The Journal of endocrinology.
Publisher:
Society for Endocrinology. Bristol, UK : BioScientifica (2011)
Frequency: Monthly
Country: England
Language: English
Author(s):
Society for Endocrinology.
Start Year:1939 -
ISSN:
0022-0795 (Print)
1479-6805 (Electronic)
0022-0795 (Linking)
1479-6805 (Electronic)
0022-0795 (Linking)
Impact Factor
4
2022
| NLM ID: | 0375363 |
| (DNLM): | J19940000(s) |
| (OCoLC): | 01754564 |
| Coden: | JOENAK |
| LCCN: | 64005925 |
| Classification: | W1 JO642 |
The effects of corticotrophin-releasing hormone, arginine vasopressin and their antagonists on ACTH release from perifused horse anterior pituitary cells. Antagonists are useful for probing hormone action and receptor characteristics. In this study we have investigated the inhibitory effects of analogues of arginine vasopressin (AVP) and corticotrophin-releasing hormone (CRH) on stimulated release of immunoreactive ACTH from perifused equine anterior pituitary cells in vitro. Our aims were to gain some insight into the characteristics of the CRH and AVP receptors of the horse pituitary and to establish whether the response induced by AVP and CRH together could be blocked by combining antagonists. Experimental design included 5-min pulses of AVP ...
Prepartum adrenocortical maturation in the fetal foal: responses to ACTH. The present study was carried out on 19 chronically catheterized mares and fetuses in late gestation (term > 320 days). In six animals which were monitored up to the time of delivery of a live foal, plasma and amniotic fluid cortisol concentrations remained low until 4-5 days before parturition when there was a rapid, significant rise (P < 0.05) which was not accompanied by any corresponding changes in maternal plasma cortisol. Circulating fetal ACTH concentrations became more variable close to delivery and ANOVA revealed no significant increases during this critical period, although a n...
Effects of the opioid antagonist naloxone on release of luteinizing hormone in mares during the anovulatory season. To investigate an involvement of endogenous opioids in the regulation of circannual changes in reproductive activity, effects of the opioid antagonist naloxone on the concentration of immunoreactive and bioactive luteinizing hormone (LH) in plasma were measured in mares during the anovulatory season. Naloxone (0.5 mg/kg i.v.) caused a significant increase (P < 0.05) in immunoreactive as well as bioactive LH concentration in plasma. The amplitude of the increase in LH concentrations measured with an in vitro bioassay was more pronounced than the amplitude of the increase in LH secretion dete...
The dynamics of gonadotrophin-releasing hormone, LH and FSH secretion during the spontaneous ovulatory surge of the mare as revealed by intensive sampling of pituitary venous blood. Conflicting views exist on the mode of gonadotrophin-releasing hormone (GnRH) secretion during the ovulatory LH surge and the relative importance of changes in pituitary responsiveness to GnRH in generating the LH surge. This disagreement may stem from species differences and/or methodological problems. To provide data on the exact relationship between GnRH and gonadotrophin secretion during the spontaneous LH surge, we collected pituitary venous (PV) blood every 30 s for 3-4 h from eight mares and then assayed GnRH (in six of the mares), FSH and LH. Jugular blood was also collected from twelv...
Growth hormone secretion in the horse: unusual pattern at birth and pulsatile secretion through to maturity. A heterologous radioimmunoassay was developed and validated for the measurement of horse GH in plasma. It utilized recombinant-derived bovine GH as the radiolabelled ligand, a guinea-pig anti-porcine GH serum as first antibody and pituitary-derived horse GH as standard. Cross-reactivities were high with all of the pituitary and recombinant-derived GH preparations tested (49-140%) and very low (< 0.3%) with horse FSH, LH and prolactin. A synthetic analogue of GH-releasing factor(1-29) stimulated the expected pattern of GH release in foals. Plasma GH concentrations in foals were low at birth (< ...
The effect of cortisol on the secretion of ACTH by anterior pituitary cells of the horse in culture. Perifused equine anterior pituitary cells were used to investigate the effect of cortisol on the ACTH response to pulses of corticotrophin-releasing hormone (CRH; 0.01 nmol/l) and arginine vasopressin (AVP; 100 nmol/l), given for 5 min every 30 min for 690 min and ACTH measured in 5-min fractions. At the fourth pulse of secretagogue (0 min), a constant perifusion with cortisol began (0 nmol/l (control), 100, 200, 500, 5000 and 50,000 nmol/l) and continued until the ninth pulse (150 min). For each pulse of secretagogue, the amount of ACTH (pmol) secreted in response to each pulse (ACTH response...
Factors affecting ACTH release from perifused equine anterior pituitary cells. The multifactorial control of ACTH is well established. We wished to establish and characterize an in-vitro perifusion system, using equine anterior pituitary cells and physiological concentrations of secretagogues, to investigate factors which affect the dynamics of ACTH secretion. Anterior pituitary tissue was divided for dispersion into cells with collagenase, trypsin or dispase, or by mechanical dispersion. After dispersal followed by 18-h incubation, cells were perifused and the ACTH response to 10-min pulses of arginine vasopressin (AVP; 100 nmol/l), corticotrophin-releasing hormone (CRH...
Concentrations of arachidonate metabolites, steroids and histamine in preovulatory horse follicles after administration of human chorionic gonadotrophin and the effect of intrafollicular injection of indomethacin. This study investigated the sequence of hormonal changes within the preovulatory follicles of mares. Mares were injected i.v. with 2500 IU human chorionic gonadotrophin (hCG) when a preovulatory follicle of 35 mm in diameter was detected. Fluid was aspirated from preovulatory follicles before (0 h), and 12, 24 and 36 h after administration of hCG. Concentrations of progesterone, prostaglandin (PG) E2, PGF, 6-keto-PGF1 alpha and thromboxane B2 in follicular fluid increased significantly (P less than 0.01) between 0 and 36 h. At 36 h, PGE2 was present in highest concentrations, followed by PGF a...
Radioimmunoassay of inhibin in various mammals. A sensitive radioimmunoassay (RIA) for the determination of inhibin in peripheral plasma and tissue homogenates of different species has been developed using antisera to partially purified bovine follicular fluid (bFF) inhibin and 125I-labelled bFF 32 kDa inhibin. Antisera were produced by immunization of rabbits with partially purified bFF inhibin prepared by immunoaffinity chromatography. Increasing doses of a high titre antiserum could neutralize the suppressing effect of bFF, porcine follicular fluid and rat ovarian homogenate on FSH secretion from rat anterior pituitary cells in culture. ...
Steroids in equine testes: the identification of endogenous 19-hydroxy and 19-nor neutral steroids by gas chromatography–mass spectrometry. After homogenization of testicular tissue from stallions aged 1, 2 and 5 years, the unconjugated and conjugated steroids were isolated by a combined solvent-solid extraction procedure. The conjugates were further separated into glucuronides and sulphates by chromatography using Sephadex LH-20. After enzyme hydrolysis and solvolysis of the respective conjugate classes, the three extracts, unconjugated steroids, aglycones and solvolysed sulphates, were purified by chromatography using Kieselgel 60H columns. Five fractions were resolved from each extract; an aliquot of each fraction was derivatiz...
Characteristics of milk ejection, associated intramammary pressure changes and oxytocin release in the mare. The neuroendocrine reflex theory of milk ejection was investigated in the horse under natural suckling conditions. To this end 12 lactating mares were provided with acute jugular catheters and with intramammary pressure (IMP) recording catheters. The foal had free access to the contralateral mammary complex. Intramammary pressure could thus be recorded while blood was drawn simultaneously for oxytocin analysis from the undisturbed animal. Suckling periods associated with a characteristic increase in IMP lasted significantly longer than unsuccessful nursing attempts. Elements of successful suck...
Secretion rates and short-term patterns of gonadotrophin-releasing hormone, FSH and LH in the normal stallion in the breeding season. Pituitary venous blood was collected by a painless nonsurgical cannulation method from five ambulatory stallions at 5-min intervals for 5-6 h during the breeding season. In four adult stallions, statistical analysis showed that pulses of gonadotrophin-releasing hormone (GnRH) and LH were coincident (P less than 0.01), as were pulses of FSH and LH (P less than 0.05). Furthermore, the patterns of changes in concentration of FSH and LH were highly correlated in each of the four stallions. However, seemingly ineffective pulses of GnRH were also observed, with 28% of GnRH pulses failing to induce a...
Effect of isolation stress on concentrations of arginine vasopressin, alpha-melanocyte-stimulating hormone and ACTH in the pituitary venous effluent of the normal horse. A non-surgical, non-stressful technique was used for collection of pituitary venous blood from five conscious horses every minute for two 10-min periods before and during isolation from the herd, which caused a predictable, yet humane and physiological, emotional stress. Pituitary blood was also sampled every 5 min for two approximately 90-min periods before and after isolation, while jugular blood was sampled every 15 min throughout the experiment. During isolation, all horses became agitated, hyperventilating and sweating. Packed red cell volume increased, as did pituitary venous concentrati...
Nucleotide (cDNA) sequence encoding the horse gonadotrophin alpha-subunit. Several cDNA clones corresponding to mRNA for the alpha-subunit of the horse (Equus caballus) pituitary and placental (chorionic) gonadotrophic hormones have been isolated and sequenced. Polyadenylated mRNA was purified from horse pituitary glands (the source of FSH and LH) and horse placental tissues (the source of chorionic gonadotrophin; CG). The mRNA preparations were characterized by in-vitro translation and Northern hybridization techniques using human and ovine gonadotrophin cDNA clones as probes. Complementary DNA libraries were created from the pituitary and placental mRNAs and a huma...
Secretion rates and short-term patterns of gonadotrophin-releasing hormone, FSH and LH throughout the periovulatory period in the mare. We have developed a non-surgical technique for long-term collection of pituitary venous blood which consists of slightly diluted hypophysial portal blood into which pituitary hormones have been secreted. In these experiments jugular and pituitary venous blood samples were collected from five unmedicated, ambulatory mares at 5-min intervals for 2-6 h on 11 occasions during the 6 days surrounding the ovulatory LH peak. Jugular blood only was collected from another five periovulatory mares without pituitary cannulae. The duration of oestrus was similar in mares with and without pituitary cannulae...
Influence of administration of ovarian steroids on the function of neutrophils isolated from the blood and uterus of ovariectomized mares. The function of blood and uterine luminal neutrophils from ovariectomized mares treated with ovarian steroids was investigated 18 h after intrauterine infusion of 1 X 10(9) Streptococcus zooepidemicus. Random migration of blood neutrophils under agarose was reduced by treatment with progesterone compared with that of neutrophils from oestradiol-treated and control mares. In-vitro addition of progesterone to blood neutrophils from acyclic ponies also reduced migration. Uterine neutrophils did not migrate under agarose which was probably an effect of bacterial phagocytosis. Hormone treatment had...
Effect of graded doses of gonadotrophin-releasing hormone on serum LH concentrations in mares in various reproductive states: comparison with endogenously generated LH pulses. Luteinizing hormone release induced by a range of small (3.3-33 micrograms) and large (300-500 micrograms) i.v. doses of gonadotrophin-releasing hormone (GnRH) was measured in acyclic (n = 4), luteal phase (n = 3) and follicular phase (n = 5) mares and compared with endogenously generated LH pulses in the same reproductive states. Extrapolation from log-linear dose-response curves showed that an LH pulse comparable to an endogenous one would be simulated by i.v. injection of 7.0 (n = 4) and 4.1 (n = 6) micrograms GnRH in luteal and follicular phase mares respectively; a much smaller dose than ...
Identification of 3 beta-hydroxy-5,7-androstadien-17-one as a secretory product of the fetal horse gonad in vivo and in vitro. Isolation of 3 beta-hydroxy-5,7-androstadien-17-one, as a major component of steroids extracted from vein blood of the fetal gonads of the horse, supports the proposed role for the compound as a precursor for equilin formation in the placenta of the mare. The 5,7-diene was extracted from blood collected from gonadal veins of fetal ovaries and testes in situ, and from a fetal testis connected to an artery in the neck region of the mare. Perfusion of fetal gonads in the laboratory was carried out to allow longer periods of collection. In addition, isolated cell preparations from a fetal testis w...
Effects of dietary energy supply on serum thyroxine, tri-iodothyronine and insulin concentrations in young horses. The effects of meal ingestion on the circulating concentrations of the growth-regulating hormones thyroxine (T4), tri-iodothyronine (T3) and insulin were examined in weanling Thoroughbreds fed 70% (diet A), 100% (diet B) or 130% (diet C) of their energy and protein requirements. Peak insulin concentrations occurred 1, 2 and 3 h after the ingestion of diets C, B and A respectively. Increases in plasma glucose concentrations preceded the increases in serum insulin concentrations. Serum T4 concentrations increased after the ingestion of diets A and B and decreased after diet C. In contrast, serum...
Changes in plasma cortisol concentrations during the ovulatory cycle of the mare. Daily blood samples from four mares were assayed for cortisol through a total of eight ovulatory cycles. Mean cortisol concentrations on days -14, -13, -10, -9 and -8 before ovulation (dioestrus) were greater than on days -5 to -1 (oestrus). The highest mean (+/- S.E.M) value of cortisol occurred on day -10 (260 +/-28 nmol/l) and the lowest on day -2 (142 +/- 14 nmol/l). A single episode on a day in late dioestrus characterized the maximum cortisol value per cycle for five of eight cycles. Extraction of plasma samples with petroleum ether or chromatography before assay, to eliminate interferen...
The use of heterologous radioimmunoassays for the measurement of follicle-stimulating hormone and luteinizing hormone concentrations in horse and donkey serum. Heterologous double-antibody radioimmunoassay were developed for the measurement of FSH and LH concentrations in the serum of both horses and donkeys. The FSH assay employed a rabbit anti-ovine FSH serum which showed a complete lack of cross-reaction with equine chorionic gonadotrophin (eCG) and negligible cross-reaction with equine LH. The LH assay utilized an antiserum raised against highly purified eCG. This similarly showed negligible cross-reaction with equine FSH but its high cross-reactivity with eCG prevented the measurement of equine LH concentrations in serum when eCG was also presen...
Identification of 3 beta-hydroxy-5,7-pregnadien-20-one and 3 beta-hydroxy-5,7-androstadien-17-one as endogenous steroids in the fetal horse gonad. The 5,7-dienes, 3 beta-hydroxy-5,7-pregnadien-20-one and 3 beta-hydroxy-5,7-androstadien-17-one were extracted from fetal horse gonads and purified by solvent partition, thin-layer chromatography and high performance liquid chromatography. The isolated steroids were identified by comparison with the synthetic steroids using ultraviolet and mass spectroscopy and by gas chromatography-mass spectroscopy. The identification of these compounds as endogenous steroids, together with the data on their biosynthesis reported previously, support the proposal that in the fetal horse gonad there is a 5,7-d...
Hormonal changes in the immature rat after administration of pregnant mare serum gonadotrophin: influence of body weight. We have shown previously that pregnant mare serum gonadotrophin (PMSG) induces ovulation only in rats weighing over 60 g on the day of injection. The under-60 g rats do not ovulate although they secrete a preovulatory surge of a pleiomorphic form of LH. Presumably this pleiomorph is inactive. Comparisons were made of plasma hormone concentrations in rats treated with PMSG that weighed over and under 60 g. The measurements were made on samples taken between 13.00 and 22.00 h on the day of the expected preovulatory LH surge. Prolactin and corticosterone levels were lower in the lighter group com...
Seasonal and circadian changes of testosterone levels in the peripheral blood plasma of stallions and their relation to semen quality. Three stallions were bled each hour for 25 h at 28-day intervals throughout 1 year. Testosterone levels were pulsatile. Pulse frequency and pulse amplitude were higher in the summer months than at other times (P less than 0.01). The number of testosterone pulses also varied throughout the day, with the greatest frequency occurring in the afternoon (14.00-17.00 h) and at night (22.00-01.00 h). Mean testosterone levels were highest in the summer (P less than 0.01) but showed a secondary, smaller increase in the autumn. Semen characteristics were assessed from measurements of 222 stallions. Semen...
Identification and measurement of testosterone in plasma and follicular fluid of the mare, using gas chromatography-mass spectrometry associated with isotope dilution. Testosterone has been identified by mass spectrometry in blood and follicular fluid aspirated from mature Graafian follicles of mares. Quantitative measurements made by gas chromatography-mass spectrometry have validated the determination of plasma testosterone made by radioimmunoassay. However, because of high levels of epitestosterone (17 alpha-hydroxyandrost-4-en-3-one) in the follicular fluid, radioimmunoassay overestimates the true concentrations of testosterone. The occurrence of testosterone in mare follicular fluid at a concentration which is two orders of magnitude higher than that in...
Pancreatic beta-cell function in the fetal foal and mare. Insulin secretion and the factors influencing beta-cell function were investigated in the chronically catheterized fetal foal and mare during the second half of gestation. The response of the fetal beta cells to exogenous glucose was also examined. The mean concentration of insulin in the fetal foal was 7.5 +/- 0.5 (S.E.M.) microunit./ml (n = 20) which was significantly less than the corresponding maternal value of 49.0 +/- 5.0 microunit./ml (n = 20, P < 0.01). The insulin concentration in non-pregnant horses was 24.5 +/- 1.5 microunit./ml (n = 5) which was significantly less than the value...
Purification and characterization of donkey chorionic gonadotrophin. Serum of the pregnant donkey, like that of the mare, contains a gonadotrophin of chorionic origin. The chorionic gonaditrophin of the donkey (dCG) has been isolated in purified form from the serum of pregnant donkeys using methodology previously employed for the purification of pregnant mare chorionic gonadotrophin (eCG). Unlike eCG, dCG is predominatly an LH in biological tests. In the in-vitro rat Leydig cell assay, dCG was as active as eCG, but in the in-vitro rat seminiferous tubule assay for FSH and in the augmentation assay, dCG was considerably less potent than eCG (1-10%). Specific rat...
Renin, aldosterone and cortisol in the thoroughbred horse. To assess the renin-aldosterone system in a large mammal, simultaneous morning activities of plasma renin and concentrations of aldosterone and cortisol were measured in 25 thoroughbred horses. Renin was relatively low in all horses (0.16 +/- 0.02 (S.E.M.) ng angiotensin I/ml per h), levels of aldosterone in plasma were 527 +/- 130 pmol/1 and levels of cortisol in plasma were 141 +/- 11 nmol/l. Levels of aldosterone were significantly correlated with levels of renin in all horses (r = 0.62, P less than 0.001) but not with those of cortisol, and renin was negatively correlated with age in male ...
Foetal and maternal plasma concentrations of 13, 14-dihydro-15-oxo-prostaglandin F in the mare during late pregnancy and at parturition. The concentrations of 13, 14-dihydro-15-oxo-prostaglandin F(PGFM), the stable metabolite of prostaglandin F, were measured in the plasma of catheterized mares and foetuses and non-catheterized thoroughbred mares and ponies during the last months of gestation. The plasma concentration of PGFM increased gradually towards term in all groups of animals. During the operation for insertion of catheters, maternal and foetal concentrations of PGFM were high, but the values fell to basal levels 24--48 h after the operation. It was found the preoperative starvation (24 h) led to a rise in the concentrat...