The volume of milt that could be stripped from male goldfish, Carassius auratus, increased dramatically when fish were exposed overnight to water with concentrations of 17α,20β-dihydroxy-4-pregnen-3-one (17,20P) as low as 10−10 M. A variety of free steroids (pregnenolone, androstenedione, testosterone, 11-ketotestosterone, 17β-estradiol) and glucuronated steroids (etiocholanolone glucuronide, testosterone glucuronide, 17β-estradiol glucuronide), suggested by others to function as pheromones in fish, failed to increase milt volume at a concentration of 5 × 10−9 M. The milt response appears to be specific to 17,20P and progesterone precursors (17α-hydroxyprogesterone, progesterone), but is most sensitive to 17.20P. Because bilateral section of the olfactory tracts completely blocked the milt response to 17.20P, it is believed to be a pheromone. Selective sectioning of the lateral and medial subdivisions of the olfactory tracts demonstrated that the milt response to 17,20P is dependent on the medial tract. Increases in milt in response to 17,20P are not affected by interaction with conspecifics; exposure to 17,20P induced equivalent responses in males held in isolation, paired with a male, or paired with a female. It is proposed that 17,20P, which is produced during oocyte final maturation in the female, is a "primer" pheromone released to the water and that it increases milt volume in the male. This effect is discussed in relation to releaser pheromone effects previously demonstrated in goldfish.