Biomedical subjects
D G Porter
Publications and source records attributed to D G Porter.
Relationship between pre-partum relaxin concentrations and farrowing intervals in the pig.
Gilts were treated on Day 112 of gestation with saline or a prostaglandin (PG) F-2 alpha analogue. In control gilts there was a rise in the relaxin concentration from 48 h before the onset of delivery, peaking between 12 and 28 h pre partum followed by a steep fall. The relaxin concentrations at each 1-h time interval were analysed in relation to the farrowing interval for each gilt using correlation analysis. There was a significant (at least P less than 0.05) positive correlation between the relaxin concentration and the farrowing interval at every time period from 14 to 2 h before delivery. In contrast there was little relationship between concentrations of progesterone, oestrone and oestradiol-17 beta and farrowing intervals. The gilts treated with PGF-2 alpha analogues had steroid profiles indistinguishable from those in controls but differing relaxin secretion patterns. Relaxin concentrations peaked at 1-2 h after PGF-2 alpha injection and this was followed by a second smaller increase closer to the time of delivery in 7 out of 12 gilts. The 'two-peak' gilts had significantly higher relaxin concentrations at farrowing and took significantly longer to farrow than did the 'one-peak' gilts (P less than 0.005). These results suggest that high relaxin concentrations during the last 14 h before the onset of parturition are associated with increased farrowing times, but are not associated with any increase in neonatal mortality.
Plasma steroid, relaxin and dihydro-keto-prostaglandin F-2 alpha changes in the minipig in relation to myometrial electrical and mechanical activity in the pre-partum period.
Recordings of electromyographic and mechanical activities of the uterus of intact and ovariectomized minipigs during the last days of pregnancy, combined with a frequent blood sampling regimen, indicated that the evolution of electrical activity was dependent upon the clearance both of progesterone and relaxin from the plasma. Increases in dihydro-keto-prostaglandin-F-2 alpha (DHKF-2 alpha) occurred only after the initial decline in progesterone, and were positively correlated with increases in electrical activity only after circulating relaxin titres had fallen. These results demonstrate that relaxin provides for the inhibition of the myometrium during the period when progesterone titres decline. The temporal relationship of the decline of progesterone and the rise in DHKF-2 alpha suggests that uterine PGF-2 alpha may not initiate luteolysis in the sow at term.
Identification of relaxin in the placenta of the ewe.
Sheep placentomes were collected at the abattoir and the stage of gestation was estimated from the crown-rump length and appearance of the fetus. Samples were extracted and either freeze dried (crude extracts) or fractionated on Sephadex G-50 and CM-cellulose. Relaxin immunoreactivity (RXN-IR) was detected in all samples by a pig relaxin RIA and diluted in parallel with the standard curve. Two patterns of RXN-IR were seen after Sephadex G50 purification: (a) a single main peak of RXN-IR eluting at a position similar to pig relaxin; or (b) a 3-peak pattern with additional higher (void volume) and lower (approximately 1000) molecular weight peaks. These peaks were all found with 4 different and specific antisera. The 6000 molecular weight peak eluted at a similar position to pig relaxin on CM cellulose and inhibited electrically stimulated rat uterine contractions in vitro. The amount of relaxin measured in crude extracts of placentomes from different ewes was very variable. Most samples were within the range 0.05-11.2 ng/g wet weight of tissue (3.0 +/- 0.45 (s.e.m.), n = 44) but a few contained much higher concentrations (25.5-61.4 ng/g, n = 3). There was no obvious variation in concentration with stage of pregnancy (20 days to term). Samples of intercotyledonary endometrium, allantochorion and whole ovaries from pregnant ewes were also extracted. All contained low concentrations of RXN-IR (0.6 +/- 0.13 ng/g, n = 4; 0.6 +/- 0.29 ng/g, n = 3; 1.0 +/- 0.66 ng/g, n = 7, respectively). We conclude that relaxin-like peptides are present in the pregnant ewe and that, as the placentomes are the largest component by weight, they represent the major source.
Identification of relaxin immunoreactivity in human follicular fluid.
Relaxin immunoreactivity has been found in samples of human follicular fluid collected from artificially stimulated pre-ovulatory follicles. The crude extract caused a reduction in the height of the contractions in a rat uterine strip bioassay. The reactive material eluted from Sephadex G50 in two major peaks. The first contained approximately 60% of the immunoreactivity and had an elution position corresponding to that of porcine relaxin, indicating a mol. wt of approximately 6000 daltons. The second peak was of a lower mol. wt, but its exact size and significance are unknown. A possible role for relaxin in the process of follicular rupture is suggested.
Relaxin and progesterone are myometrial inhibitors in the ovariectomized non-pregnant mini-pig.
Intravenous bolus injections of pig relaxin which produced short-lived peaks of the hormone equivalent in concentration to those observed at term promptly rendered the uterus almost totally quiescent and the inhibition persisted for about 2.5 h. During this time the uterus remained responsive to oxytocin. The main effect of relaxin was to reduce the frequency of intrauterine pressure cycles rather than the amplitude. In contrast progesterone, which also inhibited myometrial activity, took between 6 and 24 h to exert its maximum effect by reducing both amplitude and frequency of IUP cycles and it also abolished the responsiveness of the myometrium to oxytocin. Its actions were reversible but recovery took between 54 and 140 h. Oestradiol benzoate had no significant effect on myometrial activity in 21 out of 26 treatments. At 24 h after the 5 remaining treatments, however, myometrial activity was virtually zero. No evidence was obtained of a biphasic effect of oestradiol on myometrial activity as reported for the rat and ewe. This work demonstrates that purified pig relaxin is an active myometrial inhibitor in the oestrogen-treated ovariectomized non-pregnant pig in vivo.
A combined radioimmunoassay and immunocytochemical study of ovarian oxytocin production during the periovulatory period in the ewe.
Corpora lutea and follicles were taken from the ovaries of 12 ewes at intervals from the start of luteolysis until 3 days after ovulation. RIA analysis of the tissue oxytocin content showed that luteal oxytocin concentrations declined during luteolysis to reach basal values at about the time of the next ovulation. Oxytocin was first measurable in the walls of 3 out of 6 preovulatory follicles during the LH surge, with a small increase in concentration to 26.1 +/- 6.6 pg/mg before ovulation, and a further increase in the young corpus luteum to concentrations exceeding 1 ng/mg 2-3 days later. After the LH surge, oxytocin was also found in the follicular fluid at a concentration of 3.4 +/- 0.3 ng/ml. Using immunocytochemical techniques, oxytocin and neurophysin were first detected in the follicle wall immediately before ovulation, and were localized in the granulosa cells. After ovulation the stained cells initially formed strands which appeared to break down to clusters and then to individual cells as the corpus luteum matured. The immunocytochemical picture also suggested that neurophysin immunoreactivity increased within a few hours of ovulation but that processing to oxytocin may be delayed. Measurements of circulating oxytocin concentrations revealed a pulsatile release pattern throughout the follicular phase with the height of the pulses decreasing from 25 +/- 5 pg/ml during luteolysis to a minimum of 11 +/- 2 pg/ml during the LH surge.
Biosynthesis of oxytocin in the corpus luteum.
In this report we demonstrate that ovine and bovine luteal cells synthesise oxytocin by way of a precursor protein similar to that found in the hypothalamus. Isolated ovine or bovine luteal cells were incubated for up to 12 h with [35S]cysteine. Neurophysin-Sepharose column separation and HPLC of cell extracts demonstrated the presence of [35S]oxytocin. Incorporation of [35S]cysteine was confirmed by performic acid oxidation. Immunoprecipitation of cell extract with anti-rat oxytocin-neurophysin followed by SDS-PAGE yielded 2 radioactive bands of 14 kDa and 11-12 kDa. Immunoprecipitation with anti-oxytocin yielded 1 band at 14 kDa. On SDS-PAGE the 14 kDa band had a similar mobility to rat-hypothalamic oxytocin precursor.
Identification of oxytocin and vasopressin in the testis and in adrenal tissue.
Oxytocin, vasopressin and neurophysin-like immunoreactivity have been identified and measured by radioimmunoassay in extracts of human and rat testis and human fetal adrenal tissue. The authenticity of these polypeptides has been confirmed by their behaviour on high performance liquid chromatography. The concentrations of the hormone were too great to be explained by known circulating levels of the polypeptides, and their presence in steroid secreting organs suggests a possible role for them in steroidogenesis. The peptides may be taken up and concentrated by the tissues but the co-localisation of neurophysins with the hormones points towards local synthesis.
Gonadal sources of the posterior pituitary hormones.
Explore the source record for details and available documents.
Characterization of oxytocin, vasopressin, and neurophysin from the bovine corpus luteum.
Acid extracts of corpora lutea collected from nonpregnant cows were found to contain oxytocin, arginine vasopressin, and neurophysin. The inhibition curves of the oxytocin and vasopressin extracts showed parallelism with the appropriate standard preparations in specific RIAs and eluted at the same position as the standards using high performance liquid chromatography (HPLC). The neurophysin extract showed parallelism in a bovine neurophysin I RIA and had a similar elution position to the standard on both Sephadex G-50 and HPLC. However, its immunoreactive profile on HPLC differed slightly from that obtained with hypophyseal bovine neurophysin I. In nonpregnant cows the oxytocin content (about 1 microgram g-1 wet wt of tissue) was three orders of magnitude greater than the vasopressin content. Levels of luteal oxytocin were considerably lower in pregnant animals. These results show that the bovine ovary is a rich source of neurohypophysial peptides and suggest that oxytocin biosynthesis may occur within the corpus luteum.
Partial reversal of the myometrial progesterone 'block' in the non-pregnant ewe in vivo by oestradiol-17 beta.
In ovariectomized non-pregnant ewes myometrial activity, which was suppressed by continued injection of progesterone, showed a significant recovery of both spontaneous activity and oxytocin responsiveness by 12 and by 24 h after intrauterine administration of oestradiol-17 beta. No such recovery occurred in ewes given vehicle only into the uterus. The oestrogen-induced recovery was lost by 60 h after treatment. Infusion of PGF-2 alpha into the uterine lumen instead of oestrogen provided no support for the possibility that the oestrogen-induced recovery was mediated through PGF-2 alpha secretion. It is suggested that oestradiol may participate in the mechanism of parturition in sheep by accelerating myometrial recovery from the effects of progesterone.
Oestradiol-17 beta both inhibits and stimulates myometrial activity in ewes in vivo.
Spontaneous intrauterine pressure cycles were abolished for periods of several hours in ovariectomized non-pregnant ewes within 8 h of an injection of 50 micrograms oestradiol-17 beta. Following the quiescent period, a phase of intense uterine activity ensued for several hours before again being replaced by several hours of quiescence. This active/inactive cycle disappeared after 2-4 days if further daily injections of oestradiol were not given. During the quiescent periods the uterus retained its responsiveness to oxytocin and PGF-2 alpha. Increasing the dose of oestradiol failed to prolong the periods of quiescence.
Changes in relaxin production by the theca during the preovulatory period of the pig.
Theca and granulosa layers were isolated from the preovulatory follicles of prepubertal gilts which were untreated (Group A), killed 72 h after 1000 i.u. PMSG (Group B), killed 84 h after PMSG (Group C), or killed 84 h after PMSG + 500 i.u. hCG given at 72 h (Group D). The tissues from individual follicles were cultured for 24 h alone (C), with FSH (F) or with LH (L), and the content of immunoreactive relaxin in the culture media was measured by RIA. Concentrations of relaxin-like material were close to the limit of detection of the assay in all granulosa cell cultures, and in the thecal cultures from the untreated gilts. However, thecal cultures from all 19 treated gilts produced relaxin. The mean +/- s.e.m. concentrations (pg/follicle) in Groups AC, BC, CC and DC were 26 X 5 +/- 3 X 04, 93 X 1 +/- 4 X 6, 138 +/- 16 X 4 and 285 X 6 +/- 54 X 1 respectively. Therefore relaxin production was stimulated by PMSG (P less than 0 X 05), with hCG treatment in vivo leading to a further significant increase (P less than 0 X 05). In-vitro treatment with gonadotrophins had no effect in Groups A, C and D, but in Group-B gilts LH produced a significant (P less than 0 X 05) rise in relaxin levels. These studies indicate that the theca is the principal source of relaxin in the porcine preovulatory follicle. The increased production before ovulation suggests that relaxin may be involved in follicular growth or rupture.
On the paper of J. Herczeg [Acta Physiol. Acad. Sci. Hung. 58, 147-155 (1981)].
Explore the source record for details and available documents.
Neurohypophysial hormones in the human ovary.
Explore the source record for details and available documents.
Unsolved problems of relaxin's physiological role.
Explore the source record for details and available documents.
The timing and possible mechanisms of cervical opening in the oestrous rat.
The time that the retention of luminal fluid begins and its release from the uterus in pro-oestrous rats was determined precisely using a dye injection technique. Oestradiol was found to cause dye retention in ovariectomized rats when provided continuously from an implant of 5 mg. Injections of 0.5 microgram/d X 4 were ineffective. Administration or progesterone to rats bearing implants of oestrogen interfered with the ability of the uterus to retain dye. Treatment with relaxin for 28 h prevented retention of eye in pro-oestrous rats whereas treatment at the time of dye injection was without effect. Mechanical stimulation of the cervix did not cause loss of injected dye. It is suggested that the mechanisms of opening of the cervix at coitus and at the end of oestrous may differ.