Hydroxynonenal adducts indicate a role for lipid peroxidation in neocortical and brainstem Lewy bodies in humans Rudy J. Castellani a , George Perry a, * , Sandra L. Siedlak a , Akihiko Nunomura b , Shun Shimohama c , Jing Zhang d , Thomas Montine d , Lawrence M. Sayre e , Mark A. Smith a a Institute of Pathology, Case Western Reserve University, 2085 Adelbert Road, Cleveland, OH 44106, USA b Department of Psychiatry and Neurology, Asahikawa Medical College, Asahikawa, Japan c Department of Neurology, Kyoto University, Kyoto, Japan d Department of Pathology, Vanderbilt University, Nashville, TN, USA e Department of Chemistry, Case Western Reserve University, Cleveland, OH, USA Received 8 October 2001; received in revised form 22 October 2001; accepted 14 November 2001 Abstract Multiple lines of evidence indicate that oxidative stress is a critical pathogenic factor in Parkinson disease (PD) and diffuse Lewy body disease (DLBD). Previously, we demonstrated increased levels of redox-active iron in Lewy bodies, and that Lewy bodies accumulate advanced glycation end-products. To further characterize the role of oxidative stress in diseases with Lewy body formation, we examined immunocytochemically eight cases of PD and five cases of DLBD for adducts of the lipid peroxidation adduct 4-hydroxy-2-nonenal, and for N 1 -(carboxymethyl)lysine (CML). Our findings demonstrate immunolocalization of 4-hydroxynonenal and CML to Lewy bodies in PD and DLBD. These findings not only support prior studies indicating that lipid peroxidation is increased in patients with PD and DLBD but that oxidative damage may play a critical role in Lewy body formation. q 2002 Elsevier Science Ireland Ltd. All rights reserved. Keywords: Diffuse Lewy body disease; Hydroxynonenal; Lewy body; N 1 -(carboxymethyl)lysine; Lipid peroxidation; Oxidative stress; Parkinson disease The central role of oxidative stress in Parkinson disease (PD) is indicated by a number of factors. Included among these is the presence of redox-active iron in pars compacta cells of the substantia nigra, diminished levels of antioxi- dants and antioxidant enzymes, oxidative metabolism of dopamine, immunodetection of advanced glycation end- products, nucleic acid and protein oxidation [1,2,20] and other markers of oxidative stress response. In Lewy bodies, advanced glycation [3] and protein nitration [7,13] have been specifically detected along with high levels of iron in the substantia nigra [10,18], and high levels of hydroxyl radical and plasma superoxide dismutase in PD patients compared with controls [10]. Recently, we have suggested that intraneuronal inclusion formation represents an adaptive mechanism for prolonging neuronal viability, based on the presence of a growing list of biochemical adaptations associated with oxidative stress that localize to these inclusions, and the empirical fact that such inclusions persist within viable neurons for years or decades [4,17]. To further characterize the mechanisms of oxidative stress associated with Lewy body formation, we studied cases of PD and diffuse Lewy body disease (DLBD) for evidence of lipid peroxidation, using antibodies to 4- hydroxy-2-nonenal (HNE) and N 1 -(carboxymethyl)lysine (CML). The latter is a late product of advanced glycation, but is also independently formed through lipid peroxidation, while the former is a lipid peroxidation adduct that is considered physiologically relevant to brain metabolism, and one of the most cytotoxic products of lipid peroxidation. Given the multiplicity of evidence and manifestations of oxidative stress in PD and DLBD, and the overlap in biochemical consequences, we hypothesized that lipid peroxidation is likely a critical component to this process, and that the cellular distribution of these processes may provide insight into Lewy body formation and, therefore, Neuroscience Letters 319 (2002) 25–28 0304-3940/02/$ - see front matter q 2002 Elsevier Science Ireland Ltd. All rights reserved. PII: S0304-3940(01)02514-9 www.elsevier.com/locate/neulet * Corresponding author. Tel.: 11-216-368-2488; fax: 11-216- 368-8964. E-mail address: gxp7@po.cwru.edu (G. Perry).