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E. Decuypere and E. R. Kühn

ABSTRACT

The influence of an intravenous injection of ovine prolactin on the liver monodeiodinase activity and serum concentrations of thyroid hormones and corticosterone was studied in Hisex embryos and chicks after hatching. An injection of 1 and 10 μg ovine prolactin into 18-day-old chick embryos increased serum concentrations of tri-iodothyronine (T3) five-and eightfold respectively after 2 h. At the same time serum concentrations of thyroxine (T4) and reverse T3 (rT3) were decreased in the chick embryo, but only with 10 μg prolactin. This was accompanied by a doubling of the liver monodeiodinase activity. Serum concentrations of corticosterone, however, were not influenced by the prolactin injections. In the 5-day-old chicken, serum concentrations of T3, rT3, T4 and liver T4-5′-monodeiodinase activity were not influenced by 1 or 10 μg prolactin. Serum concentrations of corticosterone after injection of 1 or 10 μg prolactin were doubled compared with controls. These results are compatible with a prolactin-induced shift from a T4-5-monodeiodination into a T4-5′-monodeiodination in the liver at the end of incubation. This effect, however, is not mediated through a prolactin-induced corticosterone release.

J. Endocr. (1985) 104, 363–366

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K Reiprich, E Mühlbauer, E Decuypere and R Grossmann

Abstract

In this study both sexes of two strains of chicken with genetically different growth potentials (broiler- and laying-type) were used to investigate growth hormone (GH) gene expression during posthatch development from day 7 (D7) to D56 by using the in situ hybridization technique and Northern analysis. In pituitaries of both strains a high GH mRNA signal was found as early as D7 by in situ hybridization, showing clear differences in the pattern of gene expression between the two strains. By Northern hybridization sex differences were detectable in all age groups of broilers, with higher levels throughout in males. In layers, however, females showed consistently higher levels compared with males until D21. While signal intensities decreased in the broiler strain during the investigation period, the layer-type strain seemed to express GH mRNA more continuously, reaching significantly (P<0·01) higher GH mRNA levels than broilers at D56. Plasma GH concentrations ran parallel to GH mRNA in early stages but showed a peak earlier at D14 and decreased after D35 in both sexes and strains. Determination of growth as weekly weight gains, however, proved that a period of rapid growth (at a higher level in both sexes of the broiler strain) at D7 was followed by a strong decrease from D14 to D21. A plateau of constant growth was reached until the end of the observation period with similar rates in both strains and sexes. Analysis of plasma thyroid hormones tri-iodothyronine/thyroxine (T3/T4) showed an increase in T3 concentrations in both strains and sexes in early stages and a decrease thereafter. No clear strain differences were measured. T4 plasma concentrations increased from D7 to D14 in broilers and D21 in layers when a plateau was reached. From the results we conclude that generally there is a good correlation between GH mRNA and plasma GH concentrations in both strains investigated. Neither parameter, however, is coupled directly with the growth rate. Thus the early rapid growth corresponds to relatively low levels of GH mRNA and plasma GH concentrations, but high T3 levels. Later, decreased growth rates are linked to increasing amounts of GH mRNA as well as increasing plasma GH concentrations in both layers and broilers. Towards the end of the observation period there was a strain divergence visible with increased amounts of GH mRNA in layers but a strong reduction in broilers. Moreover, plasma GH concentrations decreased more slowly in layers than in broilers.

Journal of Endocrinology (1995) 145, 343–353

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E. J. Nouwen, E. Decuypere, E. R. Kühn, H. Michels, T. R. Hall and A. Chadwick

ABSTRACT

Serum concentrations of arginine vasotocin (AVT), mesotocin and prolactin were determined by radioimmunoassay in Rhode Island Red chickens during and after dehydration, haemorrhage and oviposition. During dehydration increased circulating levels of AVT, mesotocin and prolactin were found. As water deprivation proceeded, marked differences were observed. After an initial rise in serum AVT, mesotocin and prolactin levels during mild and moderate dehydration, concentrations of both AVT and prolactin tended to normalize during continued water deprivation, while those of mesotocin remained high throughout the whole dehydration experiment with the highest at the end of the water-deprivation period. Removal of 5 ml blood at intervals of 10 min during six consecutive time-periods did not affect serum osmolality and circulating levels of AVT and prolactin, but slightly increased mesotocin. These results suggest an osmoregulatory role for AVT and prolactin, whereas mesotocin may be involved in volume control. Finally, 1 min after oviposition, control values of 19·5 ±3·4 pmol AVT/1 (n=9) were raised more than sevenfold to 142·9±12·5 pmol/l (n=11). Thereafter, a decline occurred with a half-life for AVT of 13 min with raised serum levels up to 31 min after oviposition. In contrast, the serum concentrations of mesotocin and prolactin remained unaffected by oviposition.

J. Endocr. (1984) 102, 345–351

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E. R. Kühn, P. Van Osselaer, O. Siau, E. Decuypere and A. Moreels

ABSTRACT

Lambs originating from Suffolk, Milksheep and Texel crossbreeds were injected with saline, 500 μg ovine prolactin or 500 μg ovine GH within 30 min of parturition (n = 10). Birth weight was negatively correlated with plasma concentrations of reverse tri-iodothyronine (rT3) but not with thyroxine (T4), free T4 (expressed as the free thyroxine index) or cortisol. At birth, T3 and T4 plasma concentrations were high and remained high during the 3-h observation period. After 3 h a significantly lower rT3 concentration was found. The cortisol concentration at birth was also high (100–400 nmol/l) but decreased rapidly to basal values after 1 h. An injection of 500 μg prolactin after parturition did not influence the hormonal parameters studied except for rT3 where, after 2 h, lower plasma concentrations were found compared with controls.

Growth hormone raised T3 levels from 4·80 ± 0·44 (s.e.m.) nmol/l at birth to 6·74 ± 0·42 nmol/l at 1 h after birth (P < 0·01) and to 6·51 ± 0·42 nmol/l after 2 h (P < 0·05). At both times these values were significantly (P < 0·001) different from saline-injected controls. GH decreased rT3 from 6·77 ± 0·71 nmol/l at birth to 5·42 ± 0·54 nmol/l after 1 h (P < 0·05) and to 5·10 ± 0·45 nmol/l after 2 h (P < 0·01; values were also significantly different from saline controls at P < 0·05 and < 0·005 respectively). At the same time total and free T4 concentrations were increased. No influence of prolactin or GH injection on plasma cortisol concentrations was seen. It is concluded that GH may play an important role in the maturation of thyroid functions during the perinatal period of lambs by increasing T4 secretion and by increasing the conversion of T4 to T3 and decreasing the T4 to rT3 conversion rates.

J. Endocr. (1986) 109, 215–219

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R. Peeters, N. Buys, D. Vanmontfort, J. Van Isterdael, E. Decuypere and E. R. Kühn

ABSTRACT

The influence of TRH and TSH injections on plasma concentrations of tri-iodothyronine (T3) and thyroxine (T4) was investigated in neonatal (injection within 0·5 h after delivery) and growing lambs and in normal, pregnant and lactating adult ewes (all 2 years old and originating from Suffolk, Milksheep and Texal cross-breeds). Neonatal lambs had higher levels of T3, T4 and GH compared with all other groups, whereas prolactin and TSH were higher in lactating ewes. In all animals, injections of TRH increased plasma concentrations of prolactin and TSH after 15 min but not of GH at any time. Small increases in T3 and T4 were observed in neonatal lambs, without any effect on the T3 and T4 ratio, after prolactin administration, whereas prolactin did not influence plasma concentrations of T3 or T4 in all other experimental groups. Similar results for thyroid hormones were obtained after TRH or TSH injections. It was therefore concluded that the effects observed after TRH challenge were mediated by the release of TSH. With the possible exception of neonatal lambs, plasma concentrations of T3 after administration of TRH or TSH were always increased before those of T4; the increase in T3 occurred within 0·5–1 h compared with 2–4 h for T4 in all experimental groups. This resulted in an increased ratio of plasma T3 to T4 up to 4 h after injection. It is concluded that, in sheep, TRH and TSH preferentially release T3 from the thyroid gland probably by a stimulatory effect of TSH on the intrathyroidal conversion of T3 to T4.

Journal of Endocrinology (1992) 132, 93–100

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A. Vanderpooten, V. M. Darras, L. M. Huybrechts, P. Rudas, E. Decuypere and E. R. Kühn

ABSTRACT

The effects of hypophysectomy on GH binding to liver membranes of young chicks were studied 3 days and 1 week after surgery. Specific binding of 125I-labelled chicken GH (cGH) to MgCl2-treated liver microsomal fractions of hypophysectomized animals was two- to fivefold greater than to those of sham-operated or control (non-operated) birds. This effect was due to a rise in binding capacity rather than a change in binding affinity of the GH receptor. Two daily injections of cGH (20 μg/animal) returned the number of hepatic GH receptors from hypophysectomized chicks to the level of the sham-operated ones. Administration of GH to the latter group did not cause a significant lowering of specific binding or number of receptors. No positive correlation between GH binding and plasma concentrations of insulin-like growth factor-I (IGF-I) was observed; although GH binding increased, IGF-I levels were lower for the hypophysectomized group. Since the number of hepatic GH receptors and the plasma GH levels were inversely correlated, it was concluded that the GH receptors in the liver of the chicken can be down-regulated by GH. This possibly explains why GH binding is low in posthatch and young chicks, because circulating GH concentrations are high during this period.

Journal of Endocrinology (1991) 129, 275–281

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J. Buyse, E. Decuypere, P. J. Sharp, L. M. Huybrechts, E. R. Kühn and C. Whitehead

ABSTRACT

Daily changes in the concentrations of plasma corticosterone, prolactin, thyroid hormones and somatomedin C were measured in 28-day-old fat and lean lines of broilers produced by selection for high and low concentrations of very low density lipoproteins (VLDL). The effects of daily injections of corticosterone on the concentrations of these hormones and on fattening were compared in the two lines. The selection procedure had no effect on the concentrations of any of the hormones. However, daily rhythms in concentrations of plasma corticosterone, tri-iodothyronine (T3) and prolactin were less often observed in the fat line than in the lean line. No differences were seen between lines in the daily rhythms in plasma thyroxine (T4) and somatomedin C. Daily injections of 2500 μg corticosterone/kg body weight, in both lines, depressed mean concentrations of plasma prolactin, T3 and somatomedin C and body weight. This dose of corticosterone also increased abdominal fat pad and liver weights expressed as a percentage of body weight. The liver and fat pad responses to 2500 μg corticosterone in both lines were greater when the steroid was injected at the end rather than towards the beginning of the 14-h daily photoperiod. There was no difference between the lines in the fattening response to corticosterone. Lower doses of 100 and 500 μg corticosterone per day did not induce fattening or affect concentrations of plasma prolactin. They did, however, depress concentrations of plasma T3. Concentrations of plasma T4 were increased in both lines treated with 2500, but not with 100 or 500 μg corticosterone, towards the beginning of the daily photoperiod. It is concluded that selection for low and high concentrations of VLDL does not affect mean levels of the hormones measured at 28 days of age or the fattening response to corticosterone. It does, however, alter the amplitude in the daily rhythm in concentrations of corticosterone, T3 and prolactin. The fattening response to corticosterone is associated with depressed concentrations of plasma prolactin, T3 and somatomedin C.

J. Endocr. (1987) 112, 229–237

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E. R. Kühn, E. Decuypere, K. Hemschoote, L. Berghman and J. Paulussen

The influence of an intravenous injection of ovine prolactin on circulating levels of thyroid hormones was studied in Rhode Island Red embryos and chicks after hatching. In the chick embryo, 2 h after injection of 0·1 μg prolactin (on incubation day 19), serum tri-iodothyronine (T3) increased threefold; after 10 or 100 μg prolactin (on incubation day 18) serum T3 increased 15- to 25-fold. These profound increases were not observed in chicks after hatching. Serum concentrations and thyroid content of thyroxine (T4) in embryos and chicks of all ages studied were not influenced by the prolactin injections. Maximal serum concentrations of reverse T3 (rT3) were found on incubation day 18 (110·25 ± 23·36 pmol/l; 71·66 ± 15·18 pg/ml; n = 8), whereas after hatching no rT3 could be detected. An injection of 10 μg prolactin on day 18 depressed serum rT3 after 2 h to 5·68 ± 3·20 pmol/l (3·69 ± 2·08 pg/ml; n = 8; P<0·001); the effect of 100 μg prolactin was less pronounced. After hatching, chronic administration of prolactin resulted in decreased serum levels of T3, but not of T4, and hypertrophy of the follicles in the thyroid gland. It is concluded that prolactin plays a major role in the maturation of embryonic thyroid metabolism by changing the T4-5-monodeiodination into a T4-5′-monodeiodination. The hypertrophy of the thyroid gland observed after hatching following prolonged prolactin administration may be due to decreased negative feedback of T3 on the hypophysis.

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L. R. Berghman, J. van Beeumen, E. Decuypere, E. R. Kühn and F. Vandesande

ABSTRACT

The immunization of mice with an affinity-purified glycoprotein preparation from chicken pituitary tissue yielded several monoclonal antibodies towards the recently described glycosylated variant of chicken GH. As all these antibodies recognize the classical (non-glycosylated) GH molecule equally well, they provide a suitable tool for the development of both a specific immunoadsorbent and an assay method. This paper deals with the surprising purification power of the immunoadsorbent that was produced with one of the monoclonal antibodies. The resulting preparation was more than 99% pure as assessed by reversed phase high-performance liquid chromatography and sodium dodecyl sulphate-polyacrylamide gel electrophoresis, so that no further purification steps were needed before the determination of the amino acid sequence of the material. The efficiency of the purification protocol as determined by a homologous, monoclonal antibody-based radioimmunoassay was virtually absolute. Moreover, the affinity-purified GH preparation was a mixture representing the multiple molecular forms of pituitary chicken GH, including both oligomeres and glycosylated GH. The purified preparations were finally used to demonstrate the hepatic 5′-monodeiodinase-stimulating activity of GH in the chicken embryo (results not shown), in order to prove that the biological activity of the molecule had not been damaged by elution from the immunoadsorbent.

J. Endocr. (1988) 118, 381–387

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M M Cokelaere, P Busselen, G Flo, P Daenens, E Decuypere, E Kühn and M Van Boven

Abstract

Simmondsin, a glycoside extracted from jojoba meal (Simmondsia chinensis), causes a reduction in food intake after oral administration. To investigate the mechanism by which simmondsin reduces food intake, fasted and freefeeding rats were given simmondsin-supplemented food and simultaneously injected with devazepide, a specific antagonist of peripheral-type cholecystokinin receptors (CCKA receptors). In free-feeding rats, supplementation of food with 0·5% simmondsin caused a reduction in food intake of ± 40% in the period of 4 h following food presentation. Intraperitoneal injection of 100 μg devazepide/kg body weight prevented this effect. In rats fasted for 20 h, the food intake in the 30 min after presentation of food supplemented with 0·15% or 0·50% simmondsin was reduced in a dose-related manner; this was also inhibited by simultaneous application of devazepide. It is suggested that peripheral CCKA receptors are involved in the effect of simmondsin on food intake. However, a direct effect of simmondsin on CCKA receptors has been excluded, since simmondsin was unable to cause contraction of the guinea-pig gallbladder in vitro.

Journal of Endocrinology (1995) 147, 473–477