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Biomedical subjects

G E Lamming

Publications and source records attributed to G E Lamming.

At least 19 recordsLinked to original sources
PubMed1992

Plasma and luteal concentrations of oxytocin in cyclic and early-pregnant cattle.

Concentrations of oxytocin were measured in corpora lutea obtained from heifers throughout the oestrous cycle and first 30 days of pregnancy. Values were low during the first 3 days of the cycle (less than 250 ng/g tissue), increasing to 1312 ng/g by Day 4. Values then further increased up to a maximum of 2344 ng/g on Day 12. Concentrations were similar in cyclic and pregnant animals throughout the midluteal phase and were maintained at approximately 1500 ng/g until the 18th (cyclic cows) or 19th (pregnant cows) day after oestrus, when they were again low. Values subsequently remained less than 250 ng/g in pregnant cattle. Concentrations of oxytocin in jugular venous plasma of cyclic (n = 5) and pregnant (n = 4) cows were measured in samples collected every 15 min for 8 h on Days 14, 16, 18 and 19 after oestrus. There were no significant differences in mean concentrations (range: 2.5-4.7 pg/ml) or in the number, frequency or area under the curve of episodes between either cyclic and pregnant animals, or between days. Mean basal concentrations were higher on Day 16 than on Day 14 (P less than 0.05), values on Days 18 and 19 being intermediate. These findings suggest that the corpus luteum contains a finite amount of releasable oxytocin, which is exhausted by Day 18-19 after oestrus, whether or not pregnancy occurs, and that there is no further accumulation of oxytocin in the animal during early pregnancy. The contribution of luteal oxytocin to jugular venous concentrations appears to be less than in sheep, in which values in the jugular vein closely parallel those within the corpus luteum.

Animals↗
PubMed1992

Administration of recombinant bovine interferon-alpha I at the time of maternal recognition of pregnancy inhibits prostaglandin F2 alpha secretion and causes luteal maintenance in cyclic ewes.

The antiluteolytic protein, ovine trophoblast protein-1, which is secreted by sheep embryos at about the time of the maternal recognition of pregnancy, exhibits significant structural homology with alpha interferons. Experiments were conducted to examine the effects of intra-uterine and systemic administration of a recombinant bovine interferon-alpha I (rboIFN-alpha I) upon the interoestrus interval, endometrial oxytocin receptor concentrations and secretion of prostaglandin (PG) F2 alpha in cyclic ewes. In Expt 1, each ewe had a cannula placed in the tip of a uterine horn ipsilateral to a corpus luteum, 7 days after an induced oestrus. From day 9 after oestrus until day 19, ewes received either 200 (n = 4), 667 (n = 5) or 2000 (n = 9) micrograms/24 h of rboIFN-alpha I, meclofenamic acid (n = 4) or vehicle (n = 11). Other ewes received 2000 micrograms rboIFN-alpha I/24 h (n = 5) between days 12 and 15 only. All ewes were killed on day 19. Mean luteal phase, as determined by daily plasma progesterone measurements, was significantly longer (P less than 0.01) and mean concentrations of 13,14-dihydro-15-keto PGF 2 alpha (PGFM) in plasma were lower (P less than 0.05) in ewes receiving 667 or 2000 micrograms rboIFN-alpha I between days 9 and 19, or 2000 micrograms between days 12 and 15, than in animals from other treatment or control groups. A similar protocol was used in Expt 2, in which further ewes received either 2000 micrograms rboIFN-alpha I/24 h (n = 5) or vehicle (n = 5) by bolus infusions twice a day into one uterine horn. Mean luteal phase was significantly (P less than 0.05) longer in treated than in control animals, but differences in PGFM concentrations were not significant. In Expt 3, after a synchronized oestrus, ewes received either 2.5 mg rboIFN-alpha I by i.m. injection twice a day between days 12 and 15 (n = 10), 2.5 mg rboIFN-alpha I by i.m. injection twice a day between days 9 and 15 (n = 11), i.m. injection of vehicle alone twice a day (n = 20), or continual intra-uterine infusion of 2 mg rboIFN-alpha I/day between days 12 and 15 (n = 7). The mean luteal phase of ewes receiving rboIFN-alpha I by intrauterine infusion or i.m. injection between days 9 and 15 was significantly longer than for animals from the other two groups (P less than 0.05).(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗
PubMed1992

Role of the oxytocin receptor in the choice between cyclicity and gestation in ruminants.

The endometrial oxytocin receptor occupies a central point in the choice between luteolysis and pregnancy in ruminants. Receptor expression determines the time at which luteolysis occurs in nonpregnant animals, and thereby determines the duration of the oestrous cycle. Inhibition of receptor expression by trophoblast interferon (IFN) blocks the process of luteal regression and leads to continued progesterone secretion and the successful growth of the conceptus. Trophoblast IFN is probably required from the time of normal luteolysis during the cycle until luteal oxytocin concentrations of pregnancy decline, thus removing the stimulatory mechanism for the release of the episodes of prostaglandin F2 alpha required for luteolysis.

Animals↗
PubMed1992

Experimental and practical approaches to the establishment and maintenance of pregnancy.

A series of three large field trials was carried out to assess the effect of buserelin on fertility in dairy cows. In the first, 10 micrograms buserelin was injected on the day of insemination. There were no significant effects on fertility parameters compared to untreated controls. In the second trial cows were injected on day 12 after insemination. Mean pregnancy rates to first insemination were 53.4 and 65.4% for control and treated cows respectively (P < 0.01). Mean pregnancy rates to repeat inseminations were 52.9 and 59.4% for control and treated cows (NS). Mean calving to conception intervals were 91.4 and 85.3 days (P < 0.01) and the incidence of barren cows was 10.2 and 5.3% (NS). Overall the economic benefit of buserelin injection on day 12 was calculated to be 27.43 pounds per cow treated excluding the cost of the treatment. In trial 3 cows were injected with buserelin either on day 8 or 10 after insemination. There were no significant effects on fertility parameters compared to untreated control cows. In a fourth trial ewes were injected with 4 micrograms buserelin on day 12 after service. There were indications that both pregnancy rate and lambing percentage could be increased by buserelin treatment. Daily blood samples were collected from 5 dairy cows during a control cycle and a cycle in which 10 microgram buserelin was injected on days 11 and 13. Cycle length was unaffected by treatment and the concentration and pattern of progesterone secretion did not differ between control and treated cycles. Plasma oestradiol concentrations were similar in the control and treated cycles before day 11. However from day 12 to 16, equivalent to the time of maternal recognition of pregnancy, the mean concentration of oestradiol was significantly reduced in the treated cycle. As oestradiol stimulates both the development of uterine oxytocin receptors and the secretion of PGF2 alpha we suggest that any improvement in pregnancy rate after buserelin is due to a weakened luteolytic mechanism, resulting from a lower plasma oestradiol concentration.

Animals↗
PubMed1991

Uterine oxytocin receptors in cyclic and pregnant cows.

Binding of [3H]oxytocin to uterine subcellular preparations ('oxytocin receptor concentrations') was measured in uterine tissue of heifers and multiparous dairy cows at various stages of the oestrous cycle and during early pregnancy. A method for the assay of ovine uterine oxytocin receptors was optimized for use on bovine tissue. Oxytocin receptor concentrations were increased in cyclic animals around the period of luteolysis and oestrus, rising on Day 15 in endometrium and on Day 17 in myometrium while pregnant animals showed no comparable rise. Receptor concentrations then declined on Day 3 after oestrus in myometrium and on Day 5 in endometrium. Some cyclic animals did not show the expected rise in receptors in the late luteal phase; these animals had abnormally high progesterone concentrations for this stage of the cycle. In animals slaughtered on Day 18 after oestrus and/or insemination which had low oxytocin receptor levels, plasma progesterone concentrations were consistently high; while all animals showing the late luteal phase elevation in receptor values had low progesterone concentrations. Oxytocin receptor and progesterone concentrations were negatively correlated (P less than 0.05). These data support the hypothesis that oxytocin receptor level is a key factor in the process of luteolysis in cattle and that in pregnancy there is suppression of uterine oxytocin receptor at the expected time of luteolysis. We suggest that uterine oxytocin receptor levels are partly controlled by circulating steroid hormones and are suppressed during early pregnancy.

Animals↗
PubMed1991

A low molecular weight endometrial secretory protein which is increased by ovine trophoblast protein-1 is a beta 2-microglobulin-like protein.

Ovine trophoblast protein-1 (oTP-1), stimulates the secretion of several proteins in explant culture of day-12 cyclic ovine endometrium. We partially purified and identified one of these proteins, an 11,000 Mr, pI approx. 6 protein by N-terminal amino acid sequencing and immunoprecipitation using antibody to human beta 2-microglobulin. The protein was purified from cultures of endometrium collected from day-16 pregnant ewes. The N-terminal amino acid sequence was 40-55% homologous to beta 2-microglobulin from a variety of species. Antibody to human beta 2-microglobulin immunoprecipitated the protein and another protein of similar molecular weight but more acidic pI. Using immunoprecipitation of radiolabelled proteins from culture, we demonstrated that oTP-1 increased production of this protein by 40% (P less than 0.05). We conclude that oTP-1 increases the secretion of a beta 2-microglobulin-like protein from day-12 non-pregnant endometrium in culture.

Amino Acid Sequence↗
PubMed1991

Ovine conceptus secretory proteins and bovine recombinant interferon alpha (1)-1 decrease endometrial oxytocin receptor concentrations in cyclic and progesterone-treated ovariectomized ewes.

A series of experiements was performed to determine whether proteins produced by the sheep conceptus (oCSP) during the time of maternal recognition of pregnancy or bovine recombinant interferon alpha 1-1 (brIFN) decrease oxytocin receptor concentrations in the endometrium of cyclic or ovariectomized progesterone-treated ewes. In experiment 1, cyclic ewes received intrauterine infusions of serum proteins (oSP), oCSP or brIFN on days 12, 13 and 14 of the oestrous cycle. Ewes then received an oxytocin challenge (1 microgram in 0.9% NaCl), and blood samples were taken just before and every 10 min for 1 h after the challenge; these were measured for 13,14-dihydro-15-ketoprostaglandin F 2 alpha (PGFM), the stable metabolite of prostaglandin F 2 alpha. Endometrial oxytocin receptor concentrations were then measured. The oCSP and brIFN treatments suppressed both endometrial oxytocin receptor concentrations and oxytocin-induced increases in PGFM concentrations. In experiment 2, ewes were ovariectomized and then pretreated with a fluorogestone acetate-releasing intravaginal device for 10 days followed by oestradiol (25 micrograms i.m. twice daily for 2 days). Ewes were then treated with progesterone (10 mg i.m. twice daily for 12 days). Ewes received intrauterine infusions of oSP, oCSP and brIFN on days 10, 11 and 12 of progesterone treatment. On the day after the last progesterone treatment, ewes were challenged with oxytocin and blood samples collected to measure PGFM. Endometrial oxytocin receptors were also measured. Treatment with oCSP, but not brIFN, suppressed endometrial concentrations of oxytocin receptor, and neither oCSP nor brIFN altered oxytocin-induced increases in PGFM concentrations.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1991

Molecular biology of trophoblast interferons and studies of their effects in vivo.

Southern blotting of bovine genomic DNA indicated the presence of at least 3 bovine tIFN genes. The full DNA sequence of one of these genes, thought to be expressed in trophoblast, has been determined, including 193 bp of 5' non-coding region. The inferred amino acid sequence of bovine tIFN is more similar to ovine tIFN (80%) than to bovine IFN-alpha II (70%). The 5' flanking sequence has some similarity with bovine IFN-alpha II, and may contain a viral response element. A recombinant bovine alpha I interferon (Ciba Geigy; brIFN), resembling tIFN, extended oestrous cycle length in sheep when administered by intrauterine infusion over the period, Days 12-15 after oestrus, when maternal recognition of pregnancy occurs. Intramuscular injection was only effective at the doses used if given over a longer period (i.e. Days 9-15). Our experiments indicate that both tIFN and brIFN inhibit luteolysis by preventing a rise in endometrial oxytocin receptor concentrations, and suggest that tIFN achieves this by extending the time for which progesterone suppresses oxytocin receptor development. Further studies are required to confirm this hypothesis and to elucidate the interaction of the effects of progesterone and tIFN in endometrial cells.

Amino Acid Sequence↗
PubMed1990

Luteal inadequacy during the early luteal phase of subfertile cows.

A study was made of early luteal function (up to Day 6) in cyclic and pregnant heifers and also in older, subfertile cows. There were no differences in vivo or in vitro between cyclic and pregnant heifers, indicating no luteotrophic effect of the embryo at this stage, but the increase in postovulatory peripheral progesterone concentrations was delayed (P less than 0.01) and occurred more slowly (P less than 0.001) in the subfertile cows than in the heifers. The corpora lutea of the subfertile cows were heavier (P less than 0.001) than those of the heifers on Day 6. Basal progesterone production by dispersed luteal cells was similar between heifers and subfertile cows, but there was a difference (P less than 0.001) in the pattern of response to exogenous LH and PGE-2. Cells from subfertile cows were less sensitive to the stimulatory effects of PGE-2 and although LH increased (P less than 0.001) progesterone production by all cells, this stimulation by a low dose of LH was inhibited by PGE-2 in luteal cells from subfertile cows. This effect did not occur in the luteal cells from heifers. These results indicate the possibility that luteal inadequacy, due to a diminished response to circulating luteotrophic hormones, may contribute to embryo mortality in subfertile cows.

Animals↗
PubMed1990

Prostaglandin E-2 as a potential luteotrophic agent during early pregnancy in cattle.

Heifers slaughtered on Day 18/19 of pregnancy had significantly higher (P less than 0.001) concentrations of PGE-2 (measured as its methyl oxime) in uterine flushings than did animals slaughtered on Days 6 or 12 of pregnancy, or on Days 6 or 12 of the oestrous cycle. In addition, concentrations were higher in the uterine horn ipsilateral to the corpus lueum on Days 12 (P less than 0.05) and 18/19 (P less than 0.01) than in the contralateral horn. Incubation of dispersed luteal cells for 3 h with LH (0.1 or 100 ng/ml) and/or PGE-2 (0.01-1000 ng/ml) in vitro showed no differences in basal progesterone production or in the responses to exogenous hormones between pregnant and non-pregnant cattle. However, low doses of PGE-2 (0.01-10 ng/ml) inhibited the stimulation of progesterone secretion by the lower dose of LH. These findings indicate that although PGE-2 can stimulate progesterone synthesis by luteal cells it may also have inhibitory effects, and therefore its role in pregnancy requires further definition.

Animals↗
PubMed1990

Interrelationships between progesterone, 13,14-dihydro-15-keto PGF-2 alpha (PGFM) and LH in cyclic and early pregnant cows.

Plasma progesterone and LH secretion patterns were examined in 18 mature dairy cows during the oestrous cycle and after insemination. Blood samples were collected every 15 min for 8 h per day on Days 3, 5, 6, 7, 8, 9, 10, 12, 14, 16, 17, 18, 19, 20 and 21 of the oestrous cycle, then, in the same cows, at the same times during early pregnancy. PGF-2 alpha secretion rates (as determined by plasma PGFM concentrations) were also monitored on Days 14, 16 and the day of, or equivalent to, luteal regression. Mean daily plasma progesterone concentrations were similar until Day 16 in cyclic and pregnant cows, after which values in non-pregnant animals declined. Regression analysis indicated that progesterone concentrations were best described by a quadratic expression with fitted maximum values on Day 13 in non-pregnant animals but values increased linearly over the whole period to Day 21 in pregnant cows. The frequency, amplitude and area under the curve of LH episodes showed no significant differences between cyclic and pregnant animals. In pregnant cows, the amplitude and area under the curve of progesterone episodes increased linearly between Days 8 and 21, although no such increase occurred in cyclic cows. Low-level PGFM episodes were present in cyclic and pregnant cows on Days 14 and 16 after oestrus, and high amplitude episodes occurred in non-pregnant cows during luteal regression. Pregnant cows showed a significant depression of the amplitude, but not the frequency of episodes at the expected time of luteal regression. These results confirm that the corpus luteum of pregnancy secretes an increasing amount of progesterone per se and per unit of LH until at least Day 21 after mating. They further suggest that the corpus luteum of the cyclic cow may experience small episodes of PGF-2 alpha and be subjected to initial degenerative changes by Day 14 after oestrus, some time before the onset of definitive luteolysis.

Animals↗
PubMed1990

Comparison of oxytocin/prostaglandin F-2 alpha interrelationships in cyclic and pregnant cows.

The secretion of prostaglandin (PG) F-2 alpha in response to intravenous injection of 100 i.u. oxytocin on Day 18 after oestrus was determined by measuring jugular venous concentrations of 13,14-dihydro-15-keto PGF-2 alpha (PGFM) in 7 pregnant, 6 cyclic and 2 inseminated non-pregnant heifers. Two other heifers received i.v. saline (controls). The immediate responses of pregnant heifers were smaller than in non-pregnant animals (P less than 0.05), as were baseline concentrations in the post-response period (P less than 0.05). Endometrial oxytocin receptor concentrations were higher in nonpregnant than pregnant heifers (P less than 0.05), but PGFM response to oxytocin challenge was not correlated with oxytocin receptor concentration. Oxytocin receptor concentrations on Day 18 were positively correlated with those of plasma oestradiol on Day 17 (P less than 0.01) and inversely with plasma progesterone concentrations on Day 18 (P less than 0.01). These findings confirm that PGF-2 alpha secretion in response to oxytocin challenge is attenuated in pregnant animals on the 18th day after oestrus and that, while the prevailing steroid environment is of importance in inducing oxytocin receptor activity, the secretion of PGF-2 alpha is not subsequently limited by oxytocin receptor numbers. The quantities of PGE-2, PGFM and PGF-2 alpha recovered in uterine flushings taken from heifers on Day 18 were greater in pregnant than other animals (P less than 0.01, P less than 0.05, P less than 0.001, respectively). Intrauterine concentrations of PGF-2 alpha and PGFM were not correlated with the plasma PGFM responses.

Animals↗
PubMed1990

Control of endometrial oxytocin receptor and uterine response to oxytocin by progesterone and oestradiol in the ewe.

The effects of administration of progesterone and oestradiol on ovine endometrial oxytocin receptor concentrations and plasma concentrations of 13,14-dihydro-15-keto prostaglandin F-2 alpha (PGFM) after oxytocin treatment were determined in ovariectomized ewes. Ewes received progestagen pre-treatment, progesterone and/or oestradiol in 11 different treatment schedules. Progestagen pre-treatment decreased oxytocin receptor concentrations in endometrium from ewes treated subsequently with either progesterone for 5 days or progesterone for 5 days plus oestradiol on Days 4 and 5 of progesterone treatment. Oestradiol increased endometrial oxytocin receptor concentrations when administered on Days 4 and 5 of 5 days progesterone treatment. Progestagen pre-treatment followed by progesterone treatment for 12 days caused a large increase in oxytocin receptors and no further increase occurred when ewes were given oestradiol on Days 11 and 12, or when progesterone was withdrawn on Days 11 and 12, or these two treatments were combined. Oxytocin administration caused an increase in plasma PGFM concentrations in ewes which did not receive progestagen pre-treatment, and subsequently received progesterone treatment for 5 days and oestradiol treatment on Days 4 and 5 of progesterone treatment. Similarly treated ewes which received progestagen pre-treatment did not respond to oxytocin. Oxytocin administration also increased plasma PGFM concentrations in ewes which received progestagen pre-treatment followed by progesterone treatment for 12 days, progesterone treatment for 12 days plus oestradiol on Day 11 and 12 of progesterone treatment, progesterone withdrawal on Day 11 and 12, or progesterone withdrawal and oestradiol treatment combined. The results indicate that (1) progesterone pre-treatment affects oxytocin receptor concentrations in the endometrium and uterine responsiveness to oxytocin and (2) progesterone treatment alone for 12 days after a treatment which mimics a previous luteal phase and oestrus is sufficient to induce oxytocin receptors and increase oxytocin-induced PGF release. These results emphasize the importance of progesterone and provide information which can be used to form an hypothesis for control of luteolysis and oestrous cycle length in the ewe.

Animals↗
PubMed1989

Failure of melatonin to influence endogenous opioid effects on LH secretion in the anoestrous ewe.

In May mature seasonally anoestrous ewes were implanted with melatonin which advanced the onset of cycles by about 1 month. The LH response to an opioid antagonist, WIN-3, was determined 5, 15, 25 and 60 days after melatonin implantation, by intravenous administration of WIN-3 (12.5 mg/dose) 4 times at 15-min intervals during both the 1st and the 5th hour of an 8-h treatment period. There was no effect of WIN-3 at 5, 15 and 25 days after melatonin implantation. At 60 days LH concentration and pulse frequency were significantly increased (P less than 0.05 and less than 0.01 respectively) in response to WIN-3 treatment, but only in those animals which had begun reproductive cycles, an effect known to be mediated by the presence of progesterone. We were therefore unable to find evidence to support the hypothesis that the influence of melatonin in advancing the breeding season may be via an opioidergic pathway.

Anestrus↗
PubMed1989

Oxytocin infusion from day 10 after oestrus extends the luteal phase in non-pregnant cattle.

Oestrus was synchronized in 8 cyclic heifers by progesterone treatment (PRID), after which the animals were monitored for one control cycle to measure the inter-oestrous interval. Osmotic minipumps containing saline (controls, N = 3) or oxytocin (N = 5) were implanted subcutaneously on Day 10 of the second cycle, and removed 12 days later. Jugular venous blood samples were collected daily for measurement of progesterone, and every 2 days for oxytocin. In addition, blood samples were taken every 10 min from 1 h before to 3 h after minipump insertion for measurement of plasma 15-keto-13,14-dihydroprostaglandin-F-2 alpha (PGFM) and every 30 min over the same period for measurement of progesterone and oxytocin. The lengths of the first untreated cycle in both groups of heifers were 20.2 +/- 0.56 (mean +/- s.e.m.) days compared with 25.4 +/- 0.81 days after oxytocin treatment (P less than 0.001). Oxytocin plasma concentrations in treated animals rose from less than 10 pg/ml to 70-500 pg/ml by 2 h after the start of oxytocin infusion and remained elevated until treatment was withdrawn. There was no increase in PGFM concentrations immediately after minipump insertion. Plasma progesterone concentrations were similar in treated and control animals but remained at mid-luteal levels for an average of 5 days longer in treated heifers. It is concluded that continuous administration of oxytocin can extend the luteal life-span in cattle.

Animals↗
PubMed1989

Sheep antiluteolytic interferon: cDNA sequence and analysis of mRNA levels.

A cloned cDNA has been isolated by probing a sheep blastocyst cDNA library using a synthetic oligonucleotide representing the N-terminal amino acid sequence of the antiluteolytic protein, ovine trophoblast protein-1. Sequence analysis of the cDNA confirms the 70% homology between the antiluteolysin and the interferon-alpha family of proteins; however, the sequence reported here differs at several points from previously reported amino acid and cDNA sequences for the antiluteolysin. In-vitro translation of day-16 poly(A)+ RNA indicated that antiluteolysin mRNA is a major constituent of total mRNA at this stage of blastocyst development, and Northern blotting confirmed that antiluteolysin mRNA production occurred between days 13 and 22 after oestrus. This is consistent with the stage at which embryonic extracts are antiluteolytic on administration in vivo. These and other data confirm that the ovine trophoblast antiluteolysin is an interferon, and suggest that at least five isoforms of this protein may exist.

Amino Acid Sequence↗
PubMed1989

Quantification of opioid-binding sites in the ewe hypothalamus.

Opioid-binding sites were quantified in the ewe hypothalamus using [3H]diprenorphine ([3H]DIP) as the radioligand. [3H]DIP binding to hypothalamic membrane preparations was stereospecific, saturable with respect to [3H]DIP concentration, and linear with hypothalamic membrane protein content. Scatchard analysis revealed a single class of binding sites. There were no significant differences in binding site concentration or binding affinity in hypothalami from intact ewes during the breeding and non-breeding seasons, or from long-term ovariectomized ewes with and without oestradiol treatment during the breeding season. Thus, whilst ovarian steroid hormones are known to modify LH responses to opioids and their antagonists in the ewe in vivo, they do not appear to do this by modulating the numbers of hypothalamic opioid-binding sites.

Animals↗