We examine the muscle fiber population of skeletal muscles from whole body in the cheetah (Acinonyx jubatus). In the present experiments, we showed the characteristics of fiber composition in the cheetah by comparative studies among the cheetah, domestic cat, and the beagle dog. Fiber population was determined on muscle fibers stained with monoclonal antibody to each myosin heavy chain isoform. Histochemical analysis demonstrated that many muscles in the cheetah and domestic cat had a low percentage of Type I fibers and a high percentage of Type IIx fibers, while those in the beagle dog showed a high percentage of Type IIa. The hindlimb muscles in the cheetah had a higher percentage of Type II (Type IIa + IIx) fiber than the forelimb muscles. This fact suggests that the propulsive role of the hindlimb is greater than the forelimb in the cheetah. The longissimus in the cheetah had a high percentage of Type IIx fibers over a wide range from the thoracic to lumbar parts, while the population of muscle fibers in this muscle was different depending on the parts in the domestic cat and beagle dog. This indicates that the cheetah can produce a strong and quick extension of the spinal column and increase its stiffness during locomotion. Furthermore, we found the notable difference of muscle fiber type population between flexors and extensors of digits in the cheetah. The present experiments show the characteristics of muscle fibers in the cheetah, corresponded to its ability to perform high-speed running.