Production and secretion of progesterone in vitro and presence of platelet activating factor (PAF) in early pregnancy of the marsupial, Macropus eugenii

1993 ◽  
Vol 5 (1) ◽  
pp. 15 ◽  
Author(s):  
T Kojima ◽  
LA Hinds ◽  
WJ Muller ◽  
C O'Neill ◽  
CH Tyndale-Biscoe

The corpus luteum (CL) of the tammar is suppressed by lactation. Removal of the sucking pouch young induces reactivation of the CL which produces a peak in plasma progesterone concentration on Day 5, 6 or 7; reactivation of the embryo after diapause precedes by one day this transient peak of progesterone. This study examines progesterone production and secretion in vitro by the CL and the production of platelet activating factor (PAF) by the endometrium during this stage of pregnancy (Days 0 and 3-8). Blood samples were collected twice daily to determine for each animal (n = 28) at autopsy the relationship of PAF production to the day of the progesterone peak. Significant changes in basal plasma progesterone concentrations were used to define four groups according to the time the animals were killed: Group A, Day 0; Group B, other animals showing no significant change; Group C, peripheral progesterone concentration increasing; Group D, peripheral progesterone concentration decreasing after the peak. At autopsy on successive days, blood samples were taken from the heart, caudal vein and uterine branch of the ovarian vein, and CL and endometria were prepared for explant culture. Progesterone contents of pre- and post-incubation luteal tissues and of medium were determined at 4, 12 and 24 h to estimate production and secretion rates. The values for all progesterone parameters from animals in Group C were significantly higher than in the other groups. It is concluded that the progesterone peak results from a change in rate of production and secretion of progesterone by the CL, one day before the peak in peripheral plasma, and that this change could provide the first signal to the uterus. Endometrium was incubated for 24 h and the medium assayed for PAF. High concentrations of PAF were detected in half the animals in Groups B and C, that is, before or at the time progesterone was increasing. This is the first measurement of PAF in any marsupial and the first demonstration of the release of PAF by the endometrium. The detection of PAF provides a candidate for an endometrial signal that is responsive to the rising progesterone concentration in the utero-ovarian vein draining the CL-bearing ovary and that may stimulate reactivation of the blastocyst.

mBio ◽  
2020 ◽  
Vol 11 (6) ◽  
Author(s):  
Lindsey R. Burcham ◽  
Yoann Le Breton ◽  
Jana N. Radin ◽  
Brady L. Spencer ◽  
Liwen Deng ◽  
...  

ABSTRACT Nutritional immunity is an elegant host mechanism used to starve invading pathogens of necessary nutrient metals. Calprotectin, a metal-binding protein, is produced abundantly by neutrophils and is found in high concentrations within inflammatory sites during infection. Group B Streptococcus (GBS) colonizes the gastrointestinal and female reproductive tracts and is commonly associated with severe invasive infections in newborns such as pneumonia, sepsis, and meningitis. Although GBS infections induce robust neutrophil recruitment and inflammation, the dynamics of GBS and calprotectin interactions remain unknown. Here, we demonstrate that disease and colonizing isolate strains exhibit susceptibility to metal starvation by calprotectin. We constructed a mariner transposon (Krmit) mutant library in GBS and identified 258 genes that contribute to surviving calprotectin stress. Nearly 20% of all underrepresented mutants following treatment with calprotectin are predicted metal transporters, including known zinc systems. As calprotectin binds zinc with picomolar affinity, we investigated the contribution of GBS zinc uptake to overcoming calprotectin-imposed starvation. Quantitative reverse transcriptase PCR (qRT-PCR) revealed a significant upregulation of genes encoding zinc-binding proteins, adcA, adcAII, and lmb, following calprotectin exposure, while growth in calprotectin revealed a significant defect for a global zinc acquisition mutant (ΔadcAΔadcAIIΔlmb) compared to growth of the GBS wild-type (WT) strain. Furthermore, mice challenged with the ΔadcAΔadcAIIΔlmb mutant exhibited decreased mortality and significantly reduced bacterial burden in the brain compared to mice infected with WT GBS; this difference was abrogated in calprotectin knockout mice. Collectively, these data suggest that GBS zinc transport machinery is important for combatting zinc chelation by calprotectin and establishing invasive disease. IMPORTANCE Group B Streptococcus (GBS) asymptomatically colonizes the female reproductive tract but is a common causative agent of meningitis. GBS meningitis is characterized by extensive infiltration of neutrophils carrying high concentrations of calprotectin, a metal chelator. To persist within inflammatory sites and cause invasive disease, GBS must circumvent host starvation attempts. Here, we identified global requirements for GBS survival during calprotectin challenge, including known and putative systems involved in metal ion transport. We characterized the role of zinc import in tolerating calprotectin stress in vitro and in a mouse model of infection. We observed that a global zinc uptake mutant was less virulent than the parental GBS strain and found calprotectin knockout mice to be equally susceptible to infection by wild-type (WT) and mutant strains. These findings suggest that calprotectin production at the site of infection results in a zinc-limited environment and reveals the importance of GBS metal homeostasis to invasive disease.


2009 ◽  
Vol 21 (1) ◽  
pp. 101 ◽  
Author(s):  
O. A. Bogle ◽  
D. Ambati ◽  
R. P. Davis ◽  
G. P. Adams

The presence of an ovulation-inducing factor (OIF) in the seminal plasma of llamas and alpacas (reflex ovulators) and cattle (spontaneous ovulators) has been reported previously (Ratto MH et al. 2006 Theriogenology 66, 1102–1106). The presence of this protein in unrelated species supports the hypothesis that OIF is a conserved factor among species. The objectives of this study were to determine if OIF was present in equine and porcine seminal plasma, and whether the proportion of test animals (llamas) that ovulated in response to treatment with seminal plasma was related to dose. In Experiment 1, female llamas were assigned randomly to four groups (n = 8 or 9 per group) and treated intramuscularly with 1 mL llama seminal plasma (positive control), 3 mL equine seminal plasma, 3 mL porcine seminal plasma, or 2 mL saline (negative control). Ovulation and maximum corpus luteum diameter were compared using ultrasonography and confirmed with blood samples taken on Day 7 (Day 0 = day of treatment) to determine plasma progesterone concentration. The diameter of the preovulatory follicle at the time of treatment did not differ among groups. Equine seminal plasma induced ovulations in 3/8 (38%) llamas compared to 0/8 (0%) llamas treated with saline or porcine seminal plasma (P = 0.1). The proportion of females that ovulated was lower in the equine group (P < 0.01) compared with those animals treated with llama seminal plasma (9/9; 100%). Of the animals that ovulated, maximum CL diameter did not differ between llama and equine seminal plasma-treated groups (mean ± SEM; 11.1 ± 1.1, 11.5 ± 1.5, respectively). Similarly, progesterone concentrations were not different among llamas treated with llama seminal plasma or equine seminal plasma (mean ± SEM; 3.1 ± 0.4, 3.7 ± 1.2, respectively). The design of Experiment 2 was the same, but the dose of equine and porcine seminal plasma was increased to 8 mL and 10 mL, respectively. The proportion of females that ovulated was less (P < 0.05) in equine (2/9) and porcine (3/9) seminal plasma groups compared with the group treated with llama seminal plasma (9/9). There were no ovulations detected in llamas treated with saline (0/8). Although differences between equine, porcine, and negative control groups did not reach significance, results provide some evidence for the presence of OIF in equine and porcine seminal plasma. The effect of dose of equine and porcine seminal plasma is equivocal, suggesting that the concentration of OIF in the seminal plasma of these species may be very low and the optimal dose for inducing ovulation in test animals had not been reached. Research supported by the Natural Sciences and Engineering Council of Canada.


1984 ◽  
Vol 38 (1) ◽  
pp. 83-89 ◽  
Author(s):  
E. A. Amoah ◽  
M. J. Bryant

ABSTRACTThirty-six British Saanen female kids, born between either 20 March and 6 April (E) or 10 April and 16 June (L), were reared under three different patterns of lighting — a constant 18-h light: 6-h dark (C); or a weekly step-wise reduction from 18-h to 10-h light: 14-h dark over 10 weeks, commencing at either 6 (R6) or 10 (R10) weeks of age. Vasectomized males were used to test for oestrus; blood samples were collected to determine plasma progesterone concentration. Thirty-three goats attained puberty; all but two kids showed oestrus at first ovulation. The mean ( ± s.d.) age and live weight at puberty were 174·6 (± 4·1) and 141·7 (± 4·3) days (P < 0·001), and 28·6 (± 0·9) and 22·8 (± 1·0) kg (P < 0·001), for E and L treatments, respectively. For R6, R10 and C treatments, the ages and weights wer e 151·6 (± 4·7), 160·4 (± 5·2) and 164·7 (± 4·9) days (NS), respectively, and 24·5 (± 1·1), 25·3 (± 1·2) and 27·8 (± 1·1) kg (NS), respectively. A significant date of birth × lighting-pattern treatment interaction was found for date at puberty (P < 0·001), with puberty occurring earlier for E than L kids only when a reduction in daylength occurred. Puberty occurred earlier on R6 than R10 treatments only for L kids.


1977 ◽  
Vol 75 (1) ◽  
pp. 127-136 ◽  
Author(s):  
J. S. WALTON ◽  
JUDITH R. McNEILLY ◽  
A. S. McNEILLY ◽  
F. J. CUNNINGHAM

The plasma concentrations of FSH, LH, prolactin and progesterone were measured throughout anoestrus and during the resumption of cyclic activity in two groups of ewes. Group A was maintained under conditions of natural daylength throughout the experiment, whereas Group B was exposed to 6 h of light and 18 h of darkness, the change being made abruptly on the longest day. In those ewes kept on short days, oestrus and ovulation occurred 18·3 days (P < 0·05) and 23·4 days (P < 0·001) earlier than in the ewes under conditions of natural daylength. Ovulation preceded oestrus by 18·6 days and 23·1 days in Groups A and B respectively. The occurrence of ovulation was detected by the determination of plasma progesterone concentrations. In all ewes, progesterone levels were basal until the first ovulation when the pattern of secretion was typical of that seen during the oestrous cycle. In the 4 days before ovulation, the plasma progesterone concentration increased slowly to reach a maximum of 0·66 ± 0·12 (s.e.m.) ng/ml on day − 1. The first ovulation was associated with a substantial surge of LH. Similar release of LH, thought to be related to the increased progesterone secretion, was also observed on day −5. Sporadic release of LH was also found before this time in some animals. Plasma concentrations of FSH fluctuated randomly throughout anoestrus and during the transition to established oestrous cycles. These changes were not apparently related to ovulation. Throughout anoestrus, prolactin concentrations were raised but always declined before the time of ovulation. The fall in prolactin concentrations occurred sooner in the ewes on short days, pointing to a relationship with the decreased daylength. These data suggest that the return to oestrous cycles may be brought about by the removal of an antigonadotrophic effect exerted by high concentrations of prolactin in the blood during anoestrus.


1969 ◽  
Vol 45 (3) ◽  
pp. 449-457 ◽  
Author(s):  
J. M. BASSETT ◽  
TANA J. OXBORROW ◽  
I. D. SMITH ◽  
G. D. THORBURN

SUMMARY The progesterone concentration in the peripheral plasma of ewes throughout pregnancy has been determined by a protein-binding method. Plasma progesterone concentrations during the first 50 days of pregnancy (2–3 ng./ml.) were not significantly higher than peak concentrations during the luteal phase in cycling non-pregnant ewes, but there was no decrease in the concentration 15–20 days after mating as occurs in non-pregnant ewes. Between 50 and 120 days after mating the plasma progesterone concentration increased steadily to values 2–5 times that found in early pregnancy. These high concentrations were maintained until lambing. A decrease in progesterone concentration during the week preceding lambing was usually, but not always, observed. Mean plasma progesterone concentrations during the last 50 days of pregnancy in ewes with twins were approximately twice those in ewes with a single foetus.


Rangifer ◽  
1991 ◽  
Vol 11 (2) ◽  
pp. 13 ◽  
Author(s):  
Erik Ropstad ◽  
Dag Lenvik

A total of 126 reindeer of about 7 months of age, were isolated from a flock at the end of the breeding season. The animals were treated either with 12.5 mg prostaglandin F2alpha (n = 41) or 0.25 mg cloprostenol (n = 50). Thirty-five animals were left untreated. Blood samples were collected before treatment and 2 Vi days later and the plasma progesterone concentrations were determined. A significant fall in progesterone concentration was seen in both treatment groups. A large proportion of animals responded to treatment with cloprostenol than with prostaglandin F2alpha. It was concluded that prostaglandins can be used to induce luteolysis in reindeer.


1974 ◽  
Vol 27 (6) ◽  
pp. 659 ◽  
Author(s):  
AR Gleeson ◽  
GD Thorburn

A competitive protein-binding technique was used to measure progesterone concentrations in the peripheral plasma of pregnant ewes. Neither haemolysis of blood nor thawing of plasma samples affected plasma progesterone concentration. Blood samples should be chilled immediately upon collection but subsequent to centrifugation immediate chilling of the plasma samples is not critical. No consistent diurnal variation in progesterone concentrations was evident but there was large apparently random day-to-day variation in progesterone concentrations for any ewe. Although a significant positive correlation was found between endogenous progesterone and corticosteroid concentrations, the present study failed to correlate experimentally elevated plasma corticosteroid concentrations with progesterone concentrations. Progesterone concentrations varied greatly between ewes at the same stage of pregnancy.


1973 ◽  
Vol 58 (2) ◽  
pp. 219-225 ◽  
Author(s):  
K. P. McNATTY ◽  
K. J. A. REVFEIM ◽  
A. YOUNG

SUMMARY Progesterone concentrations in peripheral plasma were measured once daily during one oestrous cycle in each of eight sheep. In addition, on days 4–5, 8–9, 12–13 and 15–16 of the oestrous cycle, blood samples were collected at 30-min intervals throughout each 24-h period. A total of three ewes was sampled in each 24-h period and the same three animals were not bled again for at least 1 week. Plasma progesterone concentrations in all the ewes fluctuated considerably throughout each 24-h period. The within-sheep within-day variations observed in peripheral progesterone concentrations were compared with the between-sheep within-day variations and the within-sheep between-day variations previously reported. It is concluded that these previously reported variations in peripheral plasma progesterone concentration could be attributed to within-day variations in each animal. On days 8–9 and 12–13 of the oestrous cycle there were significantly higher concentrations of progesterone in plasma during the hours of daylight than during the hours of darkness. In contrast, progesterone concentrations on days 4–5 and 15–16 were not found to be significantly different between the hours of daylight and darkness. These results suggest that diurnal changes in peripheral plasma progesterone concentration occur during the luteal phase of the ovine oestrous cycle.


1971 ◽  
Vol 66 (3) ◽  
pp. 471-477 ◽  
Author(s):  
Anne Kristine Blom ◽  
Olav Lyngset

ABSTRACT Plasma progesterone concentration at different stages of pregnancy was measured in the blood obtained from the uterine artery (6 goats), the uterine vein (6 goats), and the ovarian vein (11 goats). Progesterone concentration was also measured in the peripheral venous blood before and after extirpation of the ovaries and the uterus (16 goats). The concentration of progesterone was found to rise gradually in the peripheral and ovarian vein blood to reach a maximum at 90 days of pregnancy. This was followed by a gradual decline with consistent low values of about 7 ng prog./ml 3–4 days before parturition. A significant decrease in peripheral progesterone concentration was found 10 minutes after removing the ovaries and the uterus. In two goats sampled before and after parturition, the plasma progesterone concentration was found to decrease before parturition and to remain low for at least three days after parturition.


2020 ◽  
Author(s):  
Lindsey R. Burcham ◽  
Yoann Le Breton ◽  
Jana N. Radin ◽  
Brady L. Spencer ◽  
Liwen Deng ◽  
...  

AbstractNutritional immunity is an elegant host mechanism used to starve invading pathogens of necessary nutrient metals. Calprotectin, a metal binding protein, is produced abundantly by neutrophils and is found in high concentrations within inflammatory sites during infection. Group B Streptococcus (GBS) colonizes the gastrointestinal and female reproductive tracts and is commonly associated with severe invasive infections in newborns such as pneumonia, sepsis, and meningitis. Though GBS infections induce robust neutrophil recruitment and inflammation, the dynamics of GBS and calprotectin interactions remain unknown. Here we demonstrate that disease and colonizing isolate strains exhibit susceptibility to metal starvation by calprotectin. We constructed a mariner transposon (Krmit) mutant library in GBS and identified 258 genes that contribute to surviving calprotectin stress. Nearly 20% of all underrepresented mutants following treatment with calprotectin, are predicted metal transporters, including known zinc systems. As calprotectin binds zinc with picomolar affinity, we investigated the contribution of GBS zinc uptake to overcoming calprotectin-imposed starvation. Quantitative RT-PCR revealed a significant upregulation of genes encoding zinc-binding proteins, adcA, adcAII, and lmb, following calprotectin exposure, while growth in calprotectin revealed a significant defect for a global zinc acquisition mutant (ΔadcAΔadcAIIΔlmb) compared to the GBS WT strain. Further, mice challenged with the ΔadcAΔadcAIIΔlmb mutant exhibited decreased mortality and significantly reduced bacterial burden in the brain compared to mice infected with WT GBS; this difference was abrogated in calprotectin knockout mice. Collectively, these data suggest that GBS zinc transport machinery are important for combatting zinc-chelation by calprotectin and establishing invasive disease.ImportanceGBS asymptomatically colonizes the female reproductive tract but is a common causative agent of meningitis. GBS meningitis is characterized by extensive infiltration of neutrophils, carrying high concentrations of calprotectin, a metal chelator. To persist within inflammatory sites and cause invasive disease, GBS must circumvent host starvation attempts. Here, we identified global requirements for GBS survival during calprotectin challenge, including known and putative systems involved in metal ion transport. We characterized the role of zinc import in tolerating calprotectin stress in vitro, and in a mouse model of infection. We observed that a global zinc-uptake mutant was less virulent compared to the parental GBS strain and found calprotectin knockout mice to be equally susceptible to infection by WT and mutant strains. These findings suggest that calprotectin production at the site of infection results in a zinc-limited environment and reveals the importance of GBS metal homeostasis to invasive disease.


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