Differences in hepatic growth hormone receptor binding during development of normal and dwarf chickens

The aim of this study was to compare the growth hormone (GH) receptor in liver microsomal fractions of normal chickens (Dw) and chickens carrying the dwarf gene (dw). Specific binding of GH to its hepatic receptor was significantly higher for Dw embryos from d 14 till d 20 of incubation than for dw embryos. The difference in binding was due to a decreased binding capacity but not affinity in the livers of the dwarf embryos. The same binding pattern was found in livers of adult chickens: lower binding was again caused by a lower number of GH receptors and at this stage the difference was even clearer than during embryonic development. Binding studies on livers of growing chicks demonstrated that binding was low for both genotypes, but a small though significant difference between them remained. The cause of this decrease in number of GH receptors in dwarf birds has yet to be determined but may be due to the primary action of the dwarf gene.     

A decreased capacity of hepatic growth hormone (GH) receptors and failure of thyrotrophin-releasing hormone to stimulate the peripheral conversion of thyroxine into triiodothyronine in sex-linked dwarf broiler hens

The effect of two different doses of thyrotrophic releasing hormone (TRH) upon the plasma levels of growth (GH) and thyroid hormones in both sex-linked dwarf (dw) and normal (Dw) broiler hens was determined. In normal hens, 1.5 and 24 microg TRH/kg increased the GH plasma concentrations after 15 min. Plasma concentrations of T3 increased significantly 1 h after TRH injection, whereas T4 concentration decreased after 2 following injection of 24 microg/kg TRH. In dwarf hens both doses of TRH increased the plasma concentrations of GH and the GH response lasted longer. However, TRH was ineffective in raising T3 and T4 levels. Saline-injected dwarf birds showed no differences in plasma T4 and T3 levels in comparison with normal hens. A smaller number of hepatic cGH receptors was found in dwarf hens, whereas the affinity of the hepatic GH receptor was not influenced by the genotype. It is concluded that the sex-linked dwarf broiler hen is unable to respond to a TRH-induced GH stimulus probably because of a deficiency in hepatic GH receptors resulting in a failure to stimulate the T4 to T3 converting activity.     

Selenium content of livers from sex-linked dwarf and normal broiler breeders

Plasma concentrations of cGH, T₃, and T₄ were not different between dwarf and normal broiler breeders. Normal hens had a liver selenium content of 710±35 ng/g, and dwarf hens 656 ±nine ng/g (n=8). Following injections into a wing vein of different doses (1.5, 3, 6, 12, and 24 μg/kg) of the hypothalamic hormone TRH, GH was increased after 15 min. This effect seemed to last longer in dwarf chickens. Plasma concentrations of T₃ increased significantly 1 h after TRH in normal hens, but TRH was ineffective in raising T₃ levels in dwarf animals. The selenium content of livers obtained following decapitation after 2 h was also increased in normal hens up to 902±42 ng/g using the highest dose of TRH (24 μg/kg). This seemed not to be the case for dwarf animals. A much smaller. number of hepatic cGH receptors was also found in dwarf hens, whereas the affinity of the hepatic GH receptor was not influenced by the genotype. It is concluded that the sex-linked dwarf hens are unable to increase their hepatic T₄ into T₃ conversion following a TRH challenge probably because of a deficiency in hepatic GH receptors. The lower content of selenium in dwarfs and their inability to increase its uptake after TRH seem therefore to support the hypothesis that selenium has a direct role in the activity of the 5′-deiodinase complex.     

Central somatostatinergic regulation of growth hormone secretion in dwarf chickens.

1. Basal circulating growth hormone (GH) concentrations in sex-linked-dwarf (SLD) chickens were unaffected by the intracerebroventricular (icv) injection of 10, 50 or 100 micrograms somatostatin (SRIF). 2. The GH response to systemic thyrotropin-releasing hormone (TRH; 10 micrograms/kg, iv) was, however, 'paradoxically' enhanced 20 min after icv SRIF administration. 3. A lower dose (1.0 micrograms) of SRIF had no effect on basal or TRH-induced GH release. 4. High-titre SRIF antisera (4 microliters) also had no acute effect on basal plasma GH concentrations, but augmented the GH response to TRH challenge. 5. SRIF would appear to act at central sites to modulate stimulated GH secretion in SLD chickens.     

MicroRNAs expression profiles in adipose tissues and liver from sex-linked dwarf and normal chickens

Recently, increasing evidence suggested that miRNAs contribute to the regulation of adipose deposition and fat metabolism [1-7]. Meanwhile, several studies have consistently indicated that miRNAs are involved in metabolic functions of the liver, including glucose and lipid metabolism [8-12].     

伴性矮小型鸡GH、GHR和IGF-1基因的表达变化

采用荧光实时定量PCR的方法, 从转录水平上分析了伴性矮小型鸡和普通鸡肝脏中GH、GHR和IGF-1基因的表达变化趋势。结果表明：伴性矮小型鸡和普通鸡肝脏组织中GH的mRNA表达量没有明显差异, 而GHR在矮小鸡中的表达量明显比普通鸡的高3倍多, 但IGF-1基因在矮小鸡肝脏中的表达量却远远低于普通鸡, 差异达到2个数量级。这表明, 伴性矮小型鸡GHR外显子10 和3′非翻译区的长片断缺失并没有降低GHR基因的表达, 相反有所增高, 这一过程中可能存在相应的功能代偿机制。与此同时, 在伴性矮小型鸡肝脏中几乎观察不到IGF-1基因的表达, 证明正是由于GHR基因的缺陷影响了GH生理效应的发挥。实验结果印证了伴性矮小表型与GH和GHR的转录水平无关, 而可能是GHR编码产物异常阻碍了GH-GHR-IGF信号通路, 导致IGF-1表达受阻, 不能发挥正常的生理功能。     

Triiodothyronine affects the phytohemagglutinin to concanavalin A proliferative response ratio in sex-linked dwarf chickens

Euthyroid Cornell K strain and sex-linked dwarf (SLD) strain cockerels (which have abnormally low serum triiodothyronine concentrations) were supplemented with either 0, 0.01, 0.1, or 1.0 ppm of triiodothyronine (T3) in the diet. Peripheral blood lymphocytes (PBL) from these cockerels were obtained by slow-speed centrifugation (slow-spin-prepared PBL). The proliferative response of these PBL to phytohemagglutinin (PHA) and concanavalin A (Con A) was determined when the chicks were 6, 9, and 12 weeks of age. Con A responsiveness was also determined in 12-week-old cockerels using PBL which were separated on Ficoll (Ficoll-prepared PBL). Using slow-spin-prepared PBL, PHA, and Con A responsiveness increased in both strains with increasing levels of T3 supplementation. This enhancing effect of T3 was particularly evident in older cockerels. In 6- and 12-week-old SLD strain cockerels, the PHA:Con A response ratio was significantly (P less than 0.05) lower than in K strain cockerels. At 12 weeks of age the PHA:Con A response ratio of the SLD strain was elevated to K strain control levels by T3 supplementation. Therefore, the lower PHA:Con A response ratio in the SLD strain appears to be partially due to the existing peripheral hypothyroidism in this strain. Using Ficoll-prepared PBL, the effects of T3 on Con A responsiveness differed from those observed when slow-spin-prepared PBL were used. From this study we conclude that T3 supplementation affects mitogen responsiveness and the PHA:Con A response ratio. However, the effects of T3 on mitogen responsiveness depend on the age of the chicken, the level of T3 supplemented, the T cell population stimulated, and the method of lymphocyte enrichment.     

Thyroid hormone response to thyrotrophin releasing hormone (TRH) in the sex-linked dwarf chicken

The effect of an injection of thyrotrophin releasing hormone (TRH) on plasma levels of thyroid hormones was studied in dwarf and normal Rhode Island Red chickens with similar genotypes other than for the sex-linked dwarf gene dw. The sex-linked dwarf chickens had different plasma iodothyronine levels from control normal chickens: high thyroxine (T4), low triiodothyronine (T3) and similar reverse T3 (rT3) levels. The injection of TRH (10 micrograms/kg) in 5-day- and 5-week-old normal chickens increased the plasma T4 within 30 min without a significant increase in T3, whereas the injection of TRH in 11-and 26-week-old normal chickens increased plasma T3 60 min later. In dwarfs the response of T4 to TRH was the same as that in normals but no increased T3 response was observed. The plasma level of rT3 was not influenced by the TRH injection in either strain. These results suggest that although in the sex-linked dwarfs thyroidal response to exogenous TRH is similar to that of normals, the dwarf gene dw inhibits the conversion of T4 to T3 in peripheral tissues without any inhibitory effect on rT3 production.     

Enlargement of interscapular brown adipose tissue in growth hormone antagonist transgenic and in growth hormone receptor gene-disrupted dwarf mice

Growth hormone (GH) acts on adipose tissue by accelerating fat expenditure, preventing triglyceride accumulation, and facilitating lipid mobilization. To investigate whether GH is involved in the development and metabolism of interscapular brown adipose tissue (BAT), a site of nonshivering thermogenesis, we employed three lines of transgenic mice. Two of the lines are dwarf due to expression of a GH antagonist (GHA) or disruption of the GH receptor/binding-protein gene. A third mouse line is giant due to overexpression of a bovine GH (bGH) transgene. We have found that the body weights of those animals are proportional to their body lengths at 10 weeks of age. However, GHA dwarf mice tend to catch up with the nontransgenic (NT) littermates in body weight but not in body length at 52 weeks of age. The increase of body mass index (BMI) for GHA mice accelerates rapidly relative to controls as a function of age. We have also observed that BAT in both dwarf mouse lines but not in giant mice is enlarged in contrast to nontransgenic littermates. This enlargement occurs as a function of age. Northern analysis suggests that BAT can be a GH-responsive tissue because GHR/BP mRNAs were found there. Finally, the level of uncoupling protein-1 (UCP1) RNA was found to be higher in dwarf mice and lower in giant animals relative to controls, suggesting that GH-mediated signaling may negatively regulate UCP1 gene expression in BAT.     

Studies on tilapia hepatic growth hormone receptor.

Tilapia liver membranes were solubilized with 1% Triton X-100. The presence of growth hormone (GH) receptors was demonstrated by specific binding of radioiodinated tilapia GH (125I-tGH). The solubilized receptor possessed a molecular weight of around 400,000. It was adsorbed on Con A-Sepharose and DEAE BioGel A indicating that it contains carbohydrates and is acidic in character. Its protein nature was revealed by destruction of GH-binding activity by proteases. The involvement of essential sulfhydryl group was suggested by inhibition of 125I-tGH binding to the solubilized receptor by p-chloromercuribenzene sulfonate which could be reversed by dithioerythritol treatment.     

Influence of triiodothyronine and growth hormone on growth of dwarf and normal chickens: interactions of hormones and genotype

The effects of dietary triiodothyronine (T3), injections of a preparation of growth hormone (GH) (purified from chicken pituitary tissue) and their combination on growth were investigated in three lines of chickens. The three lines were the Cornell K strain (K) (a single Comb White Leghorn strain), the Cornell K strain hemizygous for the sex-linked dwarfing gene (SLD), and the Cornell K strain homozygous recessive for the autosomal dwarfing gene (ADW). A dietary T3 treatment by genotype interaction was observed. Dietary T3 (0.1 ppm) decreased growth in the K line, tended to decrease growth in the ADW line while it tended to increase growth in the SLD line. Chicken growth hormone (100 micrograms/kg body wt) alone did not affect growth in any of the lines studied. There was, however, a GH treatment by T3 treatment interaction. Chicken GH overcame the growth-depressing effects of T3 in the K and ADW lines while it tended to promote growth in T3 treated SLD birds. Dwarf (SLD) chickens had higher basal circulating GH concentrations, lower circulating immunoreactive somatomedin C concentrations, and lower circulating T3 concentrations than the K or ADW chickens.     

Chronic intravenous infusion of chicken growth hormone increases body fat content of young broiler chickens

The purpose of this study was to determine the effects of programmed intravenous infusion of chicken growth hormone (cGH) on growth and metabolism of young broiler chickens (4–7 weeks of age). Four-week-old broiler cockerels, fitted with indwelling jugular catherters, were randomly assigned to three treatment groups (6 birds/group): pulsatile infusion of buffer (phosphate buffer, pH 7.4)[PB-P] at 3 hr intervals, pulsatile infusion of cGH (15 μg/kg at 3 hr intervals)[GH-P], or continuous infusion of cGH (120 μg/kg-day)[GH-C]. Birds were bled 5 min before (0-min) and 5 min post-infusion (relative to the pulses of PB and cGH) at 5, 6, and 7 weeks of age. Pulsatile infusion of cGH increased (P < 0.05) feed consumption by 24% and reduced (P < 0.05) feed efficiency by 14% without affecting body weight (BW) gain. The relative weights (%BW) of liver, abdominal fat, and bursa of Fabricius were not affected by the pattern of cGH infusion. However, the body fat content of cGH-infused chickens was increased (P < 0.05) by 13% (GH-C) and 17% (GH-P), while body protein and water contents were slightly reduced. Body ash content was not affected by pattern of cGH infusion. When compared with the PB-P controls, the GH-P treatment depressed (P < 0.05) hepatic GH-binding activity by 52% without affecting plasma insulin-like growth factor-I (IGF-I) levels. Continuous infusion of cGH increased (P < 0.05) plasma IGF-I by 16%, thyroxine (T₄) by 31%, and glucagon levels by 55%, although plasma GH levels were only 47% higher than those of the PB-P group. However, the GH-P treatment was only half as effective as the GH-C pattern in elevating plasma levels of T₄ and glucagon. This study shows that programmed intravenous infusion of cGH increases deposition of body fat in young rapidly-growing broiler chickens.     

Stimulation of growth hormone release in dwarf and normal chickens by thyrotrophin releasing hormone (TRH) or human pancreatic growth hormone releasing factor (hpGRF)

The effect of thyrotrophin releasing hormone (TRH) or human pancreatic growth hormone releasing factor (hpGRF) on growth hormone (GH) release was studied in both dwarf and normal Rhode Island Red chickens with a similar genotype except for a sex-linked dw gene. Both TRH (10 micrograms/kg) and hpGRF (20 micrograms/kg) injections stimulated plasma GH release within 15 min in young and adult chickens. The increase in GH release was higher in young cockerels than that in adult chickens. The age-related decline in the response to TRH stimulation was observed in both strains, while hpGRF was a still potent GH-releaser in adult chickens. The maximal and long acting response was observed in young dwarf chickens, suggesting differences in GH pools releasable by TRH and GRF in the anterior pituitary gland. The pituitary gland was stimulated directly by perifusion with hpGRF (1 microgram/ml and 10 micrograms/ml) or TRH (1 microgram/ml). Repeated perifusion of GRF at 40 min intervals blunted further increase in GH release, but successive perifusion with TRH stimulated GH release. The results suggest the possibility that desensitization to the effects of hpGRF occurs in vitro and that the extent of response depends on the number of receptors for hpGRF or TRH and/or the amount of GH stored in the pituitary gland.     

Dietary thyrotropin-releasing hormone stimulates growth rate and increases the insulin: glucagon molar ratio of broiler chickens

The effects of thyroid manipulation on growth, feed efficiency, and plasma hormone levels were determined in rapidly growing chickens. Beginning at 3 weeks of age, eight broiler cockerels were provided with control feed (CF) or feed containing either 1 ppm of triiodothyronine (T3), 1 ppm of thyroxine (T4), 0.3% propylthiouracil (PTU), or 5 ppm of thyrotropin-releasing hormone (TRH) for 3 weeks. Blood samples were taken at 4, 5, and 6 weeks for determination of plasma levels of growth hormone, insulin-like growth factor, T3, T4, insulin, glucagon, glucose, and nonesterified fatty acids. Dietary TRH increased (P less than 0.05) the growth rate of chickens by 14% when compared with the CF group. Plasma growth hormone levels were reduced (P less than 0.05) 65% by dietary T3 and 33% by treatment with either T4 or TRH when compared with the CF group. Plasma insulin-like growth factor levels were 16% lower (P less than 0.05) in PTU-fed birds than the other treatment groups. Plasma T3 levels were elevated (P less than 0.05) 3-fold by dietary T3 and 38% by TRH whereas plasma T3 in the PTU group was 38% below the average of CF birds. Plasma T4 levels were increased (P less than 0.05) by 12-fold in T4-fed birds, decreased 48% in TRH-fed birds, and nondetectable in birds treated with either T3 or PTU. Compared with the other treatments, dietary PTU increased (P less than 0.01) plasma insulin levels 4.3-fold whereas TRH provided a 2.7-fold increase in plasma insulin. Plasma glucagon levels were 26% higher (P less than 0.05) in T3-fed birds than those fed either T4 or PTU. These observations indicate that thyroid activity plays an important role in regulating secretion of GH and the pancreatic hormones. Furthermore, our study demonstrates the potential use of TRH as an orally active growth promoter for poultry.     

Tyrosine kinases compete for growth hormone receptor binding and regulate receptor mobility and degradation

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Effect of age and growth rate on myocardial irritability in broiler chickens

Young broiler chickens are undergoing a period of extremely rapid growth and may be expected to be in a state of extreme endocrine and biochemical flux. These birds are also subject to a sudden death syndrome of unknown etiology. We hypothesized that an increased myocardial sensitivity in birds exhibiting early rapid growth may contribute to this syndrome. The objective of the current study was to investigate the interaction between early growth rate and age on myocardial irritability in young broiler chickens. This study utilized 74 male broiler chickens between 3 (Group A, 21-24 days) and 6 (Group B, 43-47 days) weeks of age exhibiting rapid (heavy weight, greater than 425 g at 2 weeks) and slow (low weight, less than 350 g at 2 weeks) early growth. Physiologic parameters such as blood pressure, heart rate, and arterial blood gases in the awake, restrained birds were essentially unchanged across these groups. Myocardial irritability of the anesthetized bird (pentobarbital) as measured by the threshold to electrical fibrillation was significantly increased only in the Group A HW birds. The results of this study suggest an increased myocardial irritability in large young broilers (3 weeks of age) that is no longer present in a similar group of older birds (6 weeks of age). These findings are consistent with the hypothesis that there is an increased myocardial sensitivity in birds succumbing to sudden death syndrome, with death due to myocardial fibrillation. Further research is needed to determine the mechanisms involve.     

Comparative proteomics and phosphoproteomics analyses of DHEA-induced on hepatic lipid metabolism in broiler chickens

Dehydroepiandrosterone (DHEA) is a precursor of the adrenocorticosteroid hormones that are common to all animals, including poultry. The study described herein was undertaken to investigate the effect of DHEA on lipid metabolism in broiler chickens during embryonic development and to determine the regulatory mechanisms involved in its physiological action. Treatment group eggs were injected with 50 mg DHEA diluted in 50 μL dimethyl sulfoxide (DMSO) per kg, while control group eggs (arbor acres [AA] fertilized) were injected with 50 μL DMSO per kg prior to incubation. Liver samples were collected on days 9, 14 and 19 of embryonic development as well as at hatching. Extracted proteins were analyzed by two dimensional gel electrophoresis (2-DE) in combination with western blotting for specific anti-phosphotyrosine. The differential spots were identified by matrix-assisted laser desorption/ionization-time of flight mass spectrometry (MALDI-TOF-MS) or MALDI-TOF-TOF-MS. Peptide mass fingerprinting (PMF) of the differentially-expressed proteins were performed using the MASCOT, Prospector or proFound server. Thirty-seven proteins and twenty-two tyrosine phosphorylation proteins were successfully identified. All 37 proteins and 22 tyrosine phosphorylation proteins exhibited a significant volume difference from the control group. These results demonstrated that DHEA increased the expression and level of tyrosine phosphorylation and sulfotransferase proteins in broilers (at pI 5.9), therefore promoting the biotransformation of DHEA. The expression of apolipoproteinA-I was increased in the DHEA treatment group, which facilitated the conversion of cholesterol to cholesterol esters. Also, DHEA increased the expression of peroxiredoxin-6 and its tyrosine phosphorylation protein levels, thus enhancing its anti-oxidative activity. Furthermore, pyruvate dehydrogenase expression was decreased and the level of its tyrosine phosphorylation proteins increased in the DHEA treatment group. Take together, those data indicate that DHEA reduces the supply of acetyl-CoA by inhibiting the activity of its target enzyme (i.e. pyruvate dehydrogenase), thus affecting both protein synthesis and phosphorylation level and decreasing fat deposition in broiler chickens during embryonic development, which could reflect a physiologically-relevant DHEA fat-reduction mechanism in the broiler chicken.     

Organization of the sex-linked late-feathering haplotype in chickens

Nucleotide sequence analysis of polymerase chain reaction products confirmed that ev 21 integrated into one of two large homologous elements on the Z chromosome of late-feathering (LF) White Leghorn chickens. Southern blots of Not I-, Nae I-, Ksp I- and Bam HI-digested DNA from early-feathering (EF) and LF White Leghorns, that had been hybridized with a probe that flanks ev 21, indicated a 180 kb duplication of an unoccupied repeat in the LF genotype of White Leghorns. A Ksp I fragment that carries ev 21 was about 32 kb smaller than the Ksp I fragment found in EF DNA. In the evolution of LF, retroviral insertion into one of two large repeats and a 32 kb deletion may have generated LF.