No abstract

The control of prostaglandin (PG) production by steroid hormones has been investigated in non-pregnant bilaterally ovariectomized sheep, prepared with indwelling utero-ovarian venous catheters and treated with physiological amounts of oestradiol and progesterone. Oestradiol treatment alone (2 \ m=x\ 15 \g=m\g/day for 9 days) had no effect on the prostaglandin F (PGF) concentration in uterine caruncles or intercaruncular tissue, on the release of PGF or of 13,14-dihydro-15-oxo PGF (PGFM) from these tissues during incubation in vitro, or on the concentrations of PGF in the utero-ovarian vein or PGFM in the jugular vein. However, oestradiol did accumulate in the uterine tissues. Progesterone treatment alone (2 \ m=x\10 mg/day for 9 days) provoked a significant increase in the concentration of PGF in the caruncles, a significant increase in the release of PGF from the caruncles during incubation with arachidonic acid and increased mean concentrations of PGFM in the jugular vein. When oestradiol was superimposed on a progesterone-primed system, there was a further marked increase in the PGF content of the caruncles, release of PGF into the utero-ovarian vein, and increased concentrations of PGFM in the jugular vein. The caruncles always contained more PGF than the intercaruncular area, and released more PGF and PGFM during incubation in vitro. In the progesterone+oestradiol group, there was good correlation between the PGF concentrations in simultaneous samples from the right and left utero-ovarian veins, and for all groups there was a high correlation between utero-ovarian PGF and peripheral PGFM concentrations. The caruncular epithelium of the progesterone-treated animals contained more lipid droplets than those of the other groups.These data are consistent with a requirement for progesterone in activating 'prostaglandin synthetase' activity, and promoting PGF production, largely from the caruncles. After progesterone priming, the synthesis of PGF by the caruncles and PG release into the vascular system was increased further by oestradiol treatment, whereas oestradiol alone was without effect.

The present work concerns the luteolytic effects of prostaglandin (PG) F2 alpha and its analogue, 16-aryloxy PGF2 alpha, upon isolated luteal cells. Varying doses of these two prostaglandins were incubated with cells in the presence or absence of an optimum stimulatory dose of LH (1 microgram/ml). The total contents of progesterone and 20 alpha-dihydroprogesterone in flasks were determined after the incubation periods by radioimmunoassay. Both prostaglandins inhibited basal synthesis of progesterone and 20 alpha-dihydroprogesterone, maximum inhibition occurring at concentrations of either PG of between 250 and 500 ng/ml. In this dose range both prostaglandins were found to abolish LH-stimulated progestogen synthesis completely. These effects were discernible within 5 min of incubation. The studies demonstrated that the onset of PG-induced luteolysis in vitro is characterized by an inhibition of the biosynthesis of both progesterone and its weakly progestogenic metabolite, 20 alpha-dihydroprogesterone; induction of 20 alpha-hydroxysteroid dehydrogenase activity by either PG was not found in incubations extending up to 60 min. In contrast to their relative potencies in vivo, PGF2 alpha and 16-aryloxy PGF2 alpha were essentially equipotent in this in-vitro system.

The acute antigonadotrophic action of prostaglandin F2 alpha (PGF2 alpha) was examined in dispersed luteal cell preparations from immature superluteinized rat ovaries. Cell suspensions prepared by collagenase digestion and purification over a Percoll density gradient were incubated for 1 h in Eagle's minimum essential medium in the presence and/or absence of LH, PGF2 alpha, N6,O2'-dibutyryladenosine 3',5'-cyclic monophosphate (dbcAMP) and forskolin. Medium was assayed for total progesterone and adenosine 3',5'-cyclic monophosphate (cAMP). Luteal cell preparations showed typical steroidogenic (progesterone) responses to LH, mimicked by both dbcAMP and forskolin. Whilst the threshold LH dose to increase cAMP synthesis was greater than that for progesterone (100 micrograms/l compared with 1 microgram/l), 24 mumol forskolin/l was the threshold dose for both cAMP and progesterone responses. Furthermore, combined doses of LH and forskolin synergistically raised cAMP yet produced less than additive increases in progesterone. Similarly, combinations of dbcAMP plus forskolin produced less than additive progesterone increases. These data suggest that forskolin may not act as a simple mimic of LH. Prostaglandin F2 alpha dose-dependently inhibited forskolin-induced cAMP and progesterone synthesis and also inhibited progesterone synthesis induced by dbcAMP. These data suggest that the antigonadotrophic effect of PGF2 alpha has more than one locus of action, i.e. it both inhibits an adenylate cyclase event associated with cAMP generation and blunts the cellular response to cAMP. The present uncertainty over the exact locus of forskolin's action within the adenylate cyclase complex limits further delineation of the inhibitory action of PGF2 alpha on LH-responsive adenylate cyclase.