This preliminary study was designed to determine whether the neurotransmitter substance P was present in the middle carpal synovial membrane of the normal horse and whether the neuropeptide could be identified in the synovial fluid of normal horses and those with joint diseases. Immunocytochemistry on middle carpal synovial membrane biopsies from fresh cadavers was used to demonstrate substance P-containing neural elements. Substance P was most abundant in the subintimal portion of the membrane, with occasional filaments coursing via synovial fronds to the intimal portion. Radioimmunoassay techniques were used on acidified acetonitrile-preserved synovial fluid samples to measure substance P concentrations. Fluid from 9 joints of 5 normal horses and 6 joints of 4 horses with joint diseases were analysed. Disease conditions included acute and chronic osteoarthritis and osteochondrosis. Synovia from normal horses contained a mean concentration of substance P significantly less than that of horses with joint diseases (P less than 0.05). Elevated concentrations of neurotransmitters in diseased joints suggests a potential contribution to the pathophysiology of joint disorders in horses.