Localization of LRRK2 to membranous and vesicular structures in mammalian brain

Authors

  • Saskia Biskup MD, PhD,

    1. Institute for Cell Engineering, Johns Hopkins University School of Medicine, Baltimore, MD
    2. Department of Neurology, Johns Hopkins University School of Medicine, Baltimore, MD
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    • S.B. and D.J.M. contributed equally to this work.

  • Darren J. Moore PhD,

    1. Institute for Cell Engineering, Johns Hopkins University School of Medicine, Baltimore, MD
    2. Department of Neurology, Johns Hopkins University School of Medicine, Baltimore, MD
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    • S.B. and D.J.M. contributed equally to this work.

  • Fulvio Celsi BS,

    1. Centre for Molecular Biology and Neuroscience, Institute of Basic Medical Sciences, University of Oslo, Oslo, Norway
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  • Shinji Higashi MD,

    1. Department of Psychiatry, Juntendo University School of Medicine, Bungko-ku, Tokyo, Japan
    2. Neurobiology Programme, The Babraham Institute, Babraham, Cambridge, United Kingdom
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  • Andrew B. West PhD,

    1. Institute for Cell Engineering, Johns Hopkins University School of Medicine, Baltimore, MD
    2. Department of Neurology, Johns Hopkins University School of Medicine, Baltimore, MD
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  • Shaida A. Andrabi PhD,

    1. Institute for Cell Engineering, Johns Hopkins University School of Medicine, Baltimore, MD
    2. Department of Neurology, Johns Hopkins University School of Medicine, Baltimore, MD
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  • Kaisa Kurkinen PhD,

    1. Institute for Cell Engineering, Johns Hopkins University School of Medicine, Baltimore, MD
    2. Department of Neurology, Johns Hopkins University School of Medicine, Baltimore, MD
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  • Seong-Woon Yu PhD,

    1. Institute for Cell Engineering, Johns Hopkins University School of Medicine, Baltimore, MD
    2. Department of Neurology, Johns Hopkins University School of Medicine, Baltimore, MD
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  • Joseph M. Savitt MD, PhD,

    1. Institute for Cell Engineering, Johns Hopkins University School of Medicine, Baltimore, MD
    2. Department of Neurology, Johns Hopkins University School of Medicine, Baltimore, MD
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  • Henry J. Waldvogel PhD,

    1. Department of Anatomy with Radiology, Faculty of Medical and Health Sciences, University of Auckland, Auckland, New Zealand
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  • Richard L. M. Faull MD, PhD,

    1. Department of Anatomy with Radiology, Faculty of Medical and Health Sciences, University of Auckland, Auckland, New Zealand
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  • Piers C. Emson PhD,

    1. Neurobiology Programme, The Babraham Institute, Babraham, Cambridge, United Kingdom
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  • Reidun Torp PhD,

    1. Centre for Molecular Biology and Neuroscience, Institute of Basic Medical Sciences, University of Oslo, Oslo, Norway
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  • Ole P. Ottersen MD, PhD,

    1. Centre for Molecular Biology and Neuroscience, Institute of Basic Medical Sciences, University of Oslo, Oslo, Norway
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  • Ted M. Dawson MD, PhD,

    1. Institute for Cell Engineering, Johns Hopkins University School of Medicine, Baltimore, MD
    2. Department of Neurology, Johns Hopkins University School of Medicine, Baltimore, MD
    3. Department of Neuroscience, Johns Hopkins University School of Medicine, Baltimore, MD
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  • Valina L. Dawson PhD

    Corresponding author
    1. Institute for Cell Engineering, Johns Hopkins University School of Medicine, Baltimore, MD
    2. Department of Neurology, Johns Hopkins University School of Medicine, Baltimore, MD
    3. Department of Neuroscience, Johns Hopkins University School of Medicine, Baltimore, MD
    4. Department of Physiology, Johns Hopkins University School of Medicine, Baltimore, MD
    • Johns Hopkins University School of Medicine, 733 North Broadway, Suite 731, Broadway Research Building, Baltimore, MD 21205
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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.

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

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