Abstract Experiments were carried out examine short-term effects of human chorionic gonadotropin (hCG) and long-term effects of luteinising hormone releasing hormone analogue (LHRHa) on milt production of the tongue sole Cynoglossus semilaevis. In the first experiment, each fish was implanted with a blank cholesterol pellet (control), 100 and 200 μg LHRHa per kg body weight. In the second experiment, fish were injected with either 100, 200, 400 and 800 IU hCG per kg body weight or same volume of marine fish Ringer’s solution. In the first experiment, milt volume was increased in male implanted with 200 μg LHRHa pellet compared with other groups at day 10. Injection of 400 and 800 IU hCG resulted in an increase in the milt volume at hour 96 after the treatment. Although statistical difference is unable to confirm because of small milt volume, compared with the control group, hormone pellet-treated groups had a reduction in the mean spermatocrit (Sct) and sperm concentration (Sc). The results suggest that the increase in milt volume is at least partially gonadotropin (GtH)-dependent and increased milt volume has a relationship with milt hydration.Key words : Tongue sole, Cynoglossus semilaevis, spermiation, hormoneCorresponding author: 061-450-2395, limhk@mokpo.ac.kr2013 .