Treatment of ewes with steroid-free ovine follicular fluid (oFF) during the follicular phase of the oestrous cycle results in the immediate inhibition of the ovarian secretion of oestradiol, inhibin and androgens. An experiment was conducted to determine whether this effect of oFF was due to inhibin, or to direct inhibition of ovarian function by other factors in oFF. Eight ewes in which the left ovary and vascular pedicle had been autotransplanted to a site in the neck were studied during the breeding season. Luteal regression was induced in all animals by injection of cloprostenol (100 micrograms i.m.; PG) on Day 10 of the luteal phase. The animals were divided into two groups (n = 4) and treated with either steroid-free oFF (oFF; 3 ml s.c.; 3.2 microgram p1-26 alpha inhibin/ml) or steroid-free oFF in which the inhibin content had been reduced by greater than 90% (IFoFF; 3 ml s.c.; 0.3 microgram p1-26 alpha inhibin/ml) by affinity chromatography, 24 and 36 h after PG. Samples of ovarian and jugular venous blood were collected at (i) intervals of 4 h from 16 h before until 120 h after PG and (ii) intervals of 10 min from 48 to 52 h after injection of PG to investigate the pattern of pulsatile secretion of ovarian hormones. All ewes had previously been monitored during a normal PG-induced follicular phase. Injection of oFF resulted in an increase (P less than 0.05) in the concentration of inhibin in jugular venous plasma and a profound (P less than 0.001) and prolonged decrease in the peripheral concentration of follicle-stimulating hormone (FSH). Injection of IFoFF had no significant effect on peripheral concentrations of inhibin or FSH in the first 24 h after treatment; thereafter inhibin concentrations fell (P less than 0.01) progressively until 40 h and then increased (P less than 0.01) until 72 h after treatment. In both treatment groups, however, within 24-36 h of treatment the concentration of FSH increased 5-10-fold (P less than 0.001) to a peak that occurred within 48-60 h and then declined to basal concentrations within 72-84 h of treatment. The concentration of luteinizing hormone (LH) in jugular venous plasma increased in both groups after treatment (P less than 0.01), although the rise after injection of oFF only started after 24 h. Thereafter, there was a progressive increase in the concentration of LH, peaks occurring 48-60 h after treatment.(ABSTRACT TRUNCATED AT 400 WORDS)