MD.
Moore
Darren J
DJ
Institute for Cell Engineering, Johns Hopkins University School of Medicine, Baltimore, MD.|Department of Neurology, Johns Hopkins University School of Medicine, Baltimore, MD.
Celsi
Fulvio
F
Centre for Molecular Biology and Neuroscience, Institute of Basic Medical Sciences, University of Oslo, Oslo, Norway.
Higashi
Shinji
S
Department of Psychiatry, Juntendo University School of Medicine, Bungko-ku, Tokyo, Japan.|Neurobiology Programme, The Babraham Institute, Babraham, Cambridge, United Kingdom.
West
Andrew B
AB
Institute for Cell Engineering, Johns Hopkins University School of Medicine, Baltimore, MD.|Department of Neurology, Johns Hopkins University School of Medicine, Baltimore, MD.
Andrabi
Shaida A
SA
Institute for Cell Engineering, Johns Hopkins University School of Medicine, Baltimore, MD.|Department of Neurology, Johns Hopkins University School of Medicine, Baltimore, MD.
Kurkinen
Kaisa
K
Institute for Cell Engineering, Johns Hopkins University School of Medicine, Baltimore, MD.|Department of Neurology, Johns Hopkins University School of Medicine, Baltimore, MD.
Yu
Seong-Woon
SW
Institute for Cell Engineering, Johns Hopkins University School of Medicine, Baltimore, MD.|Department of Neurology, Johns Hopkins University School of Medicine, Baltimore, MD.
Savitt
Joseph M
JM
Institute for Cell Engineering, Johns Hopkins University School of Medicine, Baltimore, MD.|Department of Neurology, Johns Hopkins University School of Medicine, Baltimore, MD.
Waldvogel
Henry J
HJ
Department of Anatomy with Radiology, Faculty of Medical and Health Sciences, University of Auckland, Auckland, New Zealand.
Faull
Richard L M
RLM
Department of Anatomy with Radiology, Faculty of Medical and Health Sciences, University of Auckland, Auckland, New Zealand.
Emson
Piers C
PC
Neurobiology Programme, The Babraham Institute, Babraham, Cambridge, United Kingdom.
Torp
Reidun
R
Centre for Molecular Biology and Neuroscience, Institute of Basic Medical Sciences, University of Oslo, Oslo, Norway.
Ottersen
Ole P
OP
Centre for Molecular Biology and Neuroscience, Institute of Basic Medical Sciences, University of Oslo, Oslo, Norway.
Dawson
Ted M
TM
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.
Dawson
Valina L
VL
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.
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
D050683:R-SNARE Proteins; D000071158:Leucine-Rich Repeat Serine-Threonine Protein Kinase-2; C495279:Lrrk2 protein, mouse; D017346:Protein Serine-Threonine Kinases