14-3-3 proteins are highly conserved eukaryotic proteins that regulate various types of signal transduction pathways through phosphorylation-dependent protein-protein interactions. 14-3-3 mRNAs have been shown to be up-regulated in the injured rat motor neurons and in the spinal cords of patients with amyotrophic lateral sclerosis (ALS). To investigate the role of 14-3-3 proteins in ALS, we performed immunohistochemical studies on 14-3-3 using autopsied spinal cords from patients with sporadic ALS (sALS) and non-ALS subjects without spinal cord involvement. In the anterior horn of both groups, strong 14-3-3 immunoreactivity was observed in the somata and proximal processes of motor neurons. Many spheroids from all of the sALS cases were also immunopositive for 14-3-3. In addition, Lewy body-like hyaline inclusions (LBHIs), which were present in some sALS cases, were intensely immunostained. Our findings suggest that even in the severely affected anterior horn of patients with sALS, remaining motor neurons may contain abundant 14-3-3 proteins, and that 14-3-3 proteins may be partly associated with the pathogenesis of sALS, in particular with the formation of LBHIs.