Regulation of oxytocin, oestradiol and progesterone receptor concentrations in different uterine regions by oestradiol, progesterone and oxytocin in ovariectomized ewes

1996 ◽  
Vol 151 (3) ◽  
pp. 375-393 ◽  
Author(s):  
D C Wathes ◽  
G E Mann ◽  
J H Payne ◽  
P R Riley ◽  
K R Stevenson ◽  
...  
Keyword(s):  
Epithelial Cells ◽  
Low Dose ◽  
Progesterone Receptors ◽  
Cell Types ◽  
Oxytocin Receptor ◽  
Luminal Epithelium ◽  
Time Points ◽  
Progesterone Treatment ◽  
Oxytocin Receptors ◽  
Receptor Concentration

Abstract The regulation of oxytocin, oestradiol and progesterone receptors in different uterine cell types was studied in ovariectomized ewes. Animals were pretreated with a progestogen sponge for 10 days followed by 2 days of high-dose oestradiol to simulate oestrus. They then received either low-dose oestradiol (Group E), low-dose oestradiol plus progesterone (Group P) or low-dose oestradiol, progesterone and oxytocin (via osmotic minipump; Group OT). Animals (three to six per time-point) were killed following ovariectomy (Group OVX), at oestrus (Group O) or following 8, 10, 12 or 14 days of E, P or OT treatment. In a final group, oxytocin was withdrawn on day 12 and ewes were killed on day 14 (Group OTW). Oxytocin receptor concentrations and localization in the endometrium and myometrium were measured by radioreceptor assay, in situ hybridization and autoradiography with the iodinated oxytocin receptor antagonist d(CH2)5[Tyr(Me)2,Thr4,Tyr-NH29]-vasotocin. Oestradiol and progesterone receptors were localized by immunocytochemistry. Oxytocin receptors were present in the luminal epithelium and superficial glands of ovariectomized ewes. In Group O, endometrial oxytocin receptor concentrations were high (1346 ± 379 fmol [3H]oxytocin bound mg protein−1) and receptors were also located in the deep glands and caruncular stroma in a pattern resembling that found at natural oestrus. Continuing low-dose oestradiol was unable to sustain high endometrial oxytocin receptor concentrations with values decreasing significantly to 140 ± 20 fmol mg protein−1 (P<0·01), localized to the luminal epithelium and caruncular stroma but not the glands. Progesterone treatment initially abolished all oxytocin receptors with none present on days 8 or 10. They reappeared in the luminal epithelium only between days 12 and 14 to give an overall concentration of 306 ± 50 fmol mg protein−1. Oxytocin treatment caused a small increase in oxytocin receptor concentration in the luminal epithelium on days 8 and 10 (20 ± 4 in Group P and 107 ± 35 fmol mg protein−1 in Group OT, P<0·01) but the rise on day 14 was not affected (267 ± 82 in Group OT and 411 ± 120 fmol mg protein−1 in Group OTW). In contrast, oestradiol treatment was able to sustain myometrial oxytocin receptors (635 ± 277 fmol mg protein−1 in Group O and 255 ± 36 in Group E) and there was no increase over time in Groups P, OT and OTW with values of 61 ± 18, 88 ± 53 and 114 ± 76 fmol mg protein−1 respectively (combined values for days 8–14). Oestradiol receptor concentrations were high in all uterine regions in Group O. This pattern and concentration was maintained in Group E. In all progesterone-treated ewes, oestradiol receptor concentrations were lower in all regions at all time-points. The only time-related change occurred in the luminal epithelium in which oestradiol receptors were undetectable on day 8 but developed by day 10 of progesterone treatment. Progesterone receptors were present at moderate concentrations in the deep glands, caruncular stroma, deep stroma and myometrium in Group O. Oestradiol increased progesterone receptors in the luminal epithelium, superficial glands, deep stroma and myometrium. Progesterone caused the loss of its own receptor from the luminal epithelium and superficial glands and decreased its receptor concentration in the deep stroma and myometrium at all time-points. There was a time-related loss of progesterone receptors from the deep glands of progesterone-treated ewes between days 8 and 14. These results show differences in the regulation of receptors between uterine regions. In particular, loss of the negative inhibition by progesterone on the oxytocin receptor by day 14 occurred only in the luminal epithelium, but is unlikely to be a direct effect of progesterone as no progesterone receptors were present on luminal epithelial cells between days 8 and 14. The presence of oxytocin receptors in the luminal epithelium of ovariectomized ewes suggests that oestradiol is not essential for oxytocin receptor synthesis at this site. Oestradiol was able to sustain its own receptor at all sites, but high circulating progesterone was always inhibitory to oestradiol receptors. In general, oestradiol stimulated progesterone receptors in epithelial cells whereas progesterone abolished its own receptor from epithelial cells over a period of time, but had a lesser effect on stromal cells. The concentration of all three receptors is therefore differentially regulated between different uterine cell types, suggesting the importance of paracrine effects which remain to be elucidated. Journal of Endocrinology (1996) 151, 375–393

Effects of pregnancy and systemic or intrauterine oxytocin infusion on the distribution of endometrial oxytocin receptors in the ewe: an autoradiographic study

1993 ◽  
Vol 137 (3) ◽  
pp. 423-NP ◽  
Author(s):  
V. J. Ayad ◽  
T. J. Parkinson ◽  
E. L. Matthews ◽  
M. L. Wild
Keyword(s):  
Epithelial Cells ◽  
Stromal Cells ◽  
Autoradiographic Study ◽  
Oxytocin Receptor ◽  
Oestrous Cycle ◽  
Oxytocin Receptors ◽  
Receptor Concentration ◽  
Oxytocin Infusion ◽  
Luminal Epithelial Cells ◽  
Specific Labelling

ABSTRACT Previous autoradiographic studies have suggested that the regulation of oxytocin receptors differs between endometrial cell types during the ovine oestrous cycle, and that those present on luminal epithelial cells are of particular importance to the regulation of prostaglandin F2α release during luteal regression. The present autoradiographic study compares the distribution of the endometrial oxytocin receptor in day-15 non-pregnant and pregnant ewes. The distribution of the endometrial oxytocin receptor in day-15 non-pregnant ewes infused with systemic or intrauterine oxytocin has also been investigated. Continuous, s.c. infusion of oxytocin (150 mmol/24 h) into ewes (n = 6) between days 10 and 15 of the oestrous cycle significantly increased plasma oxytocin concentrations (to approximately 100 pmol/l). There was no similar increase in systemic oxytocin concentrations in ewes receiving intrauterine (i.u.) oxytocin infusions (10 nmol/24 h) between days 10 and 15 of the oestrous cycle (n = 6). Luteolysis was inhibited in all six animals infused with oxytocin (s.c.) and endometrial oxytocin receptor concentrations were significantly lower on day 15 in these animals (12·8 ± 6·5 (s.e.m.) fmol/mg protein; P<0·001) and in pregnant ewes (18·4 ± 15·4 fmol/mg protein; P <0·001; n = 8) than in ewes infused with saline (248·6±67·1 fmol/mg protein; n = 6). While the 125I-labelled oxytocin receptor antagonist, [1-(β-mercapto-β,β-cyclopentamethylene propionic acid), 2-(ortho-methyl)-Tyr2, Thr4, Orn8, Tyr9-NH2]-vasotocin (125I-labelled OTA) clearly labelled glandular epithelia, luminal epithelium and caruncular stromal cells specifically on day 15 in saline (s.c.)-infused ewes, such specific labelling appeared to be reduced or absent from pregnant animals and those infused with oxytocin (s.c.). A significant reduction in the density of labelling of caruncular stroma (P < 0·05) and luminal epithelium (P < 0·001) was confirmed using quantitative densitometric analysis. The reduction in the labelling of endometrium in oxytocin-infused ewes was not caused by the binding of exogenous oxytocin to endometrial binding sites. Oxytocin infusion (i.u.) did not inhibit luteolysis, nor was there any significant difference in the endometrial oxytocin receptor concentration in this group of ewes on day 15 compared with those infused with saline (i.u.). There was also clear specific labelling of luminal epithelial cells with 125I-labelled OTA in ewes receiving oxytocin infused i.u. and quantification of autoradiograms failed to differentiate between these animals and those infused with saline (i.u.). It was concluded that systemic oxytocin infusion and the early establishment of pregnancy led to a clear reduction in the endometrial oxytocin receptor concentration on luminal epithelial cells, glandular epithelial cells and caruncular stromal cells, but that i.u. oxytocin infusions did not affect any of these receptor populations and notably not the luminal epithelial oxytocin receptor. The results support the contention that the luminal epithelial oxytocin receptor is involved in the luteolytic process. Journal of Endocrinology (1993) 137, 423–431

Autoradiographic localization of oxytocin receptors in the endometrium during the oestrous cycle of the ewe

1991 ◽  
Vol 130 (2) ◽  
pp. 199-NP ◽  
Author(s):  
V. J. Ayad ◽  
E. L. Matthews ◽  
D. C. Wathes ◽  
T. J. Parkinson ◽  
M. L. Wild
Keyword(s):  
Epithelial Cells ◽  
Stromal Cells ◽  
Oxytocin Receptor ◽  
Oestrous Cycle ◽  
Luminal Epithelium ◽  
Tissue Samples ◽  
Luteal Regression ◽  
Oxytocin Receptors ◽  
Secretory Glands ◽  
Luminal Epithelial Cells

ABSTRACT The present study was designed to determine the localization of the endometrial oxytocin receptor during the ovine oestrous cycle, particularly on day 14, the time of initiation of luteal regression in the ewe. Samples were obtained from 29 ewes at different stages of the oestrous cycle (several during the luteal phase and on every day between day 14 (− 2) and day + 3 of the oestrous period). Oxytocin receptors were localized autoradiographically in sections of uterine tissue, using the 125I-labelled oxytocin receptor antagonist [1-(β-mercapto-β,β-cyclopentamethylene propionic acid), 2-(ortho-methyl)-Tyr2,Thr4,Orn8,Tyr9-NH2]-vasotocin (125I-labelled OTA). There was some variation in the pattern of 125I-labelled OTA labelling between different uterine tissue samples from the same ewe and also between samples obtained from different ewes thought to be at the same stage of the oestrous cycle. A clear overall pattern did, however, emerge with 125I-labelled OTA-binding sites distributed between luminal epithelial cells, glandular epithelial cells and caruncular stromal cells to varying extents on different days of the cycle. During the luteal phase (days 5–12) clear specific labelling of endometrial tissue was generally absent. On day 14 labelling was evident on the luminal epithelium, but only in nine tissue samples out of a total of 18 studied, indicating that the entire luminal surface did not contain oxytocin receptors at this time. Between the day before oestrus and day 3 of the oestrous cycle the luminal epithelium was consistently labelled. The most extensive labelling of the remaining endometrial tissue was observed on the day of oestrus, with 125I-labelled OTA-binding sites clearly present on the stromal cells within caruncles and on a large proportion of secretory epithelia. This contrasted with the day before and the day after oestrus when labelling of glandular tissue was confined to the superficial endometrium, and labelling of caruncular stromal cells, although sometimes evident, was never as intense as on day 0. On days 2 and 3 labelling varied between being similar to that found on day 1 and being confined to the luminal epithelium and very few superficial secretory glands. The results of this study lead us to conclude that the oxytocin receptor shows a differential distribution between stromal cells, epithelial cells lining secretory glands and luminal epithelial cells during the oestrous cycle; that the steroidal regulation of the oxytocin receptor differs between endometrial cell types; and that control of the luminal epithelial oxytocin receptors is probably of particular importance to the regulation of prostaglandin F2α release at luteal regression and during the maternal recognition of pregnancy. Journal of Endocrinology (1991) 130, 199–206

Localization of oxytocin receptor mRNA in the ovine uterus during the oestrous cycle and early pregnancy

1994 ◽  
Vol 12 (1) ◽  
pp. 93-105 ◽  
Author(s):  
K R Stevenson ◽  
P R Riley ◽  
H J Stewart ◽  
A P F Flint ◽  
D C Wathes
Keyword(s):  
Mrna Expression ◽  
Optical Density ◽  
Receptor Gene ◽  
Oxytocin Receptor ◽  
Oestrous Cycle ◽  
Luminal Epithelium ◽  
Binding Studies ◽  
Receptor Mrna ◽  
Oxytocin Receptors ◽  
Ovine Uterus

ABSTRACT A synthetic 45-mer oligonucleotide corresponding to part of the ovine endometrial oxytocin receptor cDNA was hybridized to sections of ovine uterus collected from 40 ewes at different stages during the oestrous cycle, the first 3 weeks of pregnancy and seasonal anoestrus. The quantity of oxytocin receptor mRNA was measured as the optical density (OD) value on autoradiographs using image analysis. Message first appeared in the luminal epithelium on days 14–15 of the cycle, increasing to a peak OD of 0·48 at oestrus and decreasing again between days 2 and 5. Oxytocin receptor mRNA in the superficial glands, deep glands and caruncular stroma increased between day 15 and oestrus to peak OD values of 0·17, 0·11 and 0·11 respectively, declining again by day 2 and reaching basal values (OD<0·015) by day 5. Hybridization to the myometrium tended to rise from a mean OD value of 0·01 on days 2–15 to a peak of 0·03±0·01 (mean±s.e.m.) on days 0–1, but the change was not significant. In pregnant ewes there was no detectable oxytocin receptor mRNA on days 14–15 in any region, but hybridization to the luminal epithelium was present in two of three ewes on day 21. In anoestrous ewes oxytocin receptor mRNA concentrations in all areas of the endometrium were approximately half those measured at oestrus. Optical density readings for oxytocin receptor mRNA in the various uterine compartments were compared with measurements of oxytocin receptors in the same regions as assessed by binding studies using the 125I-labelled oxytocin antagonist d(CH2)5[Tyr(Me)2,Thr4,Tyr-NH29]-vasotocin (125I-labelled OTA). In the endometrium, receptor mRNA and 125I-labelled OTA binding patterns changed in parallel, and both sets of measurements were significantly correlated (P<0·01). In the myometrium, a significant increase in 125I-labelled OTA binding occurred at oestrus; this was not accompanied by a similar increase in oxytocin receptor mRNA hybridization. This study helps to confirm that the previously identified cDNA clone is derived from the ovine oxytocin receptor, as patterns of oxytocin receptor mRNA expression in the endometrium closely resembled those of oxytocin binding. Maximum expression and binding both occurred at oestrus, suggesting that regulation of the oxytocin receptor gene in the uterus occurs principally at the transcriptional, rather than at the translational, level. Failure to detect a significant increase in myometrial mRNA expression at oestrus may indicate that the endometrial and myometrial oxytocin receptors are of different isoforms.

scholarly journals Gene-expression changes induced by Feline immunodeficiency virus infection differ in epithelial cells and lymphocytes

2005 ◽  
Vol 86 (8) ◽  
pp. 2239-2248 ◽  
Author(s):  
R. J. O. Dowling ◽  
D. Bienzle
Keyword(s):  
Gene Expression ◽  
Epithelial Cells ◽  
Virus Infection ◽  
Feline Immunodeficiency Virus ◽  
Cell Types ◽  
Early Gene ◽  
Time Points ◽  
Subtype B ◽  
Immunodeficiency Virus ◽  
Transcriptional Changes

Infection of cats with Feline immunodeficiency virus (FIV) is an important model for understanding comparative lentivirus biology. In vivo, FIV infects lymphocytes and monocyte/macrophages, but in vitro infection is commonly investigated in epithelial Crandell–Reese Feline Kidney (CRFK) cells. In this study, the transcriptional responses of CRFK cells and primary lymphocytes to infection with FIV 34TF, a cloned subtype A virus, and FIV USgaB01, a biological subtype B isolate, were determined. Reverse-transcribed mRNA from both cell types was hybridized to microarrays containing 1700 human expressed sequence tags in duplicate and data were analysed with Significance Analysis of Microarrays (sam) software. Results from six experiments assessing homeostatic cross-species hybridization excluded 3·48 % inconsistently detected transcripts. Analysis of data from five time points over 48 h after infection identified 132 and 24 differentially expressed genes in epithelial cells and lymphocytes, respectively. Genes involved in protein synthesis, the cell cycle, structure and metabolism were affected. The magnitude of gene-expression changes ranged from 0·62 to 1·62 and early gene induction was followed by downregulation after 4 h. Transcriptional changes in CRFK cells were distinct from those in lymphocytes, except for heat-shock cognate protein 71, which was induced at multiple time points in both cell types. These findings indicate that FIV infection induces transcriptional changes of a modest magnitude in a wide range of genes, which is probably reflective of the relatively non-cytopathic nature of virus infection.

scholarly journals Salmonella enters a dormant state within human epithelial cells for persistent infection

2021 ◽  
Vol 17 (4) ◽  
pp. e1009550
Author(s):  
Chak Hon Luk ◽  
Camila Valenzuela ◽  
Magdalena Gil ◽  
Léa Swistak ◽  
Perrine Bomme ◽  
...  
Keyword(s):  
Epithelial Cells ◽  
Stringent Response ◽  
Cell Types ◽  
Enteric Bacterium ◽  
Fluorescent Reporter ◽  
Dormant State ◽  
Time Points ◽  
Wide Range ◽  
Effector Secretion ◽  
Vesicular Compartment

Salmonella Typhimurium (S. Typhimurium) is an enteric bacterium capable of invading a wide range of hosts, including rodents and humans. It targets different host cell types showing different intracellular lifestyles. S. Typhimurium colonizes different intracellular niches and is able to either actively divide at various rates or remain dormant to persist. A comprehensive tool to determine these distinct S. Typhimurium lifestyles remains lacking. Here we developed a novel fluorescent reporter, Salmonella Intracellular Analyzer (SINA), compatible for fluorescence microscopy and flow cytometry in single-bacterium level quantification. This identified a S. Typhimurium subpopulation in infected epithelial cells that exhibits a unique phenotype in comparison to the previously documented vacuolar or cytosolic S. Typhimurium. This subpopulation entered a dormant state in a vesicular compartment distinct from the conventional Salmonella-containing vacuoles (SCV) as well as the previously reported niche of dormant S. Typhimurium in macrophages. The dormant S. Typhimurium inside enterocytes were viable and expressed Salmonella Pathogenicity Island 2 (SPI-2) virulence factors at later time points. We found that the formation of these dormant S. Typhimurium is not triggered by the loss of SPI-2 effector secretion but it is regulated by (p)ppGpp-mediated stringent response through RelA and SpoT. We predict that intraepithelial dormant S. Typhimurium represents an important pathogen niche and provides an alternative strategy for S. Typhimurium pathogenicity and its persistence.

scholarly journals [Ciliogenesis in the mucous cells of the quail oviduct. II. Hormonal control]

1976 ◽  
Vol 71 (2) ◽  
pp. 460-471 ◽  
Author(s):  
D Sandoz ◽  
E Biosvieux-Ulrich ◽  
C Laugier ◽  
E Brard
Keyword(s):  
Epithelial Cells ◽  
Estradiol Benzoate ◽  
Hormonal Control ◽  
Secretory Cells ◽  
Luminal Epithelium ◽  
Mucous Cells ◽  
Ciliated Cells ◽  
Cell Divisions ◽  
Progesterone Treatment

The hormonal control of ciliogenesis and transformation of mucous cells was studied in the oviduct (magnum) of ovariectomized quails. Estradiol benzoate induces ciliogenesis with doses varying from 10 mug/day to 100 mug/day after 6 days of treatment. With 100 mug/day, differentiation of some mucous cells is also induced as well as the formation of transitory "mixed cells" which are in the process of ciliogenesis and contain mucous granules. Associated with progesterone (1 mg/day), estradiol benzoate (10 mug/day) induces the differentiation of mucous cells and ciliated cells. The luminal epithelium of quails injected with this mixture is similar to the luminal epithelium observed in the oviduct of laying quails. With the same dose of progesterone (1 mg/day) and 20 mug/day of estradiol benzoate for 6 days, ciliogenesis is completely inhibited. All epithelial cells are secretory cells. Transformation of 50% of the mucous cells into ciliated cells is obtained by following the previous estradiol-progesterone treatment with the injection of estradiol benzoate (20 mug/day) for 3 days. Divisions of mucous cells were also observed. It is also possible to induce ciliogenesis in some mucous cells by withdrawing both hormones for 3 days. In this case, no cell divisions were observed.

Regulation of oxytocin receptor concentration in rat uterine explants by estrogen and progesterone

1983 ◽  
Vol 61 (7) ◽  
pp. 625-630 ◽  
Author(s):  
Melvyn S. Soloff ◽  
Mats Axel Fernstrom ◽  
Sumudra Periyasamy ◽  
Solweig Soloff ◽  
Sam Baldwin ◽  
...  
Keyword(s):  
Protein Synthesis ◽  
Oxytocin Receptor ◽  
Fivefold Increase ◽  
Oxytocin Receptors ◽  
Immature Rats ◽  
Receptor Concentration ◽  
17Β Estradiol ◽  
Inhibitor Of Protein Synthesis

Incubation of uterine explants from immature rats with 0.01–100 ng of 17β-estradiol/mL resulted in approximately a fivefold increase in the number of oxytocin receptors per milligram of protein in 48 h. This increase was maintained for at least an additional 48 h in the presence of estrogen. When the explants were incubated with 1 μg progesterone/mL from the outset, the concentration of oxytocin receptors was the same as initial (0 time) levels. The estrogen-induced increase in oxytocin receptor concentration was blocked by incubation with cycloheximide, an inhibitor of protein synthesis. Once increased, however, the concentration of oxytocin receptors exhibited no turnover for at least a 48-h period in the presence of estrogen. The addition of progesterone and estrogen to explants with elevated receptor levels resulted in almost a 60% reduction in oxytocin receptor concentration by 24 h, with no change in affinity of the receptor for oxytocin. The reduction in receptor concentration by progesterone was not prevented by cycloheximide. The progesterone effect may involve inactivation or degradation of oxytocin receptors or activation of substances that are inhibitory to oxytocin binding. The effects of estradiol and progesterone on oxytocin receptor concentration in uterine explants are similar to those seen when the steroids are administered in vivo. The explant system, therefore, should prove useful in clarifying factors and processes that are involved in regulation of oxytocin receptor concentration in the uterus and in the initiation of parturition in the rat.

Ovine conceptus secretory proteins and bovine recombinant interferon α1-1 decrease endometrial oxytocin receptor concentrations in cyclic and progesterone-treated ovariectomized ewes

1991 ◽  
Vol 131 (3) ◽  
pp. 475-482 ◽  
Author(s):  
J. L. Vallet ◽  
G. E. Lamming
Keyword(s):  
Serum Proteins ◽  
Oxytocin Receptor ◽  
Placebo Control ◽  
Interferon Α ◽  
Secretory Proteins ◽  
Recombinant Interferon ◽  
Blood Samples ◽  
Progesterone Treatment ◽  
Oxytocin Receptors ◽  
Prostaglandin F

ABSTRACT A series of experiments was performed to determine whether proteins produced by the sheep conceptus (oCSP) during the time of maternal recognition of pregnancy or bovine recombinant interferon α1-1 (brIFN) decrease oxytocin receptor concentrations in the endometrium of cyclic or ovariectomized progesterone-treated ewes. In experiment 1, cyclic ewes received intrauterine infusions of serum proteins (oSP), oCSP or brIFN on days 12, 13 and 14 of the oestrous cycle. Ewes then received an oxytocin challenge (1 μg in 0·9% NaCl), and blood samples were taken just before and every 10 min for 1 h after the challenge; these were measured for 13,14-dihydro-15-ketoprostaglandin F2α (PGFM), the stable metabolite of prostaglandin F2α. Endometrial oxytocin receptor concentrations were then measured. The oCSP and brIFN treatments suppressed both endometrial oxytocin receptor concentrations and oxytocin-induced increases in PGFM concentrations. In experiment 2, ewes were ovariectomized and then pretreated with a fluorogestone acetate-releasing intravaginal device for 10 days followed by oestradiol (25 μg i.m. twice daily for 2 days). Ewes were then treated with progesterone (10 mg i.m. twice daily for 12 days). Ewes received intrauterine infusions of oSP, oCSP and brIFN on days 10, 11 and 12 of progesterone treatment. On the day after the last progesterone treatment, ewes were challenged with oxytocin and blood samples collected to measure PGFM. Endometrial oxytocin receptors were also measured. Treatment with oCSP, but not brIFN, suppressed endometrial concentrations of oxytocin receptor, and neither oCSP nor brIFN altered oxytocin-induced increases in PGFM concentrations. In experiment 3, ewes were ovariectomized and pretreated as in experiment 2 and then received progesterone treatment for 6, 8, 10 or 30 days. On the day after the last progesterone treatment, ewes received an oxytocin challenge and blood samples and endometrium were collected as in experiment 1. Endometrial oxytocin receptors increased sharply between days 8 and 10 and remained raised after 30 days of progesterone treatment. Oxytocin-induced PGFM increased between 8 and 10 days of progesterone treatment, but no response to oxytocin was detected after 30 of progesterone treatment. In experiment 4, ewes were pretreated as in experiment 2 and then treated for 10 days with progesterone and received intrauterine infusions of oCSP, oSP or brIFN placebo control buffer on days 8, 9 and 10. Ewes received oxytocin and blood samples and endometrium were collected as in experiment 1. As in experiment 2, oCSP treatment suppressed oxytocin receptor concentrations but did not affect oxytocin-induced PGFM release. In experiment 5, ewes were treated with steroid hormones as in experiment 4 and then received intrauterine infusions of either brIFN or oSP on days 8, 9 and 10 of progesterone treatment. The brIFN treatment suppressed oxytocin receptor concentrations but did not suppress oxytocin-induced increases in plasma PGFM. We concluded from these experiments that (1) treatment of cyclic ewes with either oCSP or brIFN decreases endometrial oxytocin receptor concentrations and oxytocin-induced increases in PGFM and (2) in progesterone-treated ovariectomized ewes, treatment with oCSP and brIFN suppresses endometrial oxytocin receptor concentrations but does not suppress oxytocin-induced increases in PGFM. Journal of Endocrinology (1991) 131, 475–482

Localization of oestradiol, progesterone and oxytocin receptors in the uterus during the oestrous cycle and early pregnancy of the ewe

1993 ◽  
Vol 138 (3) ◽  
pp. 479-NP ◽  
Author(s):  
D. C. Wathes ◽  
M. Hamon
Keyword(s):  
Progesterone Receptor ◽  
Early Pregnancy ◽  
Luteal Phase ◽  
Specific Binding ◽  
Progesterone Receptors ◽  
Oestrous Cycle ◽  
Luminal Epithelium ◽  
Paracrine Factors ◽  
Plasma Progesterone ◽  
Oxytocin Receptors

ABSTRACT Uterine tissue samples were collected from 47 ewes at various stages of the oestrous cycle and early pregnancy (until day 21) and during seasonal anoestrus. Cryostat sections were immunostained to determine the localization of oestradiol and progesterone receptors using specific monoclonal antibodies. Oxytocin receptors were localized by autoradiography in sections from the same ewes using the 125I-labelled oxytocin antagonist d(CH2)5[Tyr(Me)2,Thr4,Tyr-NH29]- vasotocin. Plasma progesterone measurements were made during the preceding cycle up to the time of slaughter. Oestradiol receptor concentrations were maximal in all regions of the tract at oestrus. Immunostaining of the luminal epithelium, superficial glandular epithelium, stroma and myometrium decreased in the early luteal phase but was maintained for longer in the deep glands. Progesterone receptor immunostaining in the luminal epithelium and superficial glands developed in the early luteal phase (days 1–2) with a somewhat later appearance in the deep glands (days 5–7). Progesterone receptor concentrations in the stroma and myometrium also reached a maximum in the early luteal phase. Myometrial staining was clearly maintained throughout the luteal phase whereas stromal staining was variable between ewes. For both oestradiol and progesterone receptors no differences were apparent between pregnant and non-pregnant ewes between days 2 and 12, but pregnant ewes did not show the general increases in oestradiol receptor staining associated with luteolysis on days 14–15. Oxytocin receptors first developed in the luminal epithelium of non-pregnant ewes on day 14 of the cycle and spread to the superficial glands, caruncular stroma, deep glands and myometrium at oestrus before decreasing in reverse order on days 1–2. Specific binding was not detectable on days 5–12 of the cycle or on days 14 or 21 of pregnancy. The appearance of oxytocin receptors in the luminal epithelium on day 14 preceded that of both the oestradiol and progesterone receptors in the epithelial cells and the fall in plasma progesterone. It was followed by the development of oestradiol and oxytocin receptors in the superficial glands, deep glands, caruncular stroma and myometrium, with the two receptor populations showing a significant positive association in these tissues. The loss of oxytocin receptors in all regions occurred as plasma progesterone levels were increasing, but the association between these two variables was only significant in the superficial glands. The development of progesterone receptors in different tissues could not be explained on the basis of either oestradiol receptor content or plasma progesterone. We conclude that all three receptor populations change in a dynamic manner during the oestrous cycle with variations both between days and between different uterine compartments. The complex pattern of receptor formation and loss suggests that, in addition to the circulating steroid hormone concentrations, local paracrine factors are likely to be involved in their regulation. Journal of Endocrinology (1993) 138, 479–491

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