Fetal viability does not affect the onset of stretch-induced labor and the increase in amniotic fluid prostaglandin F2 alpha and plasma prostaglandin F2 alpha metabolite levels.

The role of the fetus in the onset and progress of stretch-induced labor and in the change in amniotic fluid prostaglandin F2 alpha and plasma prostaglandin F2 alpha metabolite levels was evaluated in six normal pregnant women (group 1) and six women whose fetuses had been dead for more than one week (group 2). The uterus was distended by a balloon inflated with physiologic saline. Regular uterine contractions occurred, and increased in all patients. Within 21 hours, all patients delivered a normal baby in group 1 and a macerated fetus in group 2. There was no significant difference in induction-delivery interval between the two groups. Both groups showed a significant and similar range of increases in the levels of amniotic fluid prostaglandin F2 alpha and plasma prostaglandin F2 alpha metabolite during treatment (P less than 0.001). Thus, the fetus has no functional role in the onset and progress of stretch-induced labor or in the rise of amniotic fluid prostaglandin F2 alpha and plasma prostaglandin F2 alpha metabolite levels.     

Trypanosome-induced increase in prostaglandin F(2alpha) and its relationship with corpus luteum function in the goat

Plasma progesterone and 13,14-dihydro-15-keto Prostaglandin F(2alpha) (PGFM) were measured in normal (uninfected) and Trypanosoma congolense -infected adult goats for a period of 121 d, from May to August, during the breeding season in Kenya. Chronic trypanosomiasis rapidly increased the baseline plasma PGFM levels and the occurrence of irregular PGFM peaks in several infected goats. Progesterone luteal levels declined rapidly from the second and subsequent cycles post patency. Estrous cycles also became irregular but predominately shorter (8 to 19 d) before cessation from the second to fourth cycle following infection. The PGFM levels were still high during the acyclic period in all goats when progesterone levels were very low (1.4 to 2.4 nmol/l). The reciprocal increase in peripheral PGFM and decline in progesterone in these goats would suggest, in part, a trypanosome-induced PGF(2alpha) mediated luteolysis, and the possible involvement of prostaglandins in trypanosome-induced infertility in female goats.     

Matrix metalloproteinase expression and activity following prostaglandin F(2 alpha)-induced luteolysis

Luteal tissue contains matrix metalloproteinases (MMPs) that cleave specific components of the extracellular matrix (ECM) and are inhibited by tissue inhibitors of metalloproteinases (TIMPs). We previously reported a decrease in luteal TIMP-1 within 15 min of prostaglandin F(2 alpha) (PGF(2 alpha))-induced luteolysis. An increase in the MMP:TIMP ratio may promote ECM degradation and apoptosis, as observed in other tissues that undergo involution. The objectives of these experiments were to determine whether 1) PGF(2 alpha) affects expression of mRNA encoding fibrillar collagenases (MMP-1 and -13), gelatinases A and B (MMP-2 and -9), membrane type (mt)-1 MMP (MMP-14), stromelysin (MMP-3), and matrilysin (MMP-7), and 2) PGF(2 alpha) increases MMP activity during PGF(2 alpha)-induced luteolysis in sheep. Corpora lutea (n = 3-10/time point) were collected at 0, 15, and 30 min and 1, 2, 4, 6, 12, 24, and 48 h after PGF(2 alpha) administration. Northern blot analysis confirmed the presence of all MMPs except MMP-9. Expression of mRNA for the above MMPs (except MMP-2) increased significantly (P < 0.05) by 30 min, and all MMPs increased significantly (P < 0.05) by 6 h after PGF(2 alpha) administration. Expression of MMP-14 mRNA increased significantly (P < 0.05) by 15 min post-PGF(2 alpha) and remained elevated through 48 h. MMP activity in luteal homogenates (following proenzyme activation and inactivation of inhibitors) was increased significantly (P < 0.05) by 15 min and remained elevated through 48 h post-PGF(2 alpha). MMP activity was localized (in situ zymography) to the pericellular area of various cell types in the 0-h group and was markedly increased by 30 min post-PGF(2 alpha). MMP mRNA expression and activity were significantly increased following PGF(2 alpha) treatment. Increased MMP activity may promote ECM degradation during luteolysis.     

Route of prostaglandin F2alpha injection and luteolysis in mares (38519).

Nine groups of pony mares (3/group) were used in a 3 times 3 factorial experiment. The factors were dose of PGF-2 alpha (0, 0.25 of 1.25 mg and route of administration (im, iu or il). Mares were laparotomized and treated on day 7 postovulation. Jugular blood was collected for progesterone RIA at 0 (pretreatment) and 1,6,12,24,48, and 72 hr posttreatment. In mares given either 0.25 mg or 1.25 mg PGF-2alpha, progesterone concentrations were not significantly different among the three routes at any of the posttreatment times studied except at 6 hr posttreatment. In mares given 0.25 mg, progesterone concentrations at 6 hr was less (p less than 0.05) for mares injected im than for mares injected iu. Compared to pretreatment progesterone values, PGF2-alpha (0.25 mg and 1.25 mg groups combined) administration significantly decreased progesterone concentration by 12 hr posttreatment in mares injected im and 24 hr in mares injected iu or il. In the iu group, a significant increase in progesterone concentration occurred between 1 and 6 hr followed by a significant decrease at 12 hr posttreatment. There were no significant differences among the three routes for intervals from treatment to estrus or ovulation, length of posttreatment estrus or length of interovulatory interval. Injection of either 0.25 mg or 1.25 mg PGF-2alpha significantly shortened the interval from treatment to estrus. Although 0.25 mg tended to shorten the interval from treatment to ovulation and interovulatory interval, these two end points were significantly shortened only in mares given 1.25 mg PGF-2alpha. Results indicated that local administration (iu or il) did not improve the luteolytic efficacy of PGF-2alpha over systemic administration (im).     

Timing of prostaglandin F(2alpha) release episodes and oxytocin receptor development during luteolysis in the cow

The timing of PGF(2alpha) release and the timing and extent of the rise in endometrial oxytocin receptors was determined in relation to the timing of the progesterone fall during luteolysis in cycling cows. In cows undergoing luteolysis (n = 6), measurement of PGF(2alpha) metabolite in hourly plasma samples collected during daily 10 h sampling periods identified a total of 2.2+/-0.5 PGF(2alpha) release episodes per animal, each of 4.0+/-0.4 h duration. In cows in which luteolysis was not observed (n = 4) no PGF(2alpha) release episodes were identified. In a further three cows in which additional repeated uterine biopsies were collected on days 15, 17, 19, 21 and 23, endometrial oxytocin receptors were initially undetectable (<15 fmol/mg protein) but had increased to 120+/-19 fmol/mg protein prior to the initiation of PGF(2alpha) release episodes. Receptor concentrations then continued to increase reaching peak concentrations of 651+/-142 after luteolysis had been completed.     

Studies on the mechanism of action of prostaglandin F2 alpha induced luteolysis in rats

The effects of prostaglandin F2 alpha (PGF2 alpha) administration on the utilization of low density lipoprotein (LDL) and progesterone secretion were examined in dispersed luteal cells from rat ovaries. Immature rats were rendered pseudopregnant with administration of pregnant mare serum gonadotropin and human chorionic gonadotropin. Animals were sacrificed at different times after PGF2 alpha (5 mg/kg) or vehicle administration on day-5 of pseudopregnancy. Administration of PGF2 alpha in vivo decreased human chorionic gonadotropin (hCG) binding to luteal cell membranes in vitro but enhanced binding of LDL. Utilization of labelled cholesterol for steroid synthesis from reconstituted LDL [(3H)-CL-LDL] by dispersed luteal cells was enhanced following PGF2 alpha administration. This suggests that the LDL pathway is not suppressed during prostaglandin induced luteolysis. Progesterone and total progestin secretion in response to N6-2'-0-Dibutyryladenosine 3'5'-cyclic monophosphate (cAMP) was decreased at 2, 4 and 24 hours following PGF2 alpha administration demonstrating a post-cAMP defect in steroidogenesis. Addition of the hydroxylated sterols, 20 or 25-OH cholesterol as substrate stimulated progesterone secretion in vehicle treated rats in a dose dependent fashion with 20-OH cholesterol being more potent. Progesterone secretion in response to stimulation with luteinizing hormone (LH) and cAMP from vehicle treated rats was less than that observed with 20 or 25-OH cholesterol, indicating that endogenous substrate may be a limiting factor in steroid synthesis. The maximal capacity of luteal tissue to produce progestins following PGF2 alpha administration was determined with 20-OH cholesterol as the substrate. The results suggest that the post-cAMP defect at 4 hours following PGF2 alpha administration may be due to failure of the cells to mobilize endogenous cholesterol. However at 24 hours following PGF2 alpha administration the decreased ability of luteal cells to convert cholesterol to pregnenolone may contribute to decreased progesterone synthesis.     

Prostaglandin F2 alpha-induced prolactin release and luteolysis in the goat.

Injection of prostaglandin F2 alpha (PGF2 alpha) initiated a significant increase in plasma prolactin levels in all goats except those in anoestrus. Luteolysis occurred in non-pregnant goats during the mid luteal phase when the goats were given PGF2 alpha either with or without the suppression of prolactin release by bromocryptine (CB154). Luteolysis and subsequent parturition also occurred in pregnant goats in mid and late gestation after PGF2 alpha injection, with an associated release of prolactin and decrease in plasma progesterone. Acute prolactin release in response to injection of thyrotrophin releasing factor may have had a transient effect on plasma progesterone levels, but did not appear to be luteolytic in either pregnant or non-pregnant goats.     

Local effects of the sphingosine 1-phosphate on prostaglandin F2alpha-induced luteolysis in the pregnant rat

Since the regression of the corpus luteum (CL) occurs via a tightly controlled apoptotic process, studies were designed to determine if local administration of the antiapoptotic agent sphingosine 1-phosphate (S1P) effectively blocks the luteolytic action of prostaglandin F-2alpha (PGF-2α). On day 19 of pregnancy, 2 hr before systemic PGF-2α administration, rats were injected intrabursa with either S1P or vehicle (control). The activity of four caspases, which contribute to the initial (caspase-2, -8, and -9) and final (caspase-3) events in apoptosis was measured in pooled CL from four individual ovaries at 0 and 4 hr after PGF-2α injection. The expression of the phosphorylated form of AKT (pAKT) and tumor necrosis factor-alpha (TNF-α) was analyzed by ELISA. In addition, cell death was evaluated by electronic microscopy (EM) in CL 4 and 36 hr after PGF-2α injection. The activity of caspase-2, -3, and -8 was significantly greater by 4 hr after PGF-2α, but not caspase-9 activity. In contrast, expression of pAKT and TNF-α decreased significantly. Administration of S1P suppressed (P < 0.05) these effects, decreasing caspase activities and increasing pAKT and TNF-α expression. The administration of S1P also significantly decreased the percentage of luteal apoptotic cells induced by PGF-2α. PGF-2α treatment increased the prevalence of luteal cells with advanced signs of apoptosis (i.e., multiple nuclear fragments, chromatin condensation, or apoptotic bodies). S1P treatment suppressed these changes and increased the blood vessel density. These results suggest that S1P blocks the luteolytic effect of the PGF-2α by decreasing caspase-2, -3, and -8 activities and increasing AKT phosphorylation and TNF-α expression. Mol. Reprod. Dev. 76: 1153–1164, 2009. © 2009 Wiley-Liss, Inc.     

Influence of estradiol on the secretion of oxytocin and prostaglandin F2 alpha during luteolysis in the ewe.

Twenty ewes of mixed breeds were randomly assigned in equal numbers to one of four groups in a 2 x 2 factorial design. The factors were x-irradiation to destroy ovarian follicles or sham irradiation and the administration of estradiol-containing or empty (placebo) implants. Surgery for irradiation was performed on Day 8 of the cycle. Blood samples were withdrawn from jugular catheters at 1.5-h intervals from Day 10 to Day 17. Luteolysis was not observed by Day 17 in 4 of 5 placebo-treated ewes after destruction of ovarian follicles. Luteolysis was observed in 4 of 5 ewes of the sham-irradiated, placebo-treated group and in all ewes that received estradiol whether or not ovarian follicles had been destroyed. The longest (p less than 0.07) interval between peaks of 13,14-dihydro-15-keto-prostaglandin F2 alpha (PGFM) was observed in the x-irradiated, placebo-treated group, whereas the administration of estradiol reduced (p less than 0.01) the interval between PGFM peaks. These findings indicate that a short interpulse interval in the secretion of prostaglandin F2 alpha (PGF2 alpha) is associated with luteolysis. It is possible that the reduced interpulse interval was either an effect of estradiol that caused luteolysis or a secondary event resulting from luteolysis. The administration of estradiol decreased (p less than 0.05) the number of episodes of oxytocin secretion during luteolysis and increased (p less than 0.01) the interval between episodes.(ABSTRACT TRUNCATED AT 250 WORDS)     

Suppression of prostaglandin F-2 alpha release and delay of luteolysis after active immunization against oxytocin in the goat

Active immunization against oxytocin significantly prolonged the oestrous cycle in 3 out of 4 goats; the mean (+/- s.e.m.) cycle length was 29.1 +/- 1.7 days (n = 12) compared to 19.4 +/- 0.6 days (n = 9) in control animals. During Days 10-21 of the cycle in the 3 responsive goats, peripheral plasma concentrations of progesterone and oxytocin were steady and those of 13,14-dihydro-15-keto-prostaglandin F-2 alpha were very low (50-100 pg X ml-1) with no marked pulsatile activity. The major effect of immunization would appear to be suppression of the synthesis of the uterine luteolysin PGF-2 alpha, thus confirming that endogenous oxytocin has a facilitatory role in luteolysis via prostaglandin production.     

Trilostane but not prostaglandin F2alpha (PGF2alpha) or cortisol aborts 90-day-pregnant lutectomized sheep

Ewes were lutectomized and treatments were started 72 h later. Pregnant ewes were treated with vehicle; prostaglandin F2alpha (PGF2alpha); cortisol (C); trilostane (TR), a 3beta-hydroxy-steroid dehydrogenase inhibitor; PGF2alpha + C; TR + PGF2alpha; TR + C, or TR + PGF2 + C. TR, TR + PGF2alpha, TR + C, and TR + PGF2alpha + C aborted (P < or = 0.05) all ewes receiving TR. One ewe treated with PGF2alpha aborted (P > or = 0.05). The average time to abortion of TR-treated ewes was 50.8 h (P < or = 0.05) after initiation of treatments. All aborted ewes had retained placentas (P < or = 0.05) except one ewe in the TR + PGF2alpha, treatment group. TR was given every 12 h starting at 72 h postlutectomy until 96 h postlutectomy. TR reduced (P < or = 0.05) progesterone. Estradiol-17beta was increased (P < or = 0.05) 2 h after the first two TR treatments and declined 2 h later and was followed by a sustained increase (P < or = 0.05) in estradiol-17beta, which was coincident with the onset of abortions. Estradiol-17beta was increased (P < or = 0.05) by PGF2alpha but did not decrease (P > or = 0.05) placental secretion of progesterone. It is concluded that TR but not PGF2alpha is an abortifacient in 90-day-pregnant lutectomized ewes and that abortion occurs only when there is a decrease in circulating progesterone and an increase in circulating estradiol-17beta.     

The role of luteal oxytocin in episodic secretion of prostaglandin F2alpha at luteolysis in the ewe

The role of luteal oxytocin in the generation of luteolytic episodes of prostaglandin F2alpha at luteolysis was investigated. On day 10 of the cycle Dorset ewes underwent either surgical removal of the corpora lutea (lutectomy; n = 4) or sham operation (sham; n = 4). Lutectomised ewes were then administered progesterone by twice daily i.m. injection in corn oil (20 mg/day) until day 14 when treatment was ceased to simulate luteolysis. The concentration of 13, 14 dihydro-15-keto prostaglandin F2alpha (PGFM) was measured in peripheral blood samples collected at 20-min intervals for 8 h on days 12-16 of the cycle. Progesterone and oestradiol concentrations were similar in the two groups over the whole experimental cycle while oxytocin fell dramatically following lutectomy. No prostaglandin F2alpha release episodes were seen on day 12 or 13, while from days 14-16 both groups exhibited a similar episode frequency (lutectomy 0.9/ewe/8 h; sham 0.8/ewe/8 h). Analysis of episode characteristics revealed lower episode amplitude (p<0.05) but longer episode duration (p<0.05) in the lutectomy group. The results demonstrate that a normal frequency of prostaglandin F2alpha release episodes occurs independently of luteal oxytocin secretion. However, luteal oxytocin is involved in regulating the pattern of release, perhaps causing the release of episodes of the magnitude required for the successful completion of luteolysis.     

Canine prostaglandin F2alpha receptor (FP) and prostaglandin F2alpha synthase (PGFS): molecular cloning and expression in the corpus luteum

In the dog luteolysis is not affected by hysterectomy. This observation led to the hypothesis that paracrine/autocrine rather than endocrine mechanisms of PGF2alpha are responsible for luteal regression in the dioestric bitch. The present experiments tested for the capacity of canine CL to produce and respond to PGF2alpha by qualitatively and quantitatively determining the expressions of PGFS, the enzyme converting PGH2 into PGF2alpha, and the PGF2alpha-receptor (FP) in CL of non-pregnant dogs during dioestrus. Canine PGFS and FP were isolated and cloned; both genes show a high homology (82-94%) when compared to those of other species. Relatively weak FP mRNA expression was detected on day 5 of dioestrus. It had increased by day 25 and remained constant thereafter. In situ hybridization (ISH) localized FP solely to the cytoplasm of the luteal cells, suggesting that these cells are the only luteal targets of PGF2alpha in this species. Only negative results were obtained for the expression of PGFS in canine CL by routine qualitative RT-PCR. When Real Time (TaqMan) PCR was applied, repetitively more negative than positive results were obtained at all timepoints. Any positive measurements observed at any point were neither repeatable nor related to the stage of dioestrus. This led us to conclude that expression of PGFS is either absent or present at very low level only. These data suggest that luteal regression in non-pregnant bitches is not modulated by PGF2alpha. However, the FP seems to be constitutionally expressed, explaining the receptivity of canine CL to exogenous PGF2alpha.     

Effect of dose and time of injection of prostaglandin F(2alpha) in cycling ewes

A study was done to evaluate the efficacy of graded doses of prostaglandin F(2alpha) (PGF(2alpha)) to induce regression of the corpus luteum and hence estrus, in cycling ewes when given on various days of the estrous cycle. One hundred cycling cross-bred ewes were observed twice daily (08:00 and 20:00 h) for marking by raddled vasectomized rams. After estrus was confirmed in marked ewes by assay of plasma progesterone concentration, the ewes were treated in pairs with 0, 5, 10, 15 or 20 mg PGF(2alpha) on day 2, 3, 4, 7, 8, 9, 12, 13, 14 or 15 of an estrous cycle and then exposed to a raddled ram of known libido and fertility. Plasma progesterone levels were determined on the day of, and on the day following PGF(2alpha)-treatment to monitor luteal function. Ewes marked between one and five days after treatment and having a decrease in plasma progesterone were considered to have responded to the treatment. The percentages of ewes responding were 10, 35, 60, 70 and 95 to doses of 0, 5, 10, 15 and 20 mg PGF(2alpha) respectively. Differences due to dose were significant (P < 0.01) with the two higher doses being more effective. There were differences due to the day of injection, with treatments on days 2 and 3 being less effective.     

Failure of the prostaglandin F2 alpha analogue, cloprostenol, to induce functional luteolysis in the olive baboon (Papio cynocephalus anubis)

The prostaglandin F2 alpha analogue, cloprostenol, which is an effective luteolytic agent in the common marmoset, was administered intramuscularly to olive baboons to determine if it possessed luteolytic properties in this species. The results showed that functional luteolysis was not induced when cloprostenol was administered during the mid- to late luteal phase or during early pregnancy.     

Coupling between cyclooxygenases and prostaglandin F(2alpha) synthase. Detection of an inducible,glutathione-activated,membrane-bound prostaglandin F(2alpha)-synthetic activity

Distinct functional coupling between cyclooxygenases (COXs) and specific terminal prostanoid synthases leads to phase-specific production of particular prostaglandins (PGs). In this study, we examined the coupling between COX isozymes and PGF synthase (PGFS). Co-transfection of COXs with PGFS-I belonging to the aldo-keto reductase family into HEK293 cells resulted in increased production of PGF(2alpha) only when a high concentration of exogenous arachidonic acid (AA) was supplied. However, this enzyme failed to produce PGF(2alpha) from endogenous AA, even though significant increase in PGF(2alpha) production occurred in cells transfected with COX-2 alone. This poor COX/PGFS-I coupling was likely to arise from their distinct subcellular localization. Measurement of PGF(2alpha)-synthetic enzyme activity in homogenates of several cells revealed another type of PGFS activity that was membrane-bound, glutathione (GSH)-activated, and stimulus-inducible. In vivo, membrane-bound PGFS activity was elevated in the lung of lipopolysaccharide-treated mice. Taken together, our results suggest the presence of a novel, membrane-associated form of PGFS that is stimulus-inducible and is likely to be preferentially coupled with COX-2.     

Boar spermatozoa and prostaglandin F2alpha. Quality of boar sperm after the addition of prostaglandin F2alpha to the short-term extender over cooling time

Prostaglandin F(2alpha) (PGF(2alpha)) has been used to improve reproductive performance in swine. The goal of the present work was to determine how the addition of PGF(2alpha) affects boar sperm quality. Eleven different treatments were evaluated: eight with only PGF(2alpha) (0.625, 1.25, 2.50, 5, 10, 12.50, 25 and 50mg PGF(2alpha)/100ml) and three binary treatments (0.625mg PGF(2alpha)/100ml+200mug/ml hyaluronic acid (HA), 1.25mg PGF(2alpha)/100ml+200mug/ml HA, 0.625mg PGF(2alpha)/100ml+7.5muM caffeine (Caf)). All these substances were added to 16 ejaculates from 16 healthy and sexually mature boars (n=16), and each ejaculate was considered as a replicate. Our study also assessed the effects of these 11 treatments over different periods of preservation. Sperm quality was tested immediately after the addition of treatments (time 0), and after 1, 3, 6 and 10 days of cooling at 15 degrees C. To evaluate sperm quality, five parameters were analysed: (1) sperm viability, acrosome and mitochondrial sheath integrity (using a multiple fluorochrome-staining test), (2) sperm motility, (3) sperm morphology and (4) agglutination (using a computer assisted system) and (5) osmotic resistance (using the ORT). Parametric (analysis of variance for repeated measures) and non-parametric tests (Friedman test) were used as statistical analyses. Treatments with PGF(2alpha) concentrations higher than 12.5mg/100ml were cytotoxic while the others did not damage boar spermatozoa. Thus, the other treatments may be used to produce profitable effects without adverse effects. Moreover, the addition of PGF(2alpha) at 5mg/100ml to sperm diluted in BTS may maintain sperm viability and motility better after 6 days of cooling, because significant differences were observed (P<0.05) compared with control at the same time.     

Concentrations of 13, 14-dihydro-15-keto prostaglandin F2 alpha after pulsatile progesterone administration at the time of luteolysis of heifers

Progesterone was administered in pulses to 12 dairy heifers from days 17.5 to 22.5 post-estrus in order to determine its ability to modify secretion of PGF2 alpha around the time of luteolysis. Control heifers exhibited pulses of PGFM concomitant with a sharp decline in progesterone concentrations and thus these pulses were temporally associated with luteolysis. Additional pulses of PGFM were observed in heifers receiving exogenous progesterone, but these were not statistically predictable by either dose of progesterone (50 or 100 micrograms) or time of administration (3 or 6 hour intervals). However, all heifers (4/4) treated with progesterone at 3 hour intervals had additional pulses of PGFM as compared to only one heifer (1/4) treated at 6 hour intervals. When pulses of PGFM were induced by exogenous progesterone there was a substantial lag time between the initiation of progesterone treatment and their occurrence. The limited response to progesterone administration and the lack of synchrony is not consistent with an ability of exogenous progesterone to directly stimulate secretion of PGF2 alpha at the time of luteolysis.     

Analysis of luteal tissue inhibitor of metalloproteinase-1, -2, and -3 during prostaglandin F(2alpha)-induced luteolysis

Increased matrix metalloproteinase (MMP) expression and activities help to mediate tissue involution through increasing extracellular matrix remodeling and promoting dedifferentiation and, ultimately, apoptosis. Therefore, we hypothesized that prostaglandin (PG) F(2alpha) administration would decrease expression of the tissue inhibitor of metalloproteinase (TIMP)-1, -2, and -3 and effectively increase the MMP:TIMP ratio, leading to glandular involution. In experiment 1, we tested the effects of PGF(2alpha) administration (Day 10 postestrus; Day 0 = estrus) on luteal TIMP-1, -2, and -3 mRNA and protein expression. Corpora lutea were collected at 0, 15, or 30 min or at 1, 2, 4, 6, 12, 24, and 48 h following PGF(2alpha) administration (n = 3-9 animals/time point). Following PGF(2alpha) administration, TIMP-1 mRNA levels decreased (P < 0.05) at 1 and 2 h relative to 0 h (controls), then increased to levels greater than controls at 4 and 6 h. In contrast, TIMP-2 and -3 mRNA levels did not decrease following PGF(2alpha) administration. The TIMP-1, -2, and -3 proteins were localized to large luteal cells (LLCs) within control (untreated) tissues. However, histodepletion of TIMP-1 within LLCs was evident within 30 min (earliest time point collected) following PGF(2alpha) injection and continued through 48 h. Luteal concentration of TIMP-1, as determined by RIA, was decreased (P < 0.05) by 15 min (earliest time point collected) following PGF(2alpha) administration and remained low through 48 h. In contrast, TIMP-2 and -3 immunolocalization was not altered by PGF(2alpha) administration. Experiment 2 was conducted to determine if PGF(2alpha) could initiate the preceding changes in TIMP-1 in early (Day 3) corpora lutea that can bind PGF(2alpha) but are refractory to its luteolytic effects. Serum concentrations of progesterone and luteal concentrations of TIMP-1 mRNA and protein were similar at 0 and 6 h after PGF(2alpha) injection on Day 3 postestrus. These data suggest that an early and sustained effect of PGF(2alpha) is the specific depletion of TIMP-1 within LLCs that are capable of responding to the luteolytic action of PGF(2alpha). This action may increase the MMP:TIMP-1 ratio, creating an environment that favors extracellular matrix degradation and, thereby, facilitates both functional and structural regression.