The mylohyoid (MH) muscle plays a critical role in chewing, swallowing, respiration, and phonation. The present study was designed to test the hypothesis that the functional properties of the MH are reflected by its intrinsic specializations, including the neural organization, fiber-type distribution, and myosin heavy chain (MHC) expression. Adult human MH muscles were investigated to determine the nerve supply pattern using Sihler's stain, banding pattern and types of motor endplates using acetylcholinesterase (AChE) staining and silver impregnation, and muscle fiber type and MHC composition using immunocytochemical and immunoblotting techniques. The adult human MH was found to have the following neuromuscular specializations. First, the muscle was innervated by several branches of the MH nerve derived from the mandibular division of the trigeminal nerve. Each of the nerve branches supplied a distinct region of the muscle, forming a segmental innervation pattern. Second, the MH had a single motor endplate band which was located in the middle of the muscle length. Both en plaque and en grappe types of motor endplates were identified on the MH muscle fibers. Finally, the adult human MH fibers expressed unusual MHC isoforms (i.e., slow-tonic, alpha-cardiac, embryonic, and neonatal) which coexisted with the major MHC isoforms (i.e., slow type I, fast type IIa, and fast type IIx), thus forming various major/unusual (or m/u) MHC hybrid fiber types. The m/u hybrid fibers (84% of the total fiber population) were the predominant fiber types in the adult MH muscle. Determination of the neuromuscular specializations of the MH is helpful for better understanding of the muscle functions and for development of strategies to treat MH-related upper airway disorders.