S.B. and D.J.M. contributed equally to this work.
Localization of LRRK2 to membranous and vesicular structures in mammalian brain
Article first published online: 21 NOV 2006
Copyright © 2006 American Neurological Association
Annals of Neurology
Volume 60, Issue 5, pages 557–569, November 2006
How to Cite
Biskup, S., Moore, D. J., Celsi, F., Higashi, S., West, A. B., Andrabi, S. A., Kurkinen, K., Yu, S.-W., Savitt, J. M., Waldvogel, H. J., Faull, R. L. M., Emson, P. C., Torp, R., Ottersen, O. P., Dawson, T. M. and Dawson, V. L. (2006), Localization of LRRK2 to membranous and vesicular structures in mammalian brain. Ann Neurol., 60: 557–569. doi: 10.1002/ana.21019
- Issue published online: 21 NOV 2006
- Article first published online: 21 NOV 2006
- Manuscript Accepted: 15 SEP 2006
- Manuscript Revised: 11 SEP 2006
- Manuscript Received: 29 MAR 2006
- NIH (National Institute of Neurological Disorders and Stroke [NINDS]). Grant Numbers: NS 38377, F32 NS053264
- Lee Martin Trust
- Sylvia Nachlas Trust
- National Parkinson Foundation
- American Parkinson Disease Association
- German Research Foundation (Deutsche Forschungsgemeinschaft)
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.
Polyclonal antibodies generated against the amino or carboxy termini of LRRK2 were used to examine the biochemical, subcellular, and immunohistochemical distribution of LRRK2.
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.
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