The innervation of laryngeal muscle fibers was appraised in adult humans. Sixteen intrinsic laryngeal muscles were dissected during the autopsy of 4 adults (41-71 years old). Longitudinal serial frozen sections, 60 microm thick, of the whole muscles were double-stained for cholinesterase activity and axonal visualization. About 945 endplates per muscle were analysed using light microscopy. The neuromuscular junctions were always scattered throughout the whole muscles. Most of the muscle fibers showed a single neuromuscular junction, but multi-innervated fibers were found in all of the muscles. Their number was highest in interarytenoid muscles (21% of all the fibers). The distance between multiple neuromuscular junctions was most frequently less than 150 microm. Two neuromuscular junctions were frequently displayed, opposite one another, particularly in thyroarytenoid muscles, and this unusual feature seems specific for laryngeal muscles. The innervation of all of the muscle fibers was exclusively found to be unineuronal, with multi-innervated fibers being innervated by a single axon. Distal axonal degeneration occurred with aging, resulting in a loss in the number of multi-innervated muscle fibers.