Relaxin is a peptide hormone found in seminal plasma that has a physiological influence on sperm motility in some species. There are no reports on the effect of relaxin on acrosome reaction and utilization of glucose in boar spermatozoa. In this study, to investigate the effects of relaxin on sperm motility, acrosome reaction, and incorporation and oxidation of labeled glucose, boar spermatozoa were washed and preincubated for swim-up and then incubated (0-6 h) with 0, 20, or 40 ng/ml relaxin in mTALP medium. The results indicated that the addition of relaxin stimulated sperm motility significantly (P<0.05) during 1-4 h of incubation. The percentage of acrosome-reacted live spermatozoa was higher (P<0.05) when the spermatozoa were treated with 20 or 40 ng/ml relaxin. The rate of incorporation, and oxidation of glucose were also greater (P<0.05) in the spermatozoa incubated with relaxin compared to the control spermatozoa. The rate of incorporation and oxidation of ¹⁴C-glucose were increased in correlation with acrosome reaction up to 4 h of incubation and then decreased in line with the increasing incubation period. In conclusion, the present study demonstrates that relaxin accelerates not only motility but also the acrosome reaction and utilization of glucose in boar spermatozoa.