We have examined the oxidative metabolism of rat alpha-motoneurons innervating muscles composed predominantly of one muscle-fiber type. Intramuscular injections of horseradish peroxidase (HRP) into the tensor fasciae latae (TFL) (approximately 94% fast-twitch glycolytic fibers, FG), tibialis anterior (TA) (approximately 66% fast-twitch oxidative-glycolytic, FOG; 32% FG), and soleus (SOL) (approximately 84% slow-twitch oxidative, SO) muscles permitted identification of motoneurons innervating these muscles. gamma-Motoneurons (less than 25-micron average soma diameter) were eliminated from the sampling. The alpha-motoneurons innervating the TFL were considered as FG, those innervating the tibialis anterior as FOG, and those of the soleus as SO. Alternate 5-micron serial cryostat sections were processed for HRP and nicotinamide adenine dinucleotide-diapharase (NADH-D) (oxidative enzyme) activities. Controls were included to assure reliability of reaction product quantitation. Motoneuron pools of each muscle were characterized by their shape and location within the ventral horn. Cells identified on HRP sections as innervating each of the muscles were located on sections processed for NADH-D activity. The optical density of motoneurons in sections processed for NADH-D activity was measured with a Zeiss Zonax MPM 03 microdensitometer. The mean relative NADH-D activities (optical density) of alpha-motoneurons innervating the TFL (FG), TA (FOG), and SOL(SO) muscles were 0.261, 0.305, and 0.447, respectively. Although some overlap in distribution of enzyme activities was observed, statistical analysis indicated significant differences between the NADH-D activities of each type of alpha-motoneuron. This is the first report of any metabolic difference in alpha-motoneurons belonging to different motor-unit types.(ABSTRACT TRUNCATED AT 250 WORDS)