ABSTRACT

It has previously been shown that administration of the indoleamine melatonin to advance the breeding season of ewes is also associated with an increase in ovulation rate and subsequent litter size.

Experiment 1 assessed whether, in ewes receiving melatonin to advance the breeding season, the indoleamine acts directly on the corpus luteum to enhance progesterone secretion or indirectly through increased activity of the hypothalamic pulse generator. Six ewes received 3 mg melatonin orally at 15.00 h daily from 22 March onwards, six were induced to ovulate during mid-anoestrus following withdrawal of a progestagen pessary and injection of exogenous gonadotrophin and six acted as naturally ovulating controls. First overt oestrus occurred between 17 May and 8 July in melatonin-treated ewes, between 21 October and 3 January in control ewes and on 8 July in all induced ewes. On days 2 and 10 after the first overt oestrus, melatonin-treated ewes had pulsatile LH activity characteristic of that measured in control ewes ovulating naturally during the breeding season. There was an absence of any pulsatile LH activity in the induced ewes. Progesterone concentrations between days 7 and 12 following oestrus were significantly higher in melatonin-treated than in control and induced ewes, suggesting a luteotrophic role for melatonin.

Experiment 2 was carried out to determine whether administration of melatonin commencing after induced ovulation and insemination would alter the endocrine status of the ewe and thereby influence the establishment of pregnancy and embryo survival. Thirty-two anoestrous ewes were induced to ovulate on 29 June. Starting 24 h after intra-uterine insemination, 16 ewes were given melatonin daily for 60 days and 16 acted as controls. Daily LH concentrations were higher in melatonin-treated than in control ewes from days 2 to 22 after oestrus, while prolactin concentrations declined in melatonin-treated ewes over the same period. Plasma progesterone concentrations were enhanced in melatonin-treated ewes between days 4 and 9 following oestrus, yet ovulation rates were the same as for controls. Successful pregnancies occurred in 0·56 control (9 of 16) and 0·69 melatonin-treated (11 of 16) ewes. For these ewes the number of fetuses surviving to term as a proportion of ovulation rate was 0·43 and 0·51 for the control and melatonin treatment respectively.

J. Endocr. (1988) 119, 523–530

ABSTRACT

Induction of ovulation early post partum in sheep is associated with a high incidence (30–40%) of premature luteolysis. The present study was designed to characterize oxytocin receptor levels, oxytocin-stimulated inositol phosphate (IP) turnover (second messenger) and oxytocin-stimulated prostaglandin F_2α (PGF_2α) release in the endometrium of post-partum ewes induced to ovulate 21 days after parturition and expected to exhibit a range of corpus luteal functions subsequently.

Ovulation was induced on day 21 post partum using a controlled internal drug release device and pregnant mare serum gonadotrophin, and uterine tissues were collected on days 5, 10 or 15 of the cycle (n = 4/day). A further 12 ewes whose interval from previous parturition exceeded 150 days were similarly treated and acted as controls. Measurement of daily peripheral progesterone concentrations revealed that while all control ewes exhibited normal luteal function, abnormal luteal function was evident in two, two and one post-partum ewes studied on days 5, 10 and 15 of the cycle respectively. Oxytocin receptor binding was detected (by receptor-binding assay and in-vitro autoradiography) in the endometrium and myometrium of post-partum ewes at all three stages of the oestrous cycle but only at day 15 in control ewes. To determine IP turnover, 100 mg caruncular endometrium was incubated in duplicate for 2·5 h with 10 μCi [³H]inositol and treated with 0 or 2 μmol oxytocin/l for 30 min, then [³H]inositol mono-, bis- and trisphosphates were quantified. Oxytocin stimulated total IPs in all day-5 and day-15 post-partum ewes, in three of four day-10 ewes and in all day-15 control ewes. Basal endometrial PGF_2α release measured in triplicate (100 mg/well) during a 2 h incubation was higher in post-partum versus control ewes on days 5 and 10 but not on day 15 of the cycle. Similarly, oxytocin stimulated PGF_2α release to varying levels at all stages of the cycle in post-partum ewes but only on day 15 in control ewes. Irrespective of the treatment group endometrial oxytocin receptor number was significantly (P < 0·001) correlated with oxytocin-stimulated IP turnover and PGF_2α release.

Thus the induction of ovulation and the subsequent luteal phase in post-partum ewes is against a back ground of high oxytocin receptor expression and enhanced PGF_2α release which in some ewes may contribute to abnormal luteal function.

Journal of Endocrinology (1993) 136, 17–25

ABSTRACT

The induction of ovulation in early post-partum ewes is associated with a high incidence of premature luteal regression which is independent of the suckling stimulus but dependent on the stage post partum. The aim of the present study was to determine whether oxytocin receptors are present on uterine endometrium early in the luteal phase and hence ascertain whether oxytocin-induced uterine prostaglandin F_2α release is a possible mechanism involved in the premature regression of these post-partum corpora lutea. Ovarian and uterine tissues were collected on day 4 of the cycle in ewes induced to ovulate at either 21 or 35 days post partum (n = 4 per group). A further four cyclic ewes were similarly synchronized to ovulate and acted as controls. Corpora lutea from the 21-day post-partum group were significantly (P < 0·01) smaller, had a lower progesterone content and a reduced capacity to secrete progesterone in vitro than corpora lutea from 35-day post-partum or control ewes.

A highly specific oxytocin receptor ligand ¹²⁵I-labelled d(CH₂)₅[Tyr(Me)²,Thr⁴,Tyr-NH₂ ⁹]-vasotocin was used to localize and characterize high affinity oxytocin receptors in uterine endometrium (dissociation constant 145 pmol/l). Oxytocin receptor concentrations in endometrium from ewes induced to ovulate at 21 days post partum were on average five-fold higher (P < 0·05) than in 35-day post-partum and control groups.

Journal of Endocrinology (1991) 128, 253–260