Premium
Localization of LRRK2 to membranous and vesicular structures in mammalian brain
Author(s) -
Biskup Saskia,
Moore Darren J.,
Celsi Fulvio,
Higashi Shinji,
West Andrew B.,
Andrabi Shaida A.,
Kurkinen Kaisa,
Yu SeongWoon,
Savitt Joseph M.,
Waldvogel Henry J.,
Faull Richard L. M.,
Emson Piers C.,
Torp Reidun,
Ottersen Ole P.,
Dawson Ted M.,
Dawson Valina L.
Publication year - 2006
Publication title -
annals of neurology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 4.764
H-Index - 296
eISSN - 1531-8249
pISSN - 0364-5134
DOI - 10.1002/ana.21019
Subject(s) - lrrk2 , biology , microbiology and biotechnology , vesicular transport protein , endosome , organelle , synaptic vesicle , subcellular localization , colocalization , mitochondrion , cytosol , intracellular , vesicle , cytoplasm , biochemistry , membrane , gene , mutation , enzyme
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. Interpretation 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. Ann Neurol 2006;60:557–569