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Corpus luteum maintenance during early pregnancy in the cow.

Holstein heifers (18 pregnant, 12 inseminated but non-pregnant, and 17 cyclic) were studied during the first 18 days after oestrus. Jugular vein plasma progesterone concentrations were significantly (P < 0.05) higher in pregnant than in non-pregnant and cyclic animals between Days 10 and 18. Jugular oestradiol concentrations were also higher in pregnant than in cyclic animals between Days 6 and 16 (P < 0.05). No differences were found among the three groups in jugular LH levels during Days 1-18. Ovarian and uterine vein progesterone and oestradiol concentrations suggested that the elevated peripheral plasma concentrations of these steroids in pregnant animals were due mainly to increased ovarian production, but the embryo itself may have contributed to the oestradiol levels found at Day 17. Luteal tissues from pregnant animals contained more progesterone when incubated alone or with LH than tissues from cyclic animals (P < 0.06). However, LH-stimulated synthesis (incubated with LH minus incubated alone) was nearly identical for the three groups of tissues. Therefore, loss of responsiveness to LH does not appear to be the initial event in luteolysis. Concentrations of PGF in ovarian arterial plasma in cyclic and pregnant animals were similar at Day 18, although uterine vein concentrations were higher (P < 0.05) in the cyclic animals. Free arachidonic acid concentrations in corpora lutea at Day 18 were higher (P < 0.05) in cyclic than in pregnant animals. The data suggest that the embryo may produce one or more luteotrophic substances that stimulate increased progesterone secretion by the corpus luteum beginning as early as Day 10 of pregnancy.

Animals↗

Cyclic adenosine monophosphate signaling in the primate corpus luteum: maintenance of protein kinase A activity throughout the luteal phase of the menstrual cycle.

Recent studies from our laboratory (Endocrinology 136:4762-4768, 1995) demonstrating that the expression of cAMP-dependent nuclear transcription factor CREB (cAMP response element binding protein) is lost following ovulation in macaques has revealed a novel mechanism by which the cytoplasmic and nuclear actions the cAMP-protein kinase A (PKA) intracellular signaling system may be regulated independently. Implicit in this hypothesis is the assumption that PKA activity is maintained throughout the luteal phase of the menstrual cycle, yet to date there have been no published reports regarding PKA activity in the primate corpus luteum. PKA activity was assessed by the incorporation of 32P from radiolabeled ATP into a PKA-specific peptide substrate (kemptide) in the presence or absence of cAMP. Luteal cytosolic fractions were obtained from corpora lutea collected during the spontaneous luteal phase (days 3-5, 7-8, 10-11, 13-15, and postmenses) or obtained from animals on days 11 or 16 of the luteal phase after the animals received seven days of exogenous human CG (hCG) treatment. Examination of PKA activity in luteal slices from various aged CL maintained in short-term organ culture in the presence or absence of recombinant cynomolgus monkey LH was also performed. There were no significant differences in basal or cAMP-stimulated PKA activities in corpora lutea collected throughout the spontaneous luteal phase. Further, Western immunoblot analyses of the catalytic subunit of PKA (PKA C alpha) in corpora lutea collected throughout the luteal phase revealed immunoreactive protein bands with similar intensities. In vitro addition of recombinant cynomolgus LH and dibutyryl cAMP stimulated PKA activity in corpora lutea collected during the early, mid, and late luteal phases. In corpora lutea obtained from animals treated with hCG during the midluteal phase, basal PKA activity was decreased 65% as compared with untreated day 11 controls and in late luteal phase, hCG-exposed CL basal PKA activity was decreased 30% as compared with untreated day 16 controls. However, there were no measurable differences in cAMP-stimulated PKA activity in CL exposed to prior hCG treatment in vivo and Western immunoblot analyses for PKA C alpha in these tissues revealed immunoreactive protein bands that were comparable with corpora lutea collected from untreated animals. Further, immunoblot analyses for CREB in corpora lutea collected from hCG-treated animals revealed that CREB immunoreactivity remained undetectable following a treatment regimen with hCG that mimics early pregnancy. These results demonstrate that, although CREB expression ceases following ovulation, PKA activity is maintained throughout the luteal phase, which provides a mechanism by which the acute steroidogenic actions of LH may be separated from longer term trophic actions that may rely the transcriptional activity of CREB.

Aging↗

Roles of pulsatile release of LH in the development and maintenance of corpus luteum function in the goat.

The roles of the pulsatile release of LH in the functional development and maintenance of the corpus luteum (CL) during the estrus cycle in the goat were examined using a potent GnRH antagonist. In Experiment 1, to assess the inhibitory effects of the GnRH antagonist on the release of LH during the estrus cycle, 9 goats were divided into 3 groups. Goats in Group I received only saline on Days 0 (day of ovulation), 5, 10 and 15. Goats in Group II received the GnRH antagonist (50 microg/kg, s.c.) on the days mentioned for Group I to inhibit endogenous LH during the periods of luteal development and maintenance. Goats in Group III received saline on Days 0 and 5 and then the GnRH antagonist on Days 10 and 15 to inhibit LH during the period of luteal maintenance. Serial blood sampling took place on Days 1, 3, 5, 8, 13 and 18 to characterize the LH pulses. The LH pulses were observed throughout the estrus cycle in Group I but were completely abolished in Group II. In Group III, the pulsatile release of LH was observed from Day 1 to 8, but the LH pulses were completely abolished on Days 13 and 18. In Experiment 2, 16 goats were divided into the same 3 groups as in Experiment 1 to examine the effects of the GnRH antagonist on the luteal function. The concentration of progesterone in the plasma in Group I increased after ovulation, reached a maximum level around Day 12, and subsequently returned to the basal level on Day 17. The concentrations of progesterone in Group II rose after ovulation, but reached a plateau around Day 6 and maintained the level up to Day 9, then rapidly decreased from Day 9 to 10 to the basal level. The concentrations of progesterone in Group II were lower on Days 7 to 15 than those in Group I (P<0.01). The concentrations of progesterone in Group III increased after ovulation, reached a maximum level around Day 8, then dropped from Day 10 to 13 to the basal level. The concentrations of progesterone in Group III on Days 11 to 15 were lower than those in Group I (P<0.05 on Day 11, P<0.01 on Days 12 to 15). These results demonstrate that endogenous LH is essential for normal development and maintenance of the CL function during the estrus cycle in the goat. Further, this study suggests that while the functional maintenance of the caprine CL depends entirely on LH support, such functional dependence during early CL development is only partial.

Animals↗

Maintenance of the corpus luteum after uterine transfer of trophoblastic vesicles to cyclic cows and ewes.

One or two trophoblastic vesicles (0.4-2 mm diam.) from cow (Day 14) or ewe (Day 11-13) embryos without their disc were transferred, after culture for 24 h, into recipients. Each vesicle was transferred into the uterine horn ipsilateral to the CL by the cervical route in heifers and surgically in ewes on Day 12 of the oestrous cycle. In cows, daily measurements of plasma progesterone concentrations and checks for return to oestrus showed that the CL was maintained in 8 out of 12 recipients. These 8 cows had 25- to 37-day cycles while 4 recipient heifers returned to oestrus normally. Three recipients with an extended cycle were slaughtered. The dissected uterus showed that trophoblastic vesicles had developed in the uterine horns. In ewes, the serum progesterone curve, determined in each recipient, showed that the CL was maintained in 7 out of 12 recipients. These 7 ewes had 20- to 54-day cycles and the other 5 ewes had a normal cycle of 15-19 days comparable to that of 17.0 +/- 0.5 days for the 6 control ewes. Whenever the CL was maintained, high blood progesterone levels were followed by rapid luteolysis. In cattle and sheep, therefore, a trophoblastic vesicle transferred into the uterus can develop in vivo, secreting the embryonic signals when there is no embryonic disc control and transforming the cyclic CL into a CL of pregnancy in about 60% of the cases.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Relative roles of oestradiol and of the uterus in the maintenance of the corpus luteum in the pseudopregnant brown hare (Lepus europaeus).

Brown hares were made pseudopregnant by sterile matings or PMSG-hCG treatment (day of mating or hCG injection = Day 0 of pseudopregnancy). Progesterone secretion by the CL began 3-4 days after the ovulatory stimuli, reached maximum on Days 8 to 11 and decreased thereafter to reach low levels from Day 9 to 18, depending on the female. Cauterization of all large ovarian follicles on Day 7 resulted in an immediate luteolysis in young females, but had no effect in older ones. Oestradiol capsules implanted from Day 7 to Day 46 were able to maintain progesterone secretion until at least Day 30, in intact females as well as in females with all large follicles cauterized. Hysterectomy on Day 7 or 8 was followed by an immediate drop in progesterone concentrations; oestradiol capsules implanted at the time of hysterectomy prevented the drop in progesterone values, which remained elevated until Day 38. The induction of ovulation in females hysterectomized 2 months before resulted in CL of slightly shortened life-span. The injection of PGF-2 alpha on Day 7 of pseudopregnancy was followed by an immediate luteolysis. These results suggest that oestradiol secreted by the large ovarian follicles is the main luteotrophic factor in the brown hare. In old hares, the large amount of interstitial tissue could secrete oestrogens, and thus maintain pseudopregnancy. On Day 7 of pseudopregnancy, the uterus secretes a luteotrophic substance acting either directly on the ovary, or via the pituitary, to maintain oestradiol secretion by the follicles. In long-term hysterectomized females, the CL would be able to develop independently of any trophic substance, but for a reduced duration.

Animals↗

Maintenance of the corpus luteum of early pregnancy in the ewe. III. Differences between pregnant and nonpregnant ewes in luteal responsiveness to prostaglandin F2 alpha.

The effects of pregnancy and number of corpora lutea on luteal regression induced with prostaglandin F2 alpha (PGF2 alpha) were examined in 93 ewes. Bred and nonpregnant ewes were assigned randomly to receive a single im injection of PGF2 alpha: 0, 2, 4, 6, 8 or 10 mg/58 kg body weight. Injections were given on d 13 postestrus. The concentration of progesterone in serum 24 h after PGF2 alpha injection was affected by dose (P less than .001). The effect of pregnancy and the interaction of pregnancy with number of corpora lutea on levels of progesterone in serum were significant (P less than .05); therefore, data were partitioned according to pregnancy status and analyzed separately. There was an effect of number of corpora lutea on serum concentration of progesterone in pregnant (P less than .01) but not nonpregnant ewes (P greater than .10). Similar relationships among groups were observed for the concentration of progesterone in luteal tissue. In nonpregnant ewes the minimum dose of PGF2 alpha to produce a significant suppression of progesterone in serum (P less than .05) was 4 mg/58 kg body weight. In pregnant ewes with one or two corpora lutea, the minimum effective doses were 6 and 10 mg/58 kg body weight, respectively. The concentration of 13,14-dihydro-15-keto-prostaglandin F2 alpha (PGFM) in serum was related to the dose of PGF2 alpha injected. There were no differences in the concentration of PGFM in serum between pregnant and nonpregnant ewes either before or after injection. Corpora lutea of early pregnancy appear to be resistant to the luteolytic effect of PGF2 alpha.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Prolactin in maintenance of the corpus luteum of early pseudopregnancy in the golden hamster.

The role of prolactin in the maintenance of the corpus luteum of pseudopregnancy was studied in the golden hamster. Nine groups of seven to fourteen animals each received 1 mg bromocriptine at 11.00 h on days 1, 2 or 3 of pseudopregnancy (three groups for each day). On each day of treatment with bromocriptine, one group of hamsters was injected with bovine prolactin 4 h before bromocriptine, and one group received prolactin 4 h before bromocriptine for three consecutive days following treatment with bromocriptine. One group received bromocriptine only. These nine groups were compared with a control group of animals given 0.85% saline instead of bromocriptine and prolactin. Peripheral blood samples were taken from all hamsters at 11.00 h on days 3, 4, 5 and 6 of pseudopregnancy and plasma levels of progesterone were determined by radioimmunoassay. Luteolysis, indicated by a decline in progesterone level by 24 or 48 h after treatment with bromocriptine, occurred in all hamsters given bromocriptine alone, whether it was administered on day 1, 2 or 3. Pretreatment with a single dose of prolactin did not mitigate the bromocriptine-induced fall in progesterone. In the majority of cases, pretreatment with prolactin plus daily doses of prolactin maintained the progesterone at levels not different from saline-treated hamsters. These data suggest that prolactin is a necessary luteotrophin during early pseudopregnancy without which luteolysis ensues.

Animals↗

Endometrial responsiveness to oxytocin during diestrus and early pregnancy in pigs is not controlled solely by changes in oxytocin receptor population density.

These studies were performed to test the hypotheses that: 1) endometrial responsiveness to oxytocin (OT) in pig endometrium is associated with changes in OT receptor (OTr) population density resulting from corresponding regulation of OTr gene transcription, 2) endometrial responsiveness to OT is controlled solely through a mechanism involving changes in OTr population density, and 3) OTr population density and endometrial responsiveness to OT differ between diestrus and early pregnancy in pigs. In experiment 1, OTr population density and dissociation constant (Kd) in cyclic pigs were constant on Days 10-16 but increased (p < 0.05) between Days 10 and 12 of pregnancy before decreasing (p < 0.05) through Day 16. OT induced phosphoinositide (PI) hydrolysis and prostaglandin (PG) F2 alpha secretion in cyclic pigs only on Day 16 (p < 0.05), and during pregnancy only on Day 12 (p < 0.05). Activation of G protein by aluminum fluoride (AIF4-) treatment maximally stimulated (p < 0.05) PI hydrolysis and PGF2 alpha secretion in cyclic pigs on all days, indicating that downstream from the OTr, the PGF2 alpha secretory pathway was fully functional. During pregnancy, PI hydrolysis and PGF2 alpha secretion in response to AIF4- decreased (p < 0.01) on Days 14 compared to Days 10 and 12, and AIF4- did not stimulate PGF2 alpha release on Day 16. In experiment 2, abundance of OTr mRNA in cyclic pigs decreased between Days 0 and 5 before increasing between Days 5 and 12 (p < 0.05), but it was higher (p < 0.05) on Days 10-15 of pregnancy than on equivalent days in cyclic gilts. These results indicate that control of PGF2 alpha secretion in cyclic pigs appeared to occur primarily at the level of OTr coupling to G protein because changes in OTr number were not associated with increased sensitivity to OT or G-protein activation by AIF4-. During pregnancy, control was exerted at multiple levels, which included the OTr, G protein, phospholipase C, and subsequent aspects of the secretory pathway. The present study also indicated that endometrium was responsive to OT during luteolysis in cyclic pigs but not during corpus luteum maintenance in pregnant pigs.

Aluminum Compounds↗

Embryo-uterine interactions in pigs during week 2 of pregnancy.

The second week of pregnancy is a particularly critical period for embryonic survival in pigs. Within that time, conceptus oestrogen synthesis is initiated, spacing and final placement of conceptuses is completed, and the signal for extending the functional lifespan of the corpora lutea is received by the mother. There is also a marked increase in blood flow to the uterus and the uterine endometrium produces and secretes nutrient histotrophe. Conceptus-derived oestrogen has been implicated in many of these events. It is also during this period that the trophoblast elongates dramatically and the inner cell mass starts to differentiate into the embryo proper. Here, we critically review the evidence that oestrogen is the sole factor initiating long-term corpus luteum maintenance in pigs. We also review the functions and general properties of the major secretory proteins in histotrophe and the role of oestrogen in controlling their expression. It is now generally accepted that asynchrony within a litter underlies much of the losses of conceptuses that are otherwise genetically normal, but which are lagging in their development; however, the underlying mechanisms remain unclear. Here we hypothesize that oestrogenic compounds derived from more advanced conceptuses or provided prematurely, either by injection or in the diet, trigger a massive increase in uterine expression and secretion of retinol-binding protein laden with retinol. We propose that less developed, smaller conceptuses are least able to contend with the sudden exposure to this potential teratogen at a time when they are particularly susceptible to imbalance in retinol supply. Hence, even though their growth proceeds for a few days, their developmental potential is irrevocably compromised.

Animals↗

[The control of the development of the corpus luteum and its maintenance in cattle].

During its limited lifespan the bovine corpus luteum secretes large amounts of progesterone. The function of lipoproteins in maintaining corpus luteum activity and the role of high density lipoprotein (HDL) in development of the corpus luteum have been examined using luteal and granulosa cell cultures. Results show that luteal cells require lipoproteins for steroidogenesis, low density lipoprotein (LDL) being 3.5 times more potent than HDL. It is likely, however, that HDL is of more importance in vivo since concentrations in the blood are considerably higher than those of LDL. The results also show that luteal cells may be maintained in culture in the presence of HDL. Granulosa cells in culture show an increasing basal progesterone production. In addition they respond to HDL and dibutyryl cyclic AMP with a marked increase in progesterone production. This data suggests that granulosa cells in pre-ovulatory follicles are prevented from responding to HDL, perhaps through lack of receptors. Luteinization must allow granulosa cells to respond to HDL and, in addition, it provides an increase in availability of both HDL and LDL through the marked angiogenesis occurring at this time.

Animals↗

Evaluation of matrix metalloproteinases-1 and -3 concentrations in the tunica albuginea, the apical wall of atretic follicles and the corpus luteum of normal human ovaries.

Matrix metalloproteinases-1 (MMP-1) and -3 (MMP-3) are proteolytic enzymes involved in remodeling the ovarian extracellular matrix throughout the menstrual cycle. The aim of the present study was to evaluate the tissue concentrations of MMP-1 and MMP-3 in the apical wall of atretic follicles (androstenedione/estradiol ratio > 4), tunica albuginea dissected from the ovarian surface overlying areas devoid of follicles, corpus luteum, and tunica albuginea covering the corpus luteum. After extraction of MMPs from the tissue samples, their concentrations in the extracts were measured by enzyme-linked immunosorbent assays (ELISA). Significantly less MMP-1 was detected in the apical wall of atretic follicles compared to tunica albuginea taken from sites devoid of follicles. These data indicate that atresia is associated with relatively low concentrations of MMP-1 in the apical wall of the follicle. Moreover, there was a negative correlation between the amount of MMP-3 and the diameter of follicle. These data suggest that both MMPs play an important role in the final step of atresia. The amount of MMP-1 in the corpus luteum was several times lower than in the other tissues. This is likely due to stabilization of the extracellular matrix during the period of the corpus luteum maintenance. The concentration of MMP-3 did not differ significantly among the examined tissues.

Corpus Luteum↗

[Early diagnosis of pregnancy in the cow as assessed by milk ejection induced by luteal oxytocin].

The diagnosis of pregnancy described in this article is based on the observation of milk ejection which, in the case of corpus luteum maintenance, results from the release of luteal oxytocin induced by intravenous administration of a non luteolytic dose of PGF2 alpha. The tests were performed in 410 lactating cows, 18-22 days after insemination (mean 19.95 days, SD 0.75), 3 h prior to the evening milking. A cannula was placed in the left fore-teat; when the cisternal milk flow ceased, a small dose of a prostaglandin F2 alpha analogue (256 micrograms Dinolytic) was injected. If a corpus luteum was present, the alveolar milk flow (1,276 +/- 570 ml) started 86 +/- 35 s later and pregnancy could be presumed. If the corpus luteum was no longer functional, the milk flow did not start again and the cow was considered not pregnant. The accuracy of positive results, ie 72.3%, significantly exceeded that obtained by radioimmunoassay of plasma progesterone used as a reference method, ie 68.6% (P less than 0.05). By contrast, the accuracy of negative results was lower, ie 93.5 vs 100%, (P less than 0.05): of the 195 pregnant cows which proceeded to term, 10 did not respond to the milk ejection test. In spite of these reserves, the test has the advantage of being inexpensive, rapid (about 5 min) and easy to interpret, ie either the milk flows or it does not. In addition, it may be used early, because from day 20 after Al the reliability of prediction of non pregnant cows (67.0%) was significantly higher when compared to the use of progesterone radioimmunoassay (58.6%) (P less than 0.05). With such a test, non pregnant cows can be reinseminated immediately and not only after a 3-week period. This may contribute to reducing the length of the infertile period.

Animals↗