The authors developed a polyclonal antibody against a fusion protein containing 598 amino acids from a human choline acetyltransferase (ChAT) cDNA and 12 amino acids derived from an expression vector, and examined immunohistochemical reactivity for ChAT in large motor neurons (30 microns and more in somal minimal diameter) of the lumbar spinal cords of four patients with amyotrophic lateral sclerosis (ALS) and of four control cases. In controls, the number of large neurons included in the tissue with a total thickness of 100 microns ranged from 74 to 105 (average 87). About 60-90% (average 80%) of the neurons were positively stained in their perikarya with an anti-human ChAT antibody. In the cases of ALS, the number of large motor neurons was greatly reduced (25-60, average 38). About 4-13% (average 8%) were positively stained. These results indicate that not only large neurons are reduced in number, but also their positivity for ChAT is decreased in the anterior horn of ALS spinal cord.