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Department of Pharmacology, Materia Medica and Therapeutics, Medical School, University of Manchester, Oxford Road, Manchester, M13 9PT
(Received 15 October 1976)
The intra-uterine balloon is one of the commonest methods used in investigating uterine contractility. The micro-balloon measures the maximum intra-uterine pressure that is developed in a closed system as the myometrium contracts. That intra-uterine pressure measurement is a reliable reflection of myometrial contractility at a physiological level was shown by Csapo (1963) who recorded the electrical activity of the uterus simultaneously with intra-uterine pressure and found good correlation between the two in the rabbit. The micro-balloon method was adopted to study various aspects of the pharmacological control of uterine activity in late gestation in the rat by means of various anti-enzymes. Fuchs (1969) has used large intra-uterine balloons and Boer, Lincoln & Swaab (1975) have used pressure sensitive pills to record uterine activity during late pregnancy and parturition in
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Search for other papers by L A Salamonsen in
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Abstract
Previous studies have demonstrated that endothelin is present in the ovine endometrium and increases at around the expected time of implantation. To characterize further uterine endothelin at the time of establishment of pregnancy in sheep, endothelin was measured by radioimmunoassay in uterine flushings obtained during the oestrous cycle and in pregnant ewes up to the time of implantation (day 16). During the oestrous cycle, the highest amounts of endothelin were present in uterine flushings on day 14 (1·1 ±0·2 ng endothelin/uterus). During early pregnancy, basal levels of endothelin (0·5–0·6 ng endothelin/uterus) were present in uterine flushings for the first 10 days and then increased on day 14 to levels similar to those found at the equivalent stage of the oestrous cycle. On days 15 and 16 of pregnancy, endothelin content in the uterine lumen increased to significantly (P<0·05) higher concentrations (2·9±0·4 ng endothelin/uterus) when compared with the non-fertile cycle. The principal isoform present in flushings at the time of implantation was endothelin-1, as determined by reverse-phase HPLC. Endothelin was released principally by purified endometrial epithelial cells in culture, with barely detectable amounts released by endometrial stromal cells or conceptus tissue, which is consistent with the epithelium being the principal source of endothelin in the uterine lumen. Endothelin binding sites were present in endometrium and myometrium, as demonstrated by specific binding of 125I-labelled endothelin-1, which was saturable and displaced by endothelin-1. Both endothelinA and B sub-types of receptors were present as demonstrated by the biphasic displacement of 125I-labelled endothelin-1 binding by the specific endothelinB agonist BQ3020. These were localised principally on luminal and glandular epithelium and in the vasculature of the endometrium and myometrium as shown by autoradiography. Endothelin receptors were also present on the conceptus obtained at the time of implantation. In the day 20 conceptus, endothelin immunostaining was localised principally in the heart, in trophoblast in uninucleate but not in binucleate cells, and in fetal membranes. This immunostaining of the conceptus may represent binding to receptor sites. It is concluded that endothelin-1 is present in the uterine lumen and may play an important role in the paracrine regulation of the conceptus and endometrium at the time of rapid embryo development, implantation and early placentation.
Journal of Endocrinology (1995) 147, 235–244
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Abstract
The entire coding region of an ovine endometrial oxytocin receptor (OTR) cDNA was generated by PCR, subcloned into the SV40 major late promoter expression vector pSVLJ and transiently expressed in Cos-7 cells. A specific OTR antagonist, 125I-labelled d(CH2)5 [Tyr(Me)2,Thr4,Tyr-NH2 9]-vasotocin (OTA), was used to describe the binding kinetics of the expressed receptor which had a K d of 4·5 nm and Bmax of 2·4 nm/mg protein (6·8 × 105 receptor molecules/transfected cell). The functional properties of the expressed OTR were determined by measuring oxytocin-induced phosphoinositide (PI) hydrolysis. Oxytocin increased PI turnover in OTR transfected cells fourfold in excess of residual endogenous activity, and stimulated phospholipase C (PLC) activity in a dose- and time-dependent manner, confirming that the expressed OTR cDNA was functional. Arginine vasopressin also stimulated PI turnover in a dose-dependent manner; thresholds of responses to oxytocin and arginine vasopressin were 10−9 m and 10−7 m respectively. OTA did not increase PI turnover and competitively inhibited the oxytocin-induced response. Direct activation of the pathway by aluminium fluoride and guanosine (3′-Othio)-triphosphate (GTPγS) confirmed that the OTR was G-protein linked. Co-incubation of GTPγS with oxytocin shifted the PI-response threshold from 10−7 m to 10−9 m and significantly increased the level of response, suggesting that maximum PI turnover was agonist-dependent. The G-protein involved in mediating the signal transduction pathway was pertussis toxin-insensitive and, therefore, probably a member of the Gq subfamily. The PLC inhibitor, U73122, had no effect on oxytocin-induced PI turnover, consistent with the response in endometrial tissue. These data suggest that the signalling pathway mediated by expressed OTR is similar to that attributed to OTR occupancy in ovine endometrium.
Journal of Endocrinology (1996) 149, 389–396
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SUMMARY
The concentrations of peripheral plasma testosterone were measured by radioimmunoassay in samples collected from five bulls, each given i.m. injections of 0, 5, 15, 30 and 60 mg prostaglandin F2α (PGF2α). Synchronized peaks in testosterone concentration occurred with maximum values 1–2 h after treatment. These increases of testosterone persisted significantly longer than those observed to occur as natural episodic peaks during two 24 h periods in the same bulls. The mean peak testosterone concentration after PGF2α injection was related to the dose of PGF2α, values after 60 and 30 mg doses being significantly greater than after 15 mg. The response produced by a 5 mg dose was not significant. The results indicate that intramuscular injection of PGF2α acutely stimulates synthesis and release of testosterone in bulls.