The electromyographic properties of the cremaster muscle (CM) are quite different from other skeletal muscles. It shows excessive spontaneous discharges, and the motor unit shape and firing frequency of the CM muscle differ from that of limb muscles. In this study, CM of six adult cadavers and six orchiectomy specimens were used to reveal the detailed histology of the muscle and provide an anatomophysiological explanation for these unusual electromyographic properties. Routine histochemical stains revealed the CM was composed of several distinct bundles of smooth and striated muscle fibers within connective tissue. The smooth muscle fibers that were more profuse than previously known and were not arranged in layers, but widely dispersed between striated muscle fibers. Bielsehowsky silver staining technique, anti-neurofilament and anti-synaptophysin immunostaining showed the presence of multiple motor end-plates observed as a series of small dots or lines running along the striated muscle fibers and several nerve endings on a single muscle fiber. Myosin immunostaining confirmed the CM is a slow-twitch muscle, and a-actin smooth muscle immunostaining confirmed the presence of a large number of smooth muscle fibers. There were also small multipolar neurons forming nerve plexuses between smooth muscle fibers. Anti-GFAP immunostaining confirmed the presence of glial cells similar to astrocytes. In conclusion, the findings of this detailed anatomical study showed the CM, widely known as a striated muscle, contains a large number of smooth muscle fibers, and the spontaneous electromyographic discharges are due to the presence of multiple motor end-plates and dense innervation.