A correlation between ovarian follicular maturity and anchorage-independent growth of bovine granulosa cells.

Previous studies have shown that a subset of bovine ovarian granulosa cells can proliferate to form clones of functional cells in suspension in a semisolid agar matrix, without the requirement for attachment to the substratum. These clonogenic anchorage-independent granulosa cells are responsive to epidermal growth factor and exhibit properties of a primitive progenitor cell population. We have used this assay system to analyze the proliferation of granulosa cells during ovarian follicular maturation.

Role of intrafollicular regulators and FSH in growth and development of large antral follicles in sheep.

Manipulation of circulating concentrations of hormones and ovarian follicle status was carried out on Day 11-12 of the oestrous cycle in sheep. All follicles visible on the ovary were ablated by cautery and ewes were treated with oestradiol or ovine follicular fluid (oFF) to suppress FSH or with PMSG to increase circulating gonadotrophic activity. One group underwent unilateral ovariectomy which greatly increased endogenous FSH and was the only treatment which significantly affected LH pulse frequency.

Seasonal changes in LH secretion in normal ewes and ewes which grazed oestrogenic clover.

Plasma luteinizing hormone (LH) concentrations were measured in normal (control) Corriedale X Merino (comeback) ewes and in clover-infertile comeback ewes which had grazed oestrogenic Yarloop clover (Trifolium subterraneum L. cv. Yarloop) for more than 4 years.

Relaxin inhibits the plateau component of the action potential in the circular myometrium of the rat.

The effects of relaxin on contractility and membrane potential of the longitudinal and circular smooth muscle layers of the uterus have been studied in vitro using oestrogen-treated, non-pregnant rats and pregnant rats. Relaxin decreased the amplitude of contractions induced by electrical stimulation of longitudinal myometrium by decreasing the duration of the bursts of action potentials. This effect was transient and tachyphylaxis always developed and was observed following injection of steroids and up to day 17 of pregnancy.

Differences in the reproductive endocrine status of ewes in the early post-partum period and during seasonal anoestrus.

More (P less than 0.05) post-partum acyclic ewes (8/9) showed evidence of pulsatile LH release than did seasonally anoestrous ewes (2/8). Mean plasma prolactin concentrations were higher (P less than 0.

The characteristics of an anchorage-independent clonal agar assay for primary explanted bovine granulosa cells.

Normal, primary explanted, bovine granulosa cells grow reproducibly in agar culture as anchorage-independent clones. Epidermal growth factor (EGF) and rat erythrocytes are effective stimulators of colony formation, and when both are added to the culture medium at optimal concentrations, there is an enhancement of colony numbers and colony size, indicative of an independent, and operationally additive, mode of action for the two factors. The ability of cells propagated from agar clones to secrete progesterone, and to augment progesterone secretion 4-fold in the presence of 1 mM dbcAMP is proof that colonies originate from and are composed of functional granulosa cells.

The time of oestrus and ovulation following various synchronisation techniques using progesterone impregnated intravaginal devices.

Progesterone releasing intravaginal devices (PRIDs) were used in 4 experiments involving 67 cattle to study the effect of the stage of the oestrous cycle and of oestradiol benzoate (ODB) administration on the time interval from PRID removal to oestrus and/or ovulation. Cows in which PRIDs were inserted on days 2 to 4 of the oestrous cycle for 14 days were subsequently observed by endoscopy to ovulate significantly later than cows given identical treatments on days 13 and 14 of the cycle. The concentration of progesterone was higher in the former group at the time PRIDs were removed and remained at a higher level for 3 days thereafter.

Potencies of porcine relaxins using two bioassays.

Relaxin reference preparations NIH-R-P1 and Warner Lambert W1164-3, and purified relaxin peptides, CM-a', CM-a and CM-B, were assayed in the mouse interpubic ligament and rat uterine inhibition bioassays. There was evidence that CM-a and CM-B would bind to glass and significant apparent increases in potency in the case of these two peptides alone resulted from the use of silicone-coated glassware. Using treated glassware, CM-a', CM-a and CM-B were equipotent in the mouse interpubic ligament bioassay with potencies relative to NIH-R-P1 of 3.

Hypothalamic-pituitary function in normal ewes and ewes which grazed oestrogenic subterranean clover for several years.

Plasma luteinizing hormone (LH) concentrations were measure in normal Merino ewes and in Merino ewes with lowered fertility which had resulted from prolonged grazing of Trifolium subterraneum L. cv. Dinninup.

Critical time period for the vagino-cervical stimulus to induce pseudopregnancy in the dioestrous rat.

Groups of rats experienced mechanical stimulation of the vagino-cervical region in induce pseudopregnancy at various intervals from 0900 h on dioestrus -- 1 (D -- 1) to 1800 h on D -- 2. Twenty-four of 26 rats which were stimulated on D -- 1 experienced immediate pseudopregnancy whereas when the stimulus was given on D -- 2, 39 of 45 rats continued to cycle and the latest time when such mechanical stimulation would reliably induce immediate pseudopregnancy was 2400 h on D -- 1. Abolition of the dark phase between 1800 h D -- 1 and 0600 h D -- 2 did no alter this critical time period (2400 h on D -- 1).

Pituitary responsiveness to LH-RH in post-partum ewes treated with oestradiol-17 beta and failing to show a plasma LH surge.

Oestradiol-17 beta (40 micrograms intravenously) failed to elicit a surge in plasma LH levels by 13 h after administration in 64% (16 out of 25) Merino ewes about 30 days post partum in the anoestrous season. LH-RH responsiveness and LH-RH priming effect were significantly greater in these ewes than in similar post-partum (n = 9) and non-parturient ewes (n = 3) not treated with oestradiol. This suggests that the failure of the oestrogen-positive feedback effect on LH release in post-partum ewes is not due to a failure of oestradiol action on the pituitary increasing pituitary responsiveness to LH-RH and LH-RH priming effect, but could be due to inadequate release of LH-RH from the hypothalamus.

Identification of relaxins in porcine follicular fluid and in the ovary of the immature sow.

Relaxin was measured by radioimmunoassay in follicular fluid from sows which were of different reproductive states. In non-pregnant animals, follicular fluid relaxin values ranged from 0.23 to 666 ng/ml.

Plasma relaxin immunoactivity in the pig at parturition and during nuzzling and suckling.

One sow bled at 30--60-min intervals for 48 h at 5 and 4 days before parturition had mean +/- s.e.m.

Pituitary response to LHRH during the puerperium in women.

Five breast-feeding women received 100 mug synthetic LHRH at between 19 and 58 days post-partum. One subject was treated twice. LH and FSH responses were monitored using radioimmunoassay.

Gonadotrophin levels and ovarian development in the neonatal ewe lamb.

Ovarian weight has been shown to increase markedly in the newborn ewe lamb, and factors which might contribute to this growth were examined. Follicle development was studied in the ovaries of 28 lambs aged 0, 2, 4, 6, 8 and 10 weeks. Plasma samples, pituitaries and one ovary were assayed for gonadotrophin or steroid hormone content.

Prolactin response to synthetic TRF in pregnant ewes.

The prolactin secretion response to TRF was similar at all stages of pregnancy (Days 40, 66, 102 and term).

Effect of prolactin on the LH response to synthetic LH-RH in ovariectomized ewes.

When ovariectomized ewes were treated with LH-RH, all 3 receiving prolactin infusion and 4 out of 5 receiving an infusion of NaCl solution responded.

The effect of testosterone and an anti-oestrogen on hypertrophy of bulbourethral glands of wethers grazing oestrogenic pastures.

Testosterone implants (70 mg), but not an oral anti-oestrogen, prevented hypertrophy of the bulbourethral glands of wethers grazing oestrogenic pastures for 18 weeks.

In vitro response of relaxin-treated rat uterus to prostaglandins and oxytocin.

Paired segments of rat uterus were treated in vitro with relaxin (W1164-3, 150 GPU/mg) until the amplitude of contraction was reduced to at least 50% of the pre-treatment amplitude. Test segments then received 100 ng of either PGE1, PGE2, PGF2alpha or 250 uU of oxytocin. Control segments remained untreated.

Content of LH, FSH, and growth hormone in the pituitaries of pregnant and anestrous sheep.

The total content of LH, FSH, and growth hormone was measured in pituitaries from anestrous ewes and from ewes at known stages of pregnancy. LH content was lower in the pregnant ewes than in the anestrous group. Compared with the anestrous group, a significant drop in mean LH content was seen by days 40-50 of pregnancy (894 vs 350 mug, P less than 0.