Developmental regulation of placental insulin-like growth factor (IGF)-II and IGF-binding protein-1 and -2 messenger RNA expression during primate pregnancy.

The present study was conducted to determine the developmental expression of placental insulin-like growth factor (IGF)-II, IGF-binding protein (IGFBP)-1 and -2, and IGF-II receptor mRNA expression during baboon pregnancy and whether estrogen, the levels of which increase with advancing pregnancy, regulates placental trophoblast IGF-II mRNA expression. Levels of the IGF-II 6.1-kilobase (kb) and 4.9-kb mRNA transcripts determined by Northern blot analysis progressively increased three- to fourfold in placental syncytiotrophoblast and whole-villous tissue between early (Day 60), mid (Day 100), and late (Day 170) baboon gestation (term = 184 days). In contrast, syncytiotrophoblast IGFBP-1 and -2 mRNA levels decreased, and IGF-II receptor mRNA expression remained relatively constant, with advancing baboon pregnancy. Placental cytotrophoblast IGF-II mRNA levels determined by competitive reverse transcription-polymerase chain reaction on Day 54 of gestation were increased (P < 0.05) almost twofold at 18 h after acute administration of estradiol to baboons, whereas long-term estrogen treatment had no effect. We propose that these changes in trophoblast IGF expression would provide a mechanism for enhancing net bioavailability and bioreactivity of IGF-II locally to promote the growth and development of the placenta and, consequently, of the fetus during primate pregnancy.

Concentrations of progesterone and oxytocin receptors in endometrium of postpartum cows expected to have a short or normal oestrous cycle.

Luteal lifespan is short after first postpartum ovulation in early-weaned beef cows unless cows are pretreated with a progestogen. Regression of the short-lived corpus luteum in the postpartum beef cow is due to a premature release of prostaglandin F2 alpha (PGF2 alpha) from the uterus. The premature release of PGF2 alpha may be mediated through lower concentrations of receptors for progesterone, higher concentrations of oxytocin receptors, or both, in the endometrium. Thirty-one beef cows were randomly assigned to four groups at parturition. Calves from cows assigned to the short cycle group (n = 6; control) and the short cycle/endometrium group (n = 10) were weaned at 30-32 days post partum. Cows in the normal cycle group (n = 5; control) and the normal cycle/endometrium group (n = 10) received norgestomet implants for 9 days beginning 21-23 days post partum and calves were weaned at implant insertion. Duration of oestrous cycle (x +/- SEM; P < 0.01) following first postpartum ovulation for the short cycle group was 11.5 +/- 1.9 days compared with 18.8 +/- 0.6 days for the normal cycle group. On day 5 following first postpartum ovulation, cows in the short cycle/endometrium and the normal cycle/endometrium groups were hysterectomized and endometrial tissue collected for measurement of progesterone and oxytocin receptors. Mean number of total progesterone receptors per cell was lower (P < 0.05) in the short cycle/endometrium group than in the normal cycle/endometrium group. Mean concentration of oxytocin receptors (fmol mg-1 protein) in the short cycle/endometrium group was higher (P < 0.05) than that in the normal cycle/endometrium group.(ABSTRACT TRUNCATED AT 250 WORDS)

Basis of norgestomet action as a progestogen in cattle.

Norgestomet is the progestational component present in Syncro-Mate-B, which is used to synchronize estrus in cattle. In post-partum cows, luteal phases anticipated to be short following first ovulation are of normal length when cows are pretreated with norgestomet. Because Syncro-Mate-B is used experimentally as a progestogen to affect uterine function, these studies were conducted to investigate how norgestomet acts at the level of the uterus. Receptor binding assays and a sensitive estrogen bioassay in tissue culture were used to address the possibility that some effects of norgestomet might be mediated through interaction of this compound with steroid hormone receptors other than the progesterone receptor (rP). The source of receptors was high-speed cytosol, prepared from bovine uterine endometrium, which was obtained from cyclic cows. Results of single-point and complete competition analyses comparing norgestomet and progesterone indicated that norgestomet competed even more effectively than did progesterone for specific binding of [3H]progesterone to rP. Results of similar studies, which compared the abilities of norgestomet and diethylstilbestrol to compete with [3H]estradiol for binding by uterine endometrial estrogen receptors (rE), provided no evidence for norgestomet competitive binding to rE. In MCF-7 breast cancer cell bioassays, norgestomet showed weak estrogenic activity, but only at concentrations greater than 1 micromolar. Finally, norgestomet did not compete with [3H]triamcinolone acetonide when present in an 100-fold excess, and only competed with [3H]dexamethasone for binding by endometrial glucocorticoid receptors (rG) when present in the micromolar range. We conclude that, at the concentrations used in synchronizing estrus, norgestomet interacts with bovine endometrium as a progestogen and that its biological actions occur through binding of this compound to rP present in target tissues.

In vitro secretion of prostaglandins from endometrium of postpartum beef cows expected to have short or normal luteal phases.

The objective of this study was to characterize endometrial secretion (in vitro) of prostaglandin F (PGF), 15-keto-13,14-dihydro-prostaglandin F2 alpha (PGFM), prostaglandin E2 (PGE2), and 6-keto-prostaglandin F1 alpha (6-keto-PGF1 alpha) on Day 5 following the first postpartum estrus of cows anticipated to have a short compared to a normal estrous cycle. Twenty-seven beef cows were randomly assigned into four groups. The Short Cycle (n = 6; control) and Short Cycle/Explant (n = 8; endometrial explants) groups had their calves weaned at 30-32 days postpartum. The Normal Cycle (n = 5, control) and Normal Cycle/Explant (n = 8; endometrial explants) groups received norgestomet (progestin) implants for 9 days beginning 21-23 days postpartum, and calves were weaned at implant insertion. Estrous cycle length (mean +/- SE; p less than 0.01) for the Short Cycle group was 11.5 +/- 1.9 days compared to 18.8 +/- 0.6 days for the Normal Cycle group. On Day 5 following the first postpartum estrus, cows in the Short Cycle/Explant and Normal Cycle/Explant groups were hysterectomized, and endometrial explants were incubated in Earle's Balanced Salt solution/Medium 199 for 90 min with or without arachidonic acid (AA) in the presence of three levels of oxytocin. Mean concentrations of PGF and PGFM were combined to obtain a value for total PGF. Concentrations of total PGF, PGE2 (from explants without AA treatment), and 6-keto-PGF1 alpha in medium of the Short Cycle/Explant group were higher (p less than 0.01) than in medium of the Normal Cycle/Explant group.(ABSTRACT TRUNCATED AT 250 WORDS)

Oxytocin-induced release of prostaglandin F2 alpha in postpartum beef cows: comparison of short versus normal luteal phases.

The first postpartum ovulation after early weaning of calves (30 35 days of age) from cows is normally followed by a short luteal phase (6 10 days) unless the animals are pretreated with a progestogen (e.g. norgestomet). Reduced luteal lifespan in cattle is reportedly due to the premature release of a luteolysin (presumably prostaglandin F2 alpha [PGF2 alpha]). Therefore, the objective was to determine if oxytocin-induced release of PGF2 alpha (measured by the stable PGF2 alpha metabolite, 15-keto-13,14-dihydro PGF2 alpha [PGFM]) was greater for cows having a short compared to a normal luteal phase on Day 5 following the first postpartum estrus (Day 0). Thirty postpartum beef cows were randomly assigned into three groups (n = 10 per group) expected to have short (Short d 5) or normal (Norgestomet d 5 and Norgestomet d 16) luteal phases. Cows in Norgestomet d 5 and d 16 groups received Norgestomet (progestogen) implants for 9 days beginning 21 23 days postpartum. On Day 5 (Short d 5 and Norgestomet d 5) or Day 16 (Norgestomet d 16) following first postpartum estrus, each animal was injected (i.v.) with 100 IU oxytocin. In addition, cows in the Short d 5 group were subdivided into two groups following second estrus (normal luteal phase, n = 5 per group) to receive 100 IU oxytocin on Day 5 (Normal d 5) or 16 (Normal d 16), respectively. Estrous cycle length (means +/- SE) for cows in the Short d 5 group (8.7 +/- 0.4 days) was shorter (p less than 0.01) than for cows in all other groups (21.1 +/- 0.3 days).(ABSTRACT TRUNCATED AT 250 WORDS)

Ovarian inhibition of peripheral plasma concentration of follicle stimulating hormone in prepuberal Holstein heifers.

The objective of this study was to determine effects of age and castration on follicle stimulating hormone (FSH) secretion in prepuberal heifers. In experiment 1, twelve heifers were bilaterally ovariectomized at 3, 6, or 9 months of age (n = 4/group). Blood was collected at 10 min intervals for 8 hr at 1 week before ovariectomy and 1 and 4 weeks after ovariectomy. Frequency, amplitude and duration of FSH pulses were calculated. Mean plasma concentration of FSH (ng/ml), and frequency (pulses/8 hr), amplitude (ng/ml), and duration (min/pulse) of FSH pulses were not altered by age. Mean concentration of FSH increased (P less than .01) from 1 week before to 1 week and 4 weeks after ovariectomy, respectively, in all age groups. Pulse frequency increased (P less than .05) from 1 week before ovariectomy to 4 weeks after ovariectomy in 3 month old heifers, from 1 week before to 4 weeks after ovariectomy in 6 month old heifers, and from 1 week before to 1 week and 4 weeks after ovariectomy in 9 month old heifers. In experiment 2, twelve heifers were bilaterally ovariectomized at 3, 6 or 9 weeks of age (n = 4/group). Sample collection and measurement of mean concentration of FSH were the same as in experiment 1. Mean concentration of FSH increased (P less than .01) from 1 week before to 1 and 4 weeks after ovariectomy in heifers ovariectomized at 6 and 9 weeks of age.(ABSTRACT TRUNCATED AT 250 WORDS)