The effect of continuous human GH (hGH) secretion on female reproduction was studied in adult female transgenic rats expressing the hGH gene with a mouse whey acidic protein (mWAP) promoter. Two lines of transgenic female rats carrying the mWAP/hGH gene were established and used in the study. One was characterized by relatively high levels of serum hGH (high line), and the other had relatively low levels (low line). High-line female rats had recurring, pseudopregnancy-like estrous cycles accompanied by increased serum progesterone levels for 2 wk after ovulation, and they were fertile. In these rats, luteinization occurred spontaneously without cervical stimulation, probably due to high levels of serum hGH, which has prolactin (PRL)-like activity in the rat. Although low-line female rats had recurring, regular 4-day estrous cycles, they were sterile. In these rats, pseudopregnancy could not be induced by mating or by mechanical cervical stimulation. PRL surges following cervical stimulation were not detected, and PRL secretion was not induced by a dopamine antagonist, metoclopramide. The ovarian tissue in this line had a much higher number of corpora lutea and grew much heavier than in normal littermates, suggesting impairment of PRL-induced structural luteolysis. Suppression of PRL secretion in low-line rats was, at least in part, due to a marked decrease in the number of lactotrophs in the pituitary. The present study shows that the serum hGH level plays a crucial role in regulating luteal function in female transgenic rats expressing the hGH gene.