Proc Natl Acad Sci U S A 95(11): 6469-6473

α-Synuclein in filamentous inclusions of Lewy bodies from Parkinson’s disease and dementia with Lewy bodies

^{Medical Research Council Centre for Brain Repair and Department of Neurology, University of Cambridge, Robinson Way, Cambridge CB2 2PY, United Kingdom; and}^{Medical Research Council Laboratory of Molecular Biology, Hills Road, Cambridge CB2 2QH, United Kingdom}

^{To whom reprint requests should be addressed. e-mail:}ku.ca.mac.bml-crm@gm.

Communicated by Max F. Perutz, Medical Research Council, Cambridge, United Kingdom

Received 1998 Feb 18; Accepted 1998 Mar 25.

Abstract

Lewy bodies and Lewy neurites are the defining neuropathological characteristics of Parkinson’s disease and dementia with Lewy bodies. They are made of abnormal filamentous assemblies of unknown composition. We show here that Lewy bodies and Lewy neurites from Parkinson’s disease and dementia with Lewy bodies are stained strongly by antibodies directed against amino-terminal and carboxyl-terminal sequences of α-synuclein, showing the presence of full-length or close to full-length α-synuclein. The number of α-synuclein-stained structures exceeded that immunoreactive for ubiquitin, which is currently the most sensitive marker of Lewy bodies and Lewy neurites. Staining for α-synuclein thus will replace staining for ubiquitin as the preferred method for detecting Lewy bodies and Lewy neurites. We have isolated Lewy body filaments by a method used for the extraction of paired helical filaments from Alzheimer’s disease brain. By immunoelectron microscopy, extracted filaments were labeled strongly by anti-α-synuclein antibodies. The morphologies of the 5- to 10-nm filaments and their staining characteristics suggest that extended α-synuclein molecules run parallel to the filament axis and that the filaments are polar structures. These findings indicate that α-synuclein forms the major filamentous component of Lewy bodies and Lewy neurites.

Keywords: ubiquitin, sarkosyl-insoluble filaments, immunoelectron microscopy

Abstract

Parkinson’s disease (PD) is the second most common neurodegenerative disorder, after Alzheimer’s disease. Clinically, it is a movement disorder that is characterized by tremor, rigidity, and bradykinesia. Neuropathologically, PD is defined by nerve cell loss in the substantia nigra and the presence there of Lewy bodies and Lewy neurites (1, 2). Lewy bodies and Lewy neurites also are found in other brain regions, such as the dorsal motor nucleus of the vagus, the nucleus basalis of Meynert, and the locus coeruleus (reviewed in ref. 3). Brainstem-type Lewy bodies appear as intracytoplasmic inclusions, 5–25 μm in diameter, with a dense eosinophilic core and a clearer surrounding halo. Ultrastructurally, they are composed of a dense core of filamentous and granular material that is surrounded by radially oriented filaments (4, 5).

Abundant Lewy bodies and Lewy neurites in cerebral cortex are the defining neuropathological characteristics of dementia with Lewy bodies (DLB), a common late-life dementia that is clinically similar to Alzheimer’s disease (6–8). Cortical Lewy bodies are spherical intracytoplasmic inclusions that lack a distinctive core and halo, but that are made of filaments with morphologies similar to those of Lewy bodies from brainstem areas (6, 7).

Lewy neurites constitute an important part of the pathology of PD and DLB. They correspond to abnormal neurites that contain filaments similar to those found in Lewy bodies (9). A number of proteins have been identified immunologically by light microscopy in Lewy bodies and Lewy neurites from PD and DLB, which include neurofilaments and ubiquitin (10–15). Although these have been useful as markers for diagnostic purposes, the biochemical composition of Lewy body filaments has remained unknown.

Recently, the discovery of point mutations in the α-synuclein gene as a rare cause of familial PD (16, 17) has led us to the finding that α-synuclein is a component of Lewy bodies and Lewy neurites from idiopathic PD and DLB (18). We show here that antibodies raised against amino-terminal and carboxyl-terminal sequences of the 140-aa α-synuclein protein strongly immunostain Lewy bodies and Lewy neurites and that the number of stained structures exceeds that recognized by anti-ubiquitin antibodies. Antibodies directed against β-synuclein (19) and the BCSG1 protein (γ-synuclein) (20) failed to stain Lewy bodies and Lewy neurites. We also show that 5- to 10-nm filaments extracted from cingulate cortex of patients with DLB are labeled by anti-α-synuclein antibodies, indicating that α-synuclein is the major component of the filaments of Lewy bodies and Lewy neurites.

Acknowledgments

We thank Dr. N. J. Cairns and Professor P. L. Lantos (Medical Research Council Brain Bank, Department of Neuropathology, Institute of Psychiatry, London) and Dr. J. Xuereb (Medical Research Council Brain Bank, Department of Psychiatry, University of Cambridge, Cambridge, U.K.) for kindly providing the tissues from patients with Parkinson’s disease and dementia with Lewy bodies. We thank C. Villa for help with photography. M.H. is supported by a postdoctoral fellowship from Innogenetics Inc.

Acknowledgments

ABBREVIATIONS

PD	Parkinson’s disease
DLB	dementia with Lewy bodies

ABBREVIATIONS

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