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P. A. Denning-Kendall
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M. L. Wild
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Wathes D. C.
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ABSTRACT

Bovine corpora lutea and ovarian stroma were analysed by high-performance liquid chromatography for catecholamine content. High concentrations (up to 102 nmol/g wet weight) were found in both 'central' stroma, containing many blood vessels, and 'peripheral' stroma. Central stroma contained noradrenaline and some dopamine, whereas peripheral stroma contained a higher proportion of dopamine and also significant amounts of 3,4-dihydroxyphenylacetic acid (DOPAC). Occasional samples of stroma had very high amounts of dopamine, suggesting that it is stored in specific regions. Corpora lutea, although devoid of direct innervation, contained dopamine (up to 5·3 nmol/g) and noradrenaline (up to 1·2 nmol/g). The average dopamine: noradrenaline molar ratio was 1·19 : 1 and the concentrations of dopamine and noradrenaline were highly correlated (P < 0·002). The concentration of dopamine was significantly higher in the early luteal phase of the oestrous cycle than during the rest of the cycle or in pregnancy. The levels of noradrenaline and dopamine present in corpora lutea are sufficient to modulate the production of both oxytocin and progesterone by luteal cells in vitro.

Journal of Endocrinology (1991) 129, 221–226

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V. J. Ayad
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T. J. Parkinson
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E. L. Matthews
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M. L. Wild
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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 F 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

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V. J. Ayad
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E. L. Matthews
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D. C. Wathes
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T. J. Parkinson
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M. L. Wild
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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 F release at luteal regression and during the maternal recognition of pregnancy.

Journal of Endocrinology (1991) 130, 199–206

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