Loss of sensitivity to insulin at early events of the insulin signaling pathway in the liver of growth hormone-transgenic mice.
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Growth hormone (GH) excess is associated with secondary hyperinsulinemia, but the molecular mechanism and consequences of this alteration are poorly understood. To address this problem we have examined the levels and phosphorylation state of the insulin receptor (IR) and the insulin receptor substrate-1 (IRS-1), the association between IRS-1 and the p85 subunit of phosphatidylinositol 3-kinase (PI 3-kinase) as well as the PI 3-kinase activity in the livers of GH-transgenic mice. As expected, IR levels were reduced in the liver of GH-transgenic mice (55% of normal values) as determined by immunoblotting with an anti-IR beta-subunit antibody. IR and IRS-1 phosphorylation as determined by immunoblotting with antiphosphotyrosine antibody were increased in basal conditions by 315% and 560% respectively. After a bolus administration of insulin in vivo, IR phosphorylation increased by 40% while IRS-1 phosphorylation did not change. Insulin administration to control (normal) mice produced 670% and 300% increases in the IR and IRS-1 phosphorylation respectively. In the GH-transgenic animals, basal association of PI 3-kinase with IRS-1 as well as PI 3-kinase activity in liver was increased by 200% and 280% respectively, and did not increase further after administration of insulin in vivo, indicating a complete insensitivity to insulin at these levels. In conclusion, GH excess and the resulting secondary hyperinsulinemia were associated with alterations at the early steps of insulin action in liver. IR concentration was reduced, while IR and IRS-1 phosphorylation, IRS-1/PI 3-kinase association, and PI 3-kinase activity appeared to be maximally activated under basal conditions, thus making this tissue insensitive to further stimulation by exogenous insulin in vivo. (+info)
Effects of GH, prolactin and cortisol on hepatic heat shock protein 70 expression in a marine teleost Sparus sarba.
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Heat shock protein 70 (HSP70) expression was assessed in hepatic tissue of a marine teleost Sparus sarba after exogenous hormone administration. Using a PCR-amplified, homologous HSP70 cDNA clone, as a probe in Northern analysis, we detected a 2.3 kb transcript which was elevated after exposure to a temperature 7 degrees C above the ambient. For our studies on hormonal effects on HSP70 expression, groups of fish were administered recombinant bream GH (rbGH), ovine prolactin (oPRL) or cortisol daily over a 7-day period. Quantification of hepatic HSP70 transcript revealed that the administration of GH and PRL significantly reduced HSP70 mRNA abundance by 42% and 54% from saline-injected fish respectively. Also hepatic HSP70 levels were reduced by 76% and 64% as determined by immunoblotting after rbGH and oPRL treatment respectively. The administration of exogenous cortisol did not alter hepatic HSP70 mRNA or protein levels in S. sarba. The results obtained in this study are the first evidence for hormonal modulation of heat shock protein expression in fish. The significance of these results is discussed within the context of current knowledge on the roles of these hormones in teleostean stress response. (+info)
A missense mutation in the GHR gene of Cornell sex-linked dwarf chickens does not abolish serum GH binding.
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Sex-linked dwarfism (SLD) in chickens is characterized by impaired growth despite normal or supranormal plasma growth hormone (GH) levels. This resistance to GH action is thought to be due to mutations of the GH receptor (GHR) gene that reduce or prevent GH binding to target sites. The genetic lesion causing GH resistance in Cornell SLD chickens is, however, not known. Previous studies have shown that hepatic GH-binding activity is abnormally low in these birds, yet the GHR gene is transcribed into a transcript of appropriate size and abundance. Point mutations or defects in translation could therefore account for the impaired GHR activity in this strain. These possibilities were addressed in the present study. A missense mutation resulting in the substitution of serine for the conserved phenylalanine was identified in the region of the GHR cDNA encoding the extracellular domain. Translation of this mutant transcript was indicated by the presence of GHR/GH-binding protein (GHBP)-immunoreactive proteins in liver (55, 70 and 100 kDa) and serum (70 kDa) of normal (K) and SLD birds. Radiolabelled GH did not, however, bind to the hepatic membranes of most SLD chickens. Serum GH-binding activity, in contrast, was readily detectable, although at significantly lower levels than in normal birds. The missense mutation in the SLD GHR gene may thus affect targeting of GHRs to hepatic plasma membranes. (+info)
Regulation of differentiation of sheep subcutaneous and abdominal preadipocytes in culture.
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Factors regulating the differentiation of sheep subcutaneous and abdominal (omental or perirenal) preadipocytes from foetal lambs, suckling lambs and fattening sheep have been investigated using a serum-free cell culture system. Differentiation was assessed by changes in the activity of the enzyme glycerol 3-phosphate dehydrogenase. Insulin or IGF-I was essential for differentiation. Dexamethasone, a lipid supplement (Excyte) and the thiazolidinedione, rosiglitazone (BRL 49653) (a peroxisome proliferator-activated receptor-gamma (PPAR-gamma) agonist) all enhanced preadipocyte differentiation, whereas fibroblast growth factor and GH were inhibitory. The most effective combination was insulin, triiodothyronine, dexamethasone and rosiglitazone. Under suboptimal conditions, preadipocytes from fattening sheep differentiated less well than cells from suckling and foetal lambs. Also, under suboptimal conditions, abdominal preadipocytes did not differentiate as well as subcutaneous preadipocytes. However, age and depot differences were minimised when cells were cultured with insulin, triiodothyronine, rosiglitazone and either dexamethasone or lipid. The results suggest that variation in the ability to produce the natural ligand for PPAR-gamma contributes to depot- and age-specific differences in the ability of preadipocytes to differentiate. (+info)
Inhibitory effect of a growth hormone receptor antagonist (G120K-PEG) on renal enlargement, glomerular hypertrophy, and urinary albumin excretion in experimental diabetes in mice.
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Growth hormone (GH) and IGFs have a long and distinguished history in diabetes, with possible participation in the development of renal complications. To investigate the effect of a newly developed GH receptor (GHR) antagonist (G120K-PEG) on renal/glomerular hypertrophy and urinary albumin excretion (UAE), streptozotocin-induced diabetic and nondiabetic mice were injected with G120K-PEG every 2nd day for 28 days. Placebo-treated diabetic and nondiabetic animals were used as reference groups. Placebo-treated diabetic animals were characterized by growth retardation, hyperphagia, hyperglycemia, increased serum GH levels, reduced serum IGF-I, IGF-binding protein (IGFBP)-3, and liver IGF-I levels, increased kidney IGF-I, renal/glomerular hypertrophy, and increased UAE when compared with nondiabetic animals. No differences were seen between the two diabetic groups with respect to body weight, food intake, blood glucose, serum GH, IGF-I, and IGFBP-3 levels or hepatic IGF-I levels. Kidney IGF-I, kidney weight, and glomerular volume were normalized, while the rise in UAE was partially attenuated in the G120K-PEG-treated diabetic animals. No effect of G120K-PEG treatment on any of the parameters mentioned above was seen in nondiabetic animals. In conclusion, administration of a GHR antagonist in diabetic mice has renal effects without affecting metabolic control and circulating levels of GH, IGF-I, or IGFBP-3, thus indicating that the effect of G120K-PEG may be mediated through a direct inhibitory effect on renal IGF-I through the renal GHR. The present study suggests that specific GHR blockade may present a new concept in the treatment of diabetic kidney disease. (+info)
A pharmacokinetic/pharmacodynamic model for recombinant human growth hormone effects on induction of insulin-like growth factor I in monkeys.
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The pharmacokinetics of recombinant human growth hormone (rhGH) and its effects on the induction of insulin-like growth factor I (IGF-I) were studied in juvenile rhesus monkeys. Disposition profiles of rhGH from two short-term i.v. infusion studies were described by a two-compartment model yielding a clearance of 16.1 ml/min and T1/2 of 2.0 h. Four rhGH treatment groups were included in this study: group A, ProLease rhGH (24 mg), a sustained-release microsphere formulation; group B, a single s.c. injection plus an implanted osmotic pump (24.4 mg); group C, a single s.c. injection (25.9 mg); group D, daily 0.86-mg s.c. injection for 28 days. Their rhGH input profiles were analyzed by a numerical deconvolution method. ProLease and osmotic pump provided zero-order inputs of rhGH and maintained the serum rhGH concentrations around 9 to 13 ng/ml for 16 (group A) and 30 days (group B). For s.c. injections, rhGH underwent first-order absorption. An indirect response model was applied based on use of a Hill function for stimulation of IGF-I production. Parameter values obtained included Smax = 2.2, SC50 = 6.5 ng/ml, and gamma (slope coefficient) = 6.8, which were applicable to all treatments. The area under effect curve showed group B to be most effective for IGF-I induction, whereas group A produced the highest peak level in 16 days. Group C had the lowest induction among the four groups, despite being given the highest dose. Group D had modest IGF-I induction, but the pulsatile rhGH input is less effective than continuous input provided by ProLease. Our pharmacokinetic/pharmacodynamic model demonstrates that ProLease and osmotic pump delivery were best able to maintain rhGH level above the s.c.50 value, which provided more effective IGF-I induction compared with the single or daily subcutaneous injections in solution. (+info)
Effects of bovine somatotropin and ruminally undegraded protein on feed intake, live weight gain, and mohair production by yearling Angora wethers.
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Yearling Angora wethers (n = 24; 24+/-1.0 kg BW) were used in an experiment with a 2 x 2 factorial arrangement of treatments to investigate effects of bovine somatotropin (bST) treatment and dietary level of ruminally undegraded protein on DMI, ADG, and mohair production. Untreated casein (UC) or casein treated with formaldehyde (TC) was included at 7% DM of a diet containing 11% CP and 46% concentrate. A slow-release bST form was administered weekly to deliver 0 (Control) or 100 microg/ (kg BW.d) of bST. Plasma concentrations of bST and IGF-I were increased (P < .05) during the 7-d period following bST injection. Ruminal fluid ammonia N concentration was lower (P < .01) for TC than for UC before feeding (6.6 vs 7.5 mg/dL) and 4 h later (8.2 vs 12.2 mg/dL), and total VFA concentration was lower (P < .01) for TC than for UC. Treatment with bST decreased (P = .08) DMI with UC (1.15 vs .91 kg/d) and increased (P = .08) DMI with TC (.95 vs 1.06 kg/d). Formaldehyde treatment of casein increased ADG (65, 74, 55, and 91 g/d; P = .03) and clean fleece production (P < .01; 14.1, 17.3, 15.0, and 18.4 g/d for UC-Control, TC-Control, UC-bST, and TC-bST, respectively), with no effect of bST during the 8-wk period of treatment or for the 8 wk thereafter (P > .10). In conclusion, with yearling Angora wethers, bST does not seem useful to enhance mohair production and may not alter effects of dietary level of ruminally undegradable protein on mohair production. (+info)
Components of growth in mice hemizygous for a MT/bGH transgene.
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The objective of this study was to determine the effects of a metallothionein/bovine GH transgene on duration and rate of growth of lean and fat in mice. Mice were produced by mating hemizygous transgenic males to nontransgenic females. Ten weights and six measurements of total body electrical conductivity to estimate body composition were taken on 147 progeny between birth and 84 d of age. Growth traits for fat-free mass (FFM) and body fat mass (FM) were obtained by fitting FFM and FM to a logistic curve y = A/(1 + exp(k(b - t))), where y is FFM or FM, A is asymptotic mass, k and b are curve parameters, and t is time in days. The function and its first, second, and third derivatives for FFM and FM were used to model growth. A mixed model was used with animal and litter as random effects and trans-genotype, sex, and transgenotype x sex as fixed effects in analyses of growth traits. Estimates of transgeno-type and transgenotype x sex interaction were tested by using their corresponding standard errors. Males had greater response to the transgene than females in final FFM and growth rate during the entire growth period. Transgenic males and females had greater duration of lean growth than nontransgenics. Transgenic males began to accumulate fat later, but they eventually gained more fat than transgenic females. (+info)