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Biomedical subjects

J A Downing

Publications and source records attributed to J A Downing.

30 records · Page 2Linked to original sources

Changes in progesterone secretion following treatment with transforming growth factor alpha (TGF-alpha) during the follicular phase of the sheep oestrous cycle.

The effects of transforming growth factor alpha (TGF-alpha) on ovarian steroid secretion were investigated. Three crossbred ewes synchronized for oestrus with ovarian autotransplants were infused with TGF-alpha (30 micrograms in 12 h) via the ovarian artery for 12 h before withdrawal of progestagen pessary. Three ewes were used as controls. Jugular and ovarian venous blood samples were taken at intervals of 10 min at two stages during the follicular phase (21-27 h and 38-42 h after pessary withdrawal) and every 2 h from 44 to 86 h. Plasma LH and FSH concentrations, and ovarian secretion rates of inhibin, androstenedione, oestradiol and progesterone were determined using radioimmunoassays. LH pulse amplitude increased in ewes treated with TGF-alpha in the early follicular phase (0.92 +/- 0.25 micrograms l-1 in controls versus 3.10 +/- 0.35 micrograms l-1 in TGF-alpha treated ewes; P < 0.05) and remained high in the late follicular phase. Plasma FSH concentrations were high during the follicular phase in ewes treated with TGF-alpha (P < 0.05). The infusion of TGF-alpha had no significant effect on the ovarian rate of secretion of androstenedione and, although the secretion rates of oestradiol and inhibin were consistently lower in TGF-alpha-infused ewes, the differences were not significant. The ratio of secretion of androstenedione to oestradiol was greater during the follicular phase in TGF-alpha-treated ewes (P < 0.05), suggesting that the efficiency of aromatization had been impaired.(ABSTRACT TRUNCATED AT 250 WORDS)

Androstenedione↗

A model for follicle selection and the determination of ovulation rate in the ewe.

A model for folliculogenesis is proposed that is based as far as possible on a knowledge of physiological, rather than anatomical, changes taking place during follicle development. The model is therefore functional, rather than descriptive, and consists of five classes of follicles that have been defined by their dependency and sensitivity to gonadotrophins. These classes are: primordial, committed, gonadotrophin-responsive, gonadotrophin-dependent and ovulatory. The model is an attempt to encourage discussion and to promote the integration of morphological models of folliculogenesis with recent advances in the molecular endocrinology of the ovarian follicle. Two hypotheses for the mechanisms that determine ovulation rate are developed in light of the model. In the first, multiple ovulation results when the viability of gonadotropin-dependent follicles is enhanced. In the second, multiple ovulation is caused by increasing the number of gonadotrophin-responsive follicles available for further development; this results from the increasing rate of folliculogenesis and the throughput of follicles. The final section of this paper examines how these two hypothetical mechanisms, which are not mutually exclusive, appear to account for most of the known genetical and environmental effects on ovulation rate of sheep. In particular, the effects of nutrition, genotype, exogenous gonadotrophins, immunity to both oestrogens and androgens, and immunity to inhibin are discussed.

Animal Nutritional Physiological Phenomena↗

Epidermal growth factor acts directly on the sheep ovary in vivo to inhibit oestradiol-17 beta and inhibin secretion and enhance progesterone secretion.

Epidermal growth factor (EGF) is a potential intra-ovarian modulator of gonadotroph action on differentiated follicular cells. Specific binding sites have been identified in the ovary and functional differentiation in cultured granulosa cells can be modulated by treatment with EGF. The aim of this study was to determine if EGF was capable of altering ovarian function in vivo during the follicular phase of the sheep oestrous cycle. Fourteen cross-bred ewes with ovarian autotransplants were treated with progestagen pessaries for 12 days. Three ewes were infused with murine EGF (mEGF) via the jugular vein (75 micrograms/kg bodyweight per 12 h) during the 12 h preceding progestagen pessary withdrawal, and received an injection of a prostaglandin analogue at 0 h to induce luteolysis. Over the same time-period, two doses of EGF were administered to other groups of ewes by infusion into the ovarian artery (low: 6 micrograms/12 h, n = 3 and high: 60 micrograms/12 h, n = 3). The remaining five ewes were not infused with EGF (controls). Jugular and ovarian venous blood samples were taken at 10-min intervals at two stages during the follicular phase (21-27 h and 38-42 h after pessary withdrawal) and every 2 h from 44 to 76 or 86 h. mEGF, LH, FSH, inhibin, androstenedione, oestradiol-17 beta and progesterone concentrations in plasma were determined using radioimmunoassays. The secretion rates of androstenedione, oestradiol, progesterone and inhibin by the ovary were calculated. EGF acted directly on the ovary in a dose-dependent manner. Oestradiol secretion was inhibited following treatment with EGF but androstenedione secretion was unaffected. EGF appears therefore to act within the granulosa cells to inhibit aromatization. Inhibin secretion was also suppressed by treatment with EGF, though it was not possible to determine if this was caused by a direct or indirect action of EGF on granulosa cells. The rate of progesterone secretion increased in ewes receiving systemic (i.e. via the jugular vein) and high-dose intra-arterial infusions of EGF, even though a preovulatory LH surge was not observed in these animals during the entire experimental period. Concomitant increases in both LH and FSH secretion were associated with these effects of EGF on ovarian function. In conclusion, EGF appears to act directly on the granulosa cells of the follicle to inhibit aromatization and also to inhibit inhibin production. The low levels of oestradiol and inhibin in the presence of high levels of gonadotrophin indicate that atresia may have been induced in medium to large antral follicles.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗

Increased ovulation rate in androstenedione-immune ewes is not due to elevated plasma concentrations of FSH.

Two experiments were undertaken to determine the hormonal response of Merino ewes to immunization against androstenedione (Fecundin). In Exp. 1 peripheral concentrations of LH, FSH and progesterone were monitored in spontaneously cycling ewes (20 immunized and 21 controls). In Exp. 2 (10 immunized and 10 controls) the same hormones were measured in ewes before and after prostaglandin (PG)-induced luteolysis and, in addition, the pattern of pulsatile LH secretion was determined during the luteal (PG + 12 days), early follicular (PG + 24 h) and late follicular (PG + 40 h) phase of the oestrous cycle. Ovulation rates were measured in both experiments. The results of these experiments indicate that androstenedione-immune animals have elevated ovulation rates (0.6-0.7 greater than control animals; P less than 0.05) associated with elevated plasma concentrations of LH and progesterone. The magnitude of the increase in plasma progesterone was correlated with androstenedione antibody titre (r = 0.6, P less than 0.001). LH pulse frequency of androstenedione-immune ewes tended to be higher at all stages of the oestrous cycle, but this difference was only significant (P less than 0.05) during the luteal phase. Mean plasma concentrations of FSH did not differ significantly between immunized and control ewes at any stage of the cycle. Analysis of periodic fluctuations in FSH during the luteal phase revealed that androstenedione-immune animals had a similar number of fluctuations of a similar amplitude to those of control animals, but the nadir of these fluctuations was lower (P less than 0.05) in immunized animals. A significant (P less than 0.05) negative correlation existed between androstenedione antibody titre and the interval between FSH peaks (r = -0.49) and androstenedione antibody titre and FSH nadir concentrations (r = -0.46). It is concluded that plasma FSH concentrations are not a determinant of ovulation rate in androstenedione-immune ewes and that increased LH concentrations, or perturbation of normal intraovarian mechanisms, may be responsible for the increase in ovulation rate observed in ewes immunized against androstenedione.

Androstenedione↗

Steroid secretion rates and plasma binding activity in androstenedione-immune ewes with an autotransplanted ovary.

Mature Merino ewes in which the left ovary and its vascular pedicle had been autotransplanted to the neck were divided into control (N = 5) and immunized groups (N = 6). The immunized ewes were treated (2 ml s.c.) with Fecundin 1 and 4 weeks before the start of blood sampling. Ovarian and jugular venous blood was collected every 10 min at two stages of the follicular phase (21-27 h and 38-42 h after i.m. injection of 125 micrograms of a prostaglandin (PG) analogue) and during the mid-luteal phase (8 h at 15-min intervals). The ewes were monitored regularly for luteal function and preovulatory LH surges. Hormone concentrations and anti-androstenedione titres were assayed by RIA and ovarian secretion rates of oestradiol-17 beta, progesterone and androstenedione were determined. After the booster immunization, progesterone increased simultaneously with titre in immunized ewes, reaching 30 ng/ml at the time of PG injection when median titre was 1:10,000. All ewes responded to PG with LH surges 42-72 h later: 2 of the immunized ewes then had a second LH surge within 3-4 days at a time when peripheral progesterone values were 2-3 ng/ml. The frequency of steroid and LH pulses was greater in immunized ewes (P less than 0.05) during the luteal phase but not the follicular phase. The secretion rate of androstenedione was 6-10 times greater (19-37 ng/min; P less than 0.001) in immunized ewes at all sampling stages. Progesterone secretion rates were 3 times greater (16 micrograms/min; P less than 0.001) during the luteal phase in immunized ewes. The amplitude of oestradiol pulses was significantly reduced in immunized ewes (4.8 vs 2.1 ng/min at +24 h and 6.5 vs 2.8 ng/min at +40 h in control and immunized ewes, respectively: P less than 0.05) during the follicular phase. However, the mean secretion rate of oestradiol at each phase of the cycle was not significantly different between treatment groups. Analysis of bound and free steroid using polyethylene glycol showed that greater than 98% of peripheral and ovarian venous androstenedione and 86% of peripheral progesterone was bound in immunized ewes but there was no appreciable binding (less than 0.1%) in control ewes. Similarly, 50% of ovarian venous oestradiol was bound in immunized ewes compared to 15% in control ewes. We conclude that immunization against androstenedione increases the secretion rate of androstenedione and progesterone but not of oestradiol.(ABSTRACT TRUNCATED AT 400 WORDS)

Androstenedione↗

Seasonal modification of ovine pineal function. I. Interaction of gonadal steroids and sympathetic innervation on beta-adrenoceptors.

The results of a seasonal study of the regulation by steroids of ovine pineal beta-adrenoceptors are reported. The effects of ovariectomy, steroid implants and superior cervical ganglionectomy were investigated in Merino ewes. The results confirm earlier observations showing a seasonal variation in the sensitivity of the receptors to modification by gonadal steroids. A marked difference in receptor density and affinity between the follicular and luteal phases of the estrous cycle is shown for the first time, a difference which can be modified by appropriate steroid-related treatments. A season-dependent antagonism between the sympathetic innervation of the gland (presumably acting via the neurotransmitter noradrenaline) and gonadal steroids in the regulation of ovine pineal beta-adrenoceptor function is also shown.

Animals↗

Seasonal modification of ovine pineal function. 2. Steroidal effects on melatonin and prolactin profiles.

Pineal beta-adrenoceptor density and affinity in ewes are modified in a season-dependent manner by gonadal steroids and by the sympathetic innervation of the gland. The present study was undertaken to relate the steroidal effects on the receptors to post receptor endocrine events, and to investigate the influence of the sympathetic innervation of the pineal gland on these events. Plasma melatonin and prolactin profiles were determined during anestrus and during the normal breeding season in ewes subjected to sympathetic denervation of the pineal and/or a range of steroid-related treatments. Wherever valid comparisons could be drawn between effects of the treatments on beta-adrenoceptor variables and on circulating hormone levels, similar effects were noted. Further, ganglionectomy influenced hormone profiles similarly to estradiol under all conditions tested. It appears that gonadal steroids (estradiol) and the sympathetic neurotransmitter noradrenaline have opposing actions on prolactin levels, just as they have on pineal beta-adrenoceptor binding affinity. These findings suggest that steroid-mediated changes in receptor number and affinity are reflected in post receptor endocrine events. In addition, other factors (e.g. photoperiodic information transmitted via the sympathetic innervation) also play important roles in the regulation of the observed hormonal profiles. A steroid-mediated feedback regulation of pituitary prolactin release, partly direct and partly via pineal melatonin release, is suggested.

Anestrus↗

Secretion of bioactive inhibin by the ovary of the Booroola Merino ewe with or without a copy of the fecundity (F) gene.

The secretion rates of bioactive inhibin, oestradiol and progesterone were measured during the mid-luteal phase and at various times during the follicular phase of the cycle by a sensitive bioassay using sheep pituitary cells in culture in 12 Booroola ewes with and without copies of the Fecundity (F) gene in which the left ovary had been auto-transplanted to the neck. Inhibin secretion was high during the luteal phase and fell in the early follicular phase in all genotypes (P less than 0.01). In Booroola ewes with a F/- genotype, inhibin secretion then increased again, towards luteal rates, in the mid and late follicular phases. In Booroola ewes without a copy of the F gene (+/+) inhibin secretion remained low at all three sampling times in the follicular phase. The secretion rate of inhibin at 36 h (P less than 0.1) and 48 h (P less than 0.01) were significantly lower in ewes from the +/+ (no copy of the gene) ewes than in F/- (one copy of the gene) ewes. Oestradiol secretion was low during the luteal phase and increased steadily during the early (24 h) to a plateau in the mid (36 h; P less than 0.01) and late (48 h; P less than 0.05) follicular phase. Progesterone secretion was high during the luteal phase, and decreased to a very low rate by 24 h after prostaglandin (PG) treatment (P less than 0.001) and remained low. At 24 h after PG the concentration of FSH was significantly lower (P less than 0.01) than that during the luteal phase and remained suppressed until the onset of the LH surge.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Control of fertility and fecundity of sheep by means of hormonal manipulation.

The results of four experiments are presented in summary form. The data are considered in relationship to the improvement of the fecundity and fertility of the Australian ewe breeding flock. In the first, three commonly used methods of oestrous synchronization were examined and showed differences that are attributed to the different patterns of hormonal changes associated with the methods demonstrated. The second experiment looked at the use of active immunization against testosterone and concluded that this method can improve fecundity but not fertility. The third experiment, a group of five trials, studied the use of progestagen sponges and PMSG in anoestrous ewes as a means of inducing normal fertility. The extensive data produced in this experiment allowed the relationships between ovulation rate and fertility and between fertility and prolificacy (fecundity) to be examined. Fertility appeared greatest when the mean flock ovulation rate was about 2.5. At this ovulation rate prolificacy was also improved and a high proportion of twins were produced. We concluded that high fertility and low prolificacy (i.e. of 1.00) are an unlikely combination. In the final experiment the effect of post-mating hormonal supplementation on fertility was examined and a number of earlier reports were confirmed by showing that fertility can be improved with supplementary progesterone between days 10 and 25 post-mating. The effect appears to be modified by hormonal and nutritional factors.

Animals↗

Soil phosphorus, management practices, and their relationship to phosphorus delivery in the Iowa Clear Lake agricultural watershed.

Clear Lake is on Iowa's list of impaired water bodies because of high P concentration. This study assessed soil-test phosphorus (STP), management practices, and P loads from its agricultural watershed. Management practice histories and STP for eight basins were surveyed in 1999. Soil samples (15-cm depth) were analyzed for STP with agronomic [Bray P1 (BP), Olsen (OP), Mehlich 3 (M3P) and environmental [iron oxide-impregnated paper (FeP) and water extraction (WP)] tests. Total phosphorus (TP) concentrations in water discharge from five basins were measured during two years, and TP loads were measured for two basins. The agronomic P tests showed that 46 to 83% (depending on the test) of the area tested above optimum for crops. Correlations among tests were high for OP, M3P, and FeP (r > 0.96) and lower for BP and WP (r = 0.88-0.93). Moldboard- and chisel-plow tillage predominated (82% of the area). Applied P (mainly fertilizer) averaged 15 kg P ha(-1) yr(-1), and 40% of the high-testing area (M3P test) was being fertilized. The mean annual water TP concentration across five basins was 275 to 474 microg L(-1). The two-year mean TP loads for the two gauged basins were 1504 and 1510 g P ha(-1) yr(-1). Water TP concentration increased linearly with increasing STP. Relationships were stronger for M3P and FeP (R2 = 0.96-0.97 for annual means and 0.77-0.79 for storm-flow events) than for BP or WP (R2 = 0.88-0.91 and 0.59-0.69, respectively). Improving P and soil conservation practices in high-testing areas could reduce P loads to the lake.

Conservation of Natural Resources↗