This study investigated the effects of swimming training in different water temperatures on muscle GLUT4 protein expression and glycogen storage. Twenty-four female SD rats were weight-matched into three groups, including the control group (C), the 34℃ water temperature training group (W), and the 25℃ water temperature training group (CO). After two weeks of swimming training, the muscle GLUT4 protein gene expression, glycogen storage, and whole body glucose tolerance were measured. Our results showed that the glycogen content of red and white quadriceps muscles in CO and W groups were significantly higher than those of the control group. Meanwhile, the glycogen content of the red and white gastrocenimus muscles also revealed the consistent pattern as quadriceps muscles. Moreover, the GLUT4 protein expression of red and white gastrocenimus muscles in both warm-and cold-temperature exercise training groups were significantly higher than those in the control group. In conclusion, we found that swimming training in 25℃ water significantly enhanced glycogen storage but not the alteration in muscle GLUT4 protein expression.