Circulating progesterone, progesterone-binding proteins and oestradiol-17 beta concentrations in the pregnant Cape porcupine, Hystrix africaeaustralis

Circulating concentrations of progesterone, progesterone-binding plasma proteins (PBPP) and oestradiol-17 beta in pregnant porcupines remained relatively low until Days 25-30 post coitum. Progesterone values peaked (102-180 ng/ml; N = 3) 42-60 days post coitum and the rapid increase in oestradiol-17 beta concentrations approximated that of progesterone with peak values (170-210 pg/ml) being attained 60-85 days post coitum. The pattern of PBPP synthesis, as suggested by circulating concentrations, was closely related to that of plasma progesterone, with values remaining low (less than 20 pmol/ml) until Day 31 post coitum, reaching peak levels at Days 50-56 and Days 73-77 post coitum. The production of PBPP during pregnancy is, as in related New World hystricomorph species, considered to be a mechanism which facilitates a reduction in the rate of progesterone metabolism during pregnancy.     

Fertility of sows injected with exogenous oestradiol and/or gonadotrophins to control post-weaning oestrus

In the first of two experiments 28 multiparous sows were allocated to one of the following treatments 2 days after weaning at approximately 35 days post partum: (1) untreated; (2) i.m. injection 10 μg oestradiol benzoate (OB)/kg body weight (b.wt.); and (3) i.m. injection 20 μg OB/kg b.wt. Sows were bred at first post-weaning oestrus and ovulation rate assessed at slaughter. The mean interval from weaning to oestrus in each group was: (1) 5.6 ± 0.2; (2) 4.7 ± 0.2; and (3) 4.7 ± 0.2 days; the mean ovulation rates in groups 1 and 2 (18.7 ± 0.6 and 17.4 ± 1.8, respectively) were significantly higher (P < 0.01) than that of 12.0 ± 1.7 for treatment 3 sows. Two untreated and one each of the treated sows were not cycling at slaughter.In the second experiment 75 multiparous sows weaned at 28 ± 3 days post partum (day 0) were evenly allocated with respect to parity to one of four treatment groups: (1) untreated; (2) i.m. injection 10 μg OB/kg b.wt. on day 2; (3) PG600 (400 iu PMSG + 200 iu hCG) injection subcutaneous day 0; and (4) combined PG600/OB treatment as in (2) and (3) above. Sows were bred naturally at the first post-weaning oestrus and fertility assessed at farrowing. Control animals had a significantly longer (P < 0.05) weaning to oestrus interval (4.53 ± 0.25 days) compared to treatment 2 (4.03 ± 0.13) treatment 3 (3.97 ± 0.12) and treatment 4 (3.81 ± 0.07) sows. Sows treated with PG600 alone showed a significant increase (P < 0.05) in numbers born live compared to pre-treatment values. A smaller and non-significant increase in numbers born live in control sows (probably related to increasing parity) was not observed in either OB- or PG600/OB-treated animals.These results suggest that with further modification of the treatments, a system may be developed for introducing fixed-time artificial insemination (AI) or mating as a means of controlling the reproductive performance of the weaned sow.     

Oestradiol and progesterone: soluble receptor levels and metabolism in the uterus of the ovariectomized ewe

Ovariectomized ewes received injections designed to mimic to some extent oestradiol and progesterone secretion during early pregnancy (maintenance progesterone), during oestrus (oestrous oestradiol) and during the luteal phase of the previous cycle (priming progesterone). The animals were killed at times equivalent to 1, 4 or 7 days after oestrus in those animals which had received oestrous oestradiol. The level of soluble oestradiol and progesterone receptors in whole uterus, and [3H]oestradiol and [3H]progesterone metabolism by uterus minces were measured. Oestradiol receptor level was highest on day 1 in those animals receiving oestrous oestradiol with no significant effect at any stage of the inclusion or omission of priming or maintenance progesterone. Progesterone receptor level was also high on day 1 in those animals receiving oestrous oestradiol with high levels maintained to day 4. Again, inclusion of priming or maintenance progesterone was without effect. In animals not receiving oestrous oestradiol the level of both receptors was uniformly low. Metabolism of [3H]oestradiol was low and not affected by treatment. [3H]Progesterone metabolism, although more variable, was also low and not affected by treatment.     

Fertility of the early postpartum, lactating domestic rabbit

Sixty-four crossbred primiparous lactating does each suckling six pups were allocated at random into four groups and were mated on either Day 1, 2, 3, or 4 post partum (where Day 0 = the day of parturition). They were subsequently killed on Day 10 post coitum (where Day 0 = the day of mating) to assess fertility. There were no significant differences between treatment groups in their mating response (97% overall), ovulation response (77% overall), implantation response (83% overall), implantation rate (8.7 overall), or preimplantation mortality rate (24% overall). Ovulation rate was significantly increased in does mated on Days 3 and 4 (13.3 and 13.1, respectively), compared with those mated on Day 1 (10.2, P<0.05) and Day 2 (9.6, P<0.01) post partum. From these results we conclude that fertility is high throughout the early postpartum period in the lactating rabbit.     

Postnatal development of the cholecystokinin-gastrin family of peptides in the brain and gut of rat

The average levels of CCK-8- and CCK-like peptides present in aqueous and acidic extracts of the brain of the newborn rat were, respectively, 5.3 and 2.9 pmol/g wet weight. These low levels increased 20- and 10-fold, respectively, so as to attain, at 25–30 days post partum, values comparable to those found in adult brain. There was also a rapid deposition of CCK-gastrin-like peptides in the whole gut between day 20 post coitum and day 10 post partum, when maximal concentrations were attained. Later, less CCK-gastrin peptides than other gut components were accumulated during the progressive weaning of the young rats, so that the average concentrations of CCK-gastrin-like peptides were, at day 30, similar to those observed in adult gut.     

Immunolocalization of the Vascular Endothelial Growth Factor Receptor-2 in the Subepicardial Mesenchyme of Hamster Embryos: IDentification of the Coronary Vessel Precursors

The earliest evIDence of the development of the cardiac vessels in mammals is the emergence of subepicardial blood islands, which are thought to originate from mesenchymal progenitors. In order to IDentify these progenitor cells, we have studied the immunohistochemical localization in the heart of Syrian hamster embryos of the type 2 vascular endothelial growth factor receptor, the earliest molecule known to be expressed in the vasculogenic cell lineage. Only a few immunoreactive subepicardial mesenchymal cells were present by 10 days post coitum. By 11 days post coitum, the subepicardial mesenchymal cells became abundant at the dorsal part of the ventricle, the atrioventricular and the conoventricular grooves. About 20% of cells were labelled with the antibody. Immunoreactive cells were isolated or formed pairs, short cords, rounded clusters or ring-like structures at the subepicardium or, occasionally, within the ventricular myocardium. Other labelled cells were simultaneously cytokeratin immunoreactive. By 12 days post coitum, most immunoreactive mesenchymal cells have been replaced by a capillary network. We propose that an active process of vascular differentiation occurs between 10 and 12 days post coitum in the subepicardium of this species, and it might be a suitable model for the study of vasculogenetic mechanisms.     

The effect of oestrogen and progesterone on maternal behaviour in rats.

Treatment or implantation of oestrone, oestradiol or progesterone on the third day postpartum into the ventromedial-arcuate region of the hypothalamus produced a complex pattern of changes in established maternal behaviour in rats. Progesterone implanted into the ventromedial-arcuate region of the hypothalamus increased the time spent nursing. In contrast, it was shortened by oestradiol given subcutaneously or implanted hypothalamically or extrahypothalamically, or by oestrone implanted into the hypothalamus. Oestradiol by an route used oestrone implanted into the hypothalamus caused marked reduction of pup weight and retrieval frequency. Progesterone had no such effect. Maternal care involving retrieval, sniffing, licking and nursing, expressed in seconds care, declined after hypothalamic and systemic oestradiol treatment. However, oestrone implanted into the hypothalamus increased seconds of care on days 9 and 10. Progesterone did not affect this function.     

Luteinizing hormone,total estrogens and progesterone secretion during lactation and after weaning in sows

Two experiments were conducted to determine changes in serum concentrations of LH, total free estrogens and progesterone before and after weaning in sows. Blood was collected either via indwelling anterior vena cava cannula or by venipuncture and serum hormones were measured by radioimmunoassay. In Exp. I, blood was collected at 15-min intervals for 4 hr on day 7 and day 21 postpartum from three sows on each day. In addition, individual samples were collected from 10 sows on days 4 and 14 postpartum and from 11 sows on days 1, 3 and 5 after weaning (day 23 postpartum). Serum LH ranged from .2 to .8 ng/ml during lactation and averaged 1.1 ± .7, 1.1 ± .7 and 2.7 ± .7 on days 1, 3 and 5 after weaning, respectively. Progesterone was low (< 1 ng/ml) during lactation and averaged 1.9 ± .3, .6 ± .3 and 1.2 ± .3 on days 1, 3 and 5 after weaning. Estrogens were variable during lactation, averaged 121 ± 36 pg/ml on day 1 after weaning and decreased thereafter. Estrus began on day 3 after weaning in 1 sow and on day 5 in the remaining 10 sows.In Exp. II, blood was collected from seven sows at 12 to 24 hr intervals from 2 days before until 5 days after weaning (day 26 postpartum). Mean serum LH was .7 ± .1 ng/ml during 48 hr before weaning and remained unchanged after weaning until day 3 when LH increased to 6.1 ± .8 ng/ml. Serum LH concentrations then declined to 1.3 ± .8 and .9 ± .8 ng/ml on days 4 and 5 after weaning. Total estrogens averaged 31 ± 4 pg/ml during 48 hr prior to weaning and 32 ± 4, 43 ± 17, 28 ± 1, 30 ± 2, 16 ± 2 and 18 ± 2 on days 0 to 5 after weaning. Progesterone increased from 1.0 ± .3 ng/ml 24 hr before weaning to 3.0 ± .3 at weaning and then remained low (< 1 ng/ml) until after ovulation when progesterone increased. Estrus began on day 4 after weaning in all seven sows.Results from these two experiments indicate that in sows: (1) LH is suppressed during early lactation (day 7), gradually increases during late lactation (day 21) and then reaches peak concentrations after weaning near the onset of estrus, (2) estrogens increase between weaning and estrus and decline thereafter, and (3) progesterone rises transiently at weaning and then increases after estrus and ovulation.     

Response of sows to oestradiol benzoate treatment after weaning at two stages of lactation

The timing and dosage of oestradiol benzoate injected after weaning was critical with respect to the pattern of behavioural oestrus and the ovarian stimulation achieved; treatment on the day of weaning (Day 0) and Day 1 with 60 micrograms oestradiol benzoate/kg body wt produced poor ovulatory responses and abnormal oestrous behaviour. Treatment on Day 2 with 30 micrograms oestradiol benzoate/kg resulted in consistent oestrus and ovulatory responses although the ovulation rates (10 . 6 +/- 1 . 1 in 3-week and 12 . 2 +/- 1 . 7 in 5-week weaned sows) were below those expected in fertile untreated sows weaned at these times. The timing of the preovulatory LH surge (53 . 6 +/- 2 h after oestradiol benzoate) was closely synchronized in all sows and a similar synchronous rise in plasma progesterone concentrations 100 +/- 4 h after oestradiol benzoate suggests a similar synchrony of ovulation. The magnitude of the LH and FSH responses to oestradiol benzoate were similar to those that occur in untreated sows and similar differences also existed in gonadotrophin secretion related to the length of lactation.     

Serum concentrations of oestradiol and progesterone during the normal oestrous cycle and early pregnancy in the lion (Panthera leo)

During a 6-month study period weekly serum samples demonstrated 9 oestradiol surges above 14 pg/ml (range 19-108 pg/ml) among 3 lionesses isolated from male lions. Intervals between peaks ranged from 3 to 8 weeks. Progesterone surges of more than 17 ng/ml (range 17-282 ng/ml) and lasting for 2-6 weeks were recorded after 7 of the oestradiol peaks. Sexual behaviour correlated well with the oestradiol peaks. Except for cornification following oestradiol peaks, there was no obvious vaginal cytology pattern at other times of the cycle. Pregnancy occurred after a 12-h contact with a male during behavioural oestrus. During gestation (108 days) oestradiol values remained low, while progesterone was elevated to 49 ng/ml within 12 h after mating, reaching a peak of 143 ng/ml at the 4th week, and remaining elevated during the next 2 months.     

Regulatory enzymes of carbohydrate and energy metabolism in the rabbit blastocyst.

The activities of phosphofructokinase, pyruvate kinase, citrate synthase and creatine kinase were determined in blastocysts from rabbits at 144 h post coitum and in similar blastocysts cultured for 24 h with or without oestradiol-17beta (1 microgrm/ml). There was a significant increase in all the enzymes during the 24-h culture period but oestradiol had no effect.     

Role of progesterone and oestradiol in the regulation of uterine oxytocin receptors in ewes.

The interaction between oestrogen and progesterone in the regulation of the uterine oxytocin receptor in sheep was evaluated by measuring the binding of oxytocin to membrane preparations of caruncular and intercaruncular endometrium and myometrium. Ovariectomized ewes were assigned in groups of five to each cell of a 4 x 2 factorial design. The four treatments were (a) vehicle (maize oil) for 12 days, (b) progesterone (10 mg day-1) for 9 days, (c) progesterone for 9 days followed by maize oil until day 12 and (d) progesterone for 12 days. The two oestradiol treatments consisted of the administration of implants in the presence or absence of oestradiol. The ewes were killed on day 10 (group b) or day 13 (groups a, c and d) for collection of uterine tissues. The response of the caruncular and intercaruncular endometrium to the treatments was similar. In the absence of oestradiol, treatment with progesterone continuously for either 9 or 12 days reduced the concentration of the oxytocin receptor in comparison with both the control and the progesterone withdrawal group (in which values were similar). The presence of oestradiol reduced the receptor concentrations in control and both 9- and 12-day continuous progesterone treatment groups, but enhanced the concentration in the progesterone withdrawal group. The myometrial oxytocin receptors responded in a similar way to those in the endometrium to progesterone treatment alone, but the addition of oestradiol produced no further effect. In conclusion, progesterone and oestradiol caused downregulation of the endometrial oxytocin receptor. On the other hand, progesterone withdrawal, similar to that which occurs during luteolysis, increased receptor density in the presence of oestradiol. Progesterone may influence the response of the myometrium to oxytocin by causing a reduction in receptor density.     

Transplacental Uptake of Glucose Is Decreased in Embryonic Lethal Connexin26-deficient Mice

Mice that harbor a targeted homozygous defect in the gene coding for the gap junctional protein connexin26 died in utero during the transient phase from early to midgestation. From day 10 post coitum onwards, development of homozygous embryos was retarded, which led to death around day 11 post coitum. Except for growth retardation, no gross morphological alterations were detected between homozygous connexin26-defective embryos and wild-type littermates.     

Effect of pregnancy, injection of oestradiol benzoate or hCG on steroid concentration and release by pig luteal cells

Corpora lutea were obtained from pig ovaries on Day 18 of pregnancy or pseudopregnancy. Pseudopregnancy was induced by the administration of oestradiol benzoate on Days 11-15 of the oestrous cycle or by the administration of hCG on Day 12. The luteal cells were prepared for morphometric analysis and investigation of steroid production in vitro by dispersion with 0.25% trypsin. A blood sample from each sow was collected at slaughter for measurement of progesterone, oestradiol-17 beta and testosterone. The concentrations of these steroids were also estimated in luteal tissue and in the medium after incubation. Progesterone concentration was significantly higher (P less than 0.01) in luteal tissue and in plasma of pregnant than of pseudopregnant sows. Testosterone content of luteal tissue from all sows was 20-fold higher than oestradiol, although plasma concentrations of these hormones were not different. The luteal cells from hCG-treated sows produced more progesterone (P less than 0.01) in vitro than did those from the other groups. The luteal cells from oestradiol-treated sows generally released smaller amounts of steroids during incubation. Treatment with hCG increased the proportion of large luteal cells and decreased the proportion of small luteal cells. These results demonstrate that hCG or oestradiol benzoate injections altered the steroidogenic activity of luteal cells and that treatment with hCG was also associated with changes in the diameter of the luteal cells and thus in the ratio of small to large luteal cells.     

Mouse Wnt receptor gene Fzd5 is essential for yolk sac and placental angiogenesis

Wnts are secreted signaling molecules implicated in various developmental processes and frizzled proteins are the receptors for these Wnt ligands. To investigate the physiological roles of frizzled proteins, we isolated and characterized a novel mouse frizzled gene Fzd5. Fzd5 mRNA was expressed in the yolk sac, eye and lung bud at 9.5 days post coitum. Fzd5 specifically synergized with Wnt2, Wnt5a and Wnt10b in ectopic axis induction assays in Xenopus embryos. Using homologous recombination in embryonic stem cells, we have generated Fzd5 knockout mice. While the heterozygotes were viable, fertile and appeared normal, the homozygous embryos died in utero around 10.75 days post coitum, owing to defects in yolk sac angiogenesis. At 10.25 days post coitum, prior to any morphological changes, endothelial cell proliferation was markedly reduced in homozygous mutant yolk sacs, as measured by BrdU labeling. By 10.75 days post coitum, large vitelline vessels were poorly developed, and the capillary plexus was disorganized. At this stage, vasculogenesis in the placenta was also defective, although that in the embryo proper was normal. Because Wnt5a and Wnt10b co-localized with Fzd5 in the developing yolk sac, these two Wnts are likely physiological ligands for the Fzd5-dependent signaling for endothelial growth in the yolk sac.     

Effects of reducing the remating interval after parturition on the fertility and plasma concentrations of luteinizing hormone, prolactin, oestradiol-17 beta and progesterone in lactating domestic rabbits

Primiparous crossbred does were remated on Day 1 (n = 15) or 14 (n = 25) post partum and killed on Day 10 post coitum to assess their fertility. Blood samples were taken during the pre- (0-12 h post coitum) and post- (1-10 days post coitum) ovulatory periods and plasma was assayed for luteinizing hormone (LH), prolactin, oestradiol-17 beta and progesterone. Ovulation response was significantly greater (P less than 0.01) and ovulation rate significantly lower (P less than 0.001) in does mated on Day 1 than in those mated on Day 14 post partum. Does failing to ovulate on Day 14 post partum exhibited no preovulatory LH surge and had significantly lower (P less than 0.05) premating concentrations of oestradiol-17 beta and prolactin than those ovulating at this time. No significant differences in hormone concentrations were observed during the preovulatory period between does ovulating on Days 1 and 14 post partum, with the exception of oestradiol-17 beta. Concentrations of this hormone were significantly lower (P less than 0.01) in does mated on Day 1, at 1 h post coitum. We conclude that (i) fertility was affected by the remating interval after parturition, (ii) ovulation failure was associated with an absence of the preovulatory LH surge and a reduction in premating concentrations of oestradiol-17 beta and prolactin and (iii) the lower ovulation rate in early lactation was apparently caused by a reduction in ovarian competence to respond to the gonadotrophic stimulus.     

Effects of weaning on concentrations of inhibin in follicular fluid and plasma of sows.

Changes in plasma and follicular fluid concentrations of inhibin were examined in sows after weaning at 28-32 days post partum. From 0 to 48 h after weaning, inhibin concentrations were 200-300 times higher in follicular fluid from small (less than 4 mm) and medium-large (greater than or equal to 4 mm) follicles than in ovarian venous plasma. Inhibin concentrations increased in follicular fluid from medium-large follicles at 24 and 48 h after weaning; concentrations in ovarian venous plasma were positively correlated with the number of medium-large follicles (r = 0.40) and with ovarian venous plasma concentrations of oestradiol (r = 0.61). Blood samples were collected for 30 days from sows (n = 6) that exhibited oestrus within 5 days after weaning and from sows (n = 5) that remained anoestrous for 11 days after weaning. Plasma inhibin concentrations rose in oestrous and anoestrous sows by 12 h and continued to rise for 60 h after weaning. Plasma inhibin concentrations rose further and were higher at 3.5-4.5 days after weaning in oestrous sows than in sows that remained anoestrous. After oestrus, plasma inhibin concentrations declined. At weaning, plasma concentrations of follicle-stimulating hormone (FSH) were higher in sows that subsequently exhibited oestrus than in sows that remained anoestrous. After weaning, plasma concentrations of FSH declined in both groups, reached a nadir at 2.5 days, and increased gradually in anoestrous sows; oestrous sows exhibited an FSH surge at oestrus. Plasma FSH returned to preweaning concentrations in both groups of sows at Days 7-8.(ABSTRACT TRUNCATED AT 250 WORDS)     

Embryo survival, progesterone profiles and metabolic responses to an increased feeding level during second gestation in sows

This study describes reproductive and metabolic responses in sows fed at two different feeding levels from day 3-35 of second gestation. After insemination, 37 sows were assigned to one of two treatments: 1) Control: 2.5 kg/day of a gestation diet; 2) Plus Feed 3.25 kg/day of a gestation diet (+30%). Sow weight, back fat and loin muscle depth were measured at farrowing, weaning, start of treatment, day 14 after start treatment and end of treatment. Frequent blood samples were taken for progesterone, luteinizing hormone (LH), glucose and insulin, insulin-like-growth-factor-1 (IGF-1), non-esterified-fatty-acids (NEFA) and urea analysis. At day 35 after insemination sows were euthanized and their reproductive tract collected to assess ovarian, embryonic and placental characteristics. Plus Feed sows gained 5.4 kg more weight and 0.9 mm more back fat and tended to be heavier at slaughter compared to Control sows (193 vs. 182 kg, P = 0.06). No difference in loin muscle gain was found. Treatment also did not affect vital embryonic survival, which was 72.1 ± 3.9% for Control and 73.4 ± 3.2% for Plus Feed sows, resulting in, respectively, 15.9 ± 0.9 and 15.7 ± 0.7 vital embryos. No effect of treatment on any of the ovarian, embryonic or placental characteristics was found. Progesterone profiles during the first month of gestation, and LH characteristics at day 14 of gestation were not different between treatments. Progesterone concentration was lower (P < 0.05) 3 h after feeding compared with the prefeeding level on days 7-11 after first progesterone rise for Plus Feed and on days 8-10 after first progesterone rise for Control sows. At day 15, preprandial glucose and insulin concentrations were not different between treatments, insulin peaked later (48 vs. 24 min) and at a higher concentration in Plus Feed than in Control sows. Furthermore, glucose area under the curve (AUC) tended to be lower (−171.7 ± 448.8 vs. 1257.1 ± 578.9 mg/6.2 h, P = 0.06, respectively) for Plus Feed vs. Control sows. IGF-1 concentration was not different between treatments, but NEFA concentrations were lower for Plus Feed vs. Control sows (149.5 ± 9.2 vs. 182.4 ± 11.9 μm/L, respectively, P = 0.04) and urea concentration tended to be higher in Plus Feed than in Control sows (4.3 ± 0.1 vs. 3.9 ± 0.1, respectively, P = 0.13). None of the metabolic parameteres were related to reproductive measures. In conclusion, feeding 30% more feed from day 3 till d 35 of second gestation increased weight gain and resulted in lower NEFA concentrations, but did not affect progesterone, LH or IGF-1 and embryonic and placental characteristics.     

Oestrone Sulphate Measurements for the Prediction of Small or Large Litters in Pigs

Serum from 88 pregnant sows and gilts was sampled 24 and 28 days after their first insemination or mating day. The oestrone sulphate (E1S) concentration in the samples was assessed with a commercially available radioimmunoassay kit modified for use with swine serum. The first aim was to test whether it was possible to predict litters of total number <10 piglets at term. The second aim was to compare the use of day 24 or day 28 samples, or of both, in this prediction.