Protein‐bound crotonaldehyde accumulates in the spinal cord of superoxide dismutase‐1 mutation‐associated familial amyotrophic lateral sclerosis and its transgenic mouse model

Abstract

Growing evidence documents oxidative stress involvement in ALS. We previously demonstrated accumulation of a protein-bound form of the highly toxic lipid peroxidation product crotonaldehyde (CRA) in the spinal cord of sporadic ALS patients. In the present study, to the determine the role for CRA in the disease processes of superoxide dismutase-1 (SOD1) mutation-associated familial ALS (FALS), we performed immunohistochemical and semi-quantitative cell count analyses of protein-bound CRA (P-CRA) in the spinal cord of SOD1-mutated FALS and its transgenic mouse model. Immunohistochemical analysis revealed increased P-CRA immunoreactivity in the spinal cord of the FALS patients and the transgenic mice compared to their respective controls. In the FALS patients, P-CRA immunoreactivity was localized in almost all of the chromatolytic motor neurons, neurofilamentous conglomerates, spheroids, cordlike swollen axons, reactive astrocytes and microglia, and the surrounding neuropil in the affected areas represented by the anterior horns. In the transgenic mice, P-CRA immunoreactivity was localized in only a few ventral horn glia in the presymptomatic stage, in almost all of the vacuolated motor neurons and cordlike swollen axons and some of the ventral horn reactive astrocytes and microglia in the onset stage, and in many of the ventral horn reactive astrocytes and microglia in the advanced stage. Cell count analysis on mouse spinal cord sections disclosed a statistically significant increase in the density of P-CRA-immunoreactive glia in the ventral horns of the young to old G93A mice compared to the age-matched control mice. The present results indicate that enhanced CRA formation occurs in motor neurons and reactive glia in the spinal cord of SOD1-mutated FALS and its transgenic mouse model as well as sporadic ALS, sug- gesting implications for CRA in the pathomechanism common to these forms of ALS.

DJ-1 Changes in G93A-SOD1 Transgenic Mice: Implications for Oxidative Stress in ALS

“…Protein carbonylation measured immunologically revealed an almost exponential increase in these posttranslational modifications between 90 and 120 days in the SOD1 G93A mice . Lipid oxidation products are likewise increased in ALS mice spinal cords Perluigi et al 2005;Shibata et al 2007). Increased oxidative damage to DNA in spinal cord, cortex, and striatum of SOD1 G93A mice was reported as well (Aguirre et al 2005).…”

“…Multiple studies demonstrated increased oxidative damage to proteins, lipids, and DNA in ALS post mortem tissues (Barber et al 2006;Ilieva et al 2007). Evidence for oxidative damage has been found in the spinal cord and in the motor cortex from sporadic ALS cases (Shaw et al 1995;Fitzmaurice et al 1996;Ferrante et al 1997a;Shibata et al 2007) and SOD1 familial ALS patients (Abe et al 1995(Abe et al , 1997Beal et al 1997). Markers for protein and lipid oxidation were detected in motor neurons, reactive astrocytes, and microglia or macrophages (Shibata et al 2001).…”

Involvement of 4-hydroxy-2-nonenal Accumulation in Multiple System Atrophy

“…Immunohistochemical localization of P-HNE was verified by comparison with consecutive sections stained with H&E and immunostained with the antibodies to α-synuclein, ubiquitin, GFAP, and GLUT5, and it was also identified by the double-immunolabeled staining method as described previously [32]. Sections were incubated with one antibody, and immunoreaction was detected by the PIC method using DAB as the chromogen.…”

Aldehydes and Acetals

“…It can modify DNA by forming cyclic 1,N 2 -propanodeoxyguanosine adducts and can also induce micronuclei production and sister chromatid exchange (1,228). A potential role for crotonaldehydeprotein adducts in amyotrophic lateral sclerosis has been suggested (322). In mouse models of this disease, crotonaldehyde-immunoreactive neurons were identified and associated with the development of disease pathogenesis.…”