Relationships between the thyroid and somatotropic axes in steers I: Effects of propylthiouracil-induced hypothyroidism on growth hormone, thyroid stimulating hormone and insulin-like growth factor I

The effects of propylthiouracil (PTU)-induced thyroid hormone imbalance on GH, TSH and IGF-I status in cattle were examined. In the first study, four crossbred steers (avg wt 350 kg) were fed a diet dressed with PTU (0, 1, 2 or 4 mg/kg/d BW) in a Latin square design with four 35-d periods. On day 29 in each period, steers were challenged with an intrajugular bolus of thyrotropin releasing hormone (TRH, 1.0 μg/kg). Blood samples were obtained to assess the change in plasma GH and TSH as affected by PTU. Plasma IGF-I was measured from blood samples obtained before and after (every 6 hr for 24 hr) intramuscular injection of bovine GH (0.1 mg/kg, day 31). Doses of 1 and 2 mg/kg PTU increased plasma T₄ (P<.01). At 4 mg/kg, PTU depressed T₄ concentrations to 30% of control (P<.01). Plasma T₃ linearly decreased with increasing doses of PTU (P<.01). Plasma TSH increased when PTU was fed at 4 mg/kg (P<.05) while the TSH response to TRH declined with increasing PTU (P<.02). Neither basal nor TRH-stimulated plasma concentration of GH was affected by PTU; the IGF-I response to GH tended to increase at the 1 and 2 mg/kg PTU (P<.01). In a second study 24 crossbred steers were fed PTU (1.5 mg/kg) for 119 d in a 2 × 2 factorial design with implantation of the steroid growth effector, Synovex-S (200 mg progesterone + 20 mg estradiol), as the other main effect. Basal plasma GH and IGF-I were not affected by PTU treatment. Synovex increased plasma concentration (P<.01) of IGF-I without an effect on plasma GH. The data suggest that mild changes in thyroid status associated with PTU affects regulation of T₃, T₄ and TSH more than GH or IGF-I in steers.     

Effects of winter nutrition and summer pasture or a feedlot diet on plasma Insulin-like Growth Factor I (IGF-I) and the relationship between circulating concentrations of IGF-I and thyroid hormones in steers

Effects of two winter nutritional levels (LOW, MOD) and two summer pastures (bahiagrass, BG; perennial peanut, PP) on plasma IGF-I, and the relationship between IGF-I and average daily gain (ADG), thyroid hormones, plasma urea, packed cell volume (PCV) and steer type were determined in 101 steers (217 kg) varying in breed composition, frame size and initial condition. Relationships between body composition or composition of gain and IGF-I were determined in 11 contemporary steers assigned directly to the feedlot. Initial IGF-I (57.9 ± 3.5 ng/ml) was positively correlated (P<.05) to initial condition, estimated percentage of Brahman and plasma T₃, but not related to subsequent ADG. During the 126-day wintering period, ADG was .21 kg for the LOW winter treatment and .47 kg for the MOD winter treatment. Concentration of IGF-I in the wintering period was affected (P<.01) by nutritional level (LOW = 71.8 ng/ml, MOD = 150.6 ng/ml) and was positively related to winter ADG in MOD steers (P<.01) but not in LOW steers. Concentration of IGF-I in winter was also positively related to condition at the end of the winter period (P<.01), T₃ (P<.05) and T₄ (P<.05). There were no effects of winter treatment on IGF-I during the subsequent summer pasture period. During the 145-d summer period, ADG was .53 kg for BG and .68 kg for PP. Concentration of IGF-I during the summer period was affected (P<.05) by pasture treatment (BG = 138.6 ng/ml, PP = 181.9 ng/ml), was positively related (P<.01) to PCV and percentage of Brahman, and was negatively related (P<.05) to estimated percentage of English breeding. In steers assigned directly to the feedlot, IGF-I was correlated with empty body (EB) weight (r=−.59, P<.10), EB water (r=−.59, P<.10) and EB protein (r=−.60, P<.10) at slaughter, and with days on feed (r=−.65, P<.05), but was not correlated with ADG or rate of component gain. These data indicate that IGF-I is related to nutritional status in steers as in other species, that there may be significant breed or cattle type differences in circulating concentrations of IGF-I, and that circulating concentration of IGF-I may be functionally related to plasma concentration of thyroid hormones.     

Perturbed metabolism and hormonal profiles in calves infected with Sarcocystis cruzi

Plasma concentrations of growth hormone (GH), thyroid stimulating hormone (TSH), insulin (IN), thyroxine (T₄), and triiodothyronine (T₃) in addition to metabolic parameters [N balance (NB), urinary 3-methylhistidine (TMH), urinary creatinine (CR), and urinary hydroxyproline (HP)] were measured in 4-mo-old Holstein steers divided equally among groups infected with Sarcocystis (I), noninfected ad libitum fed (C), and noninfected pair fed to I (PF) (7 steers per treatment). Effects of infection on these parameters beyond those attributable to altered dietary intake were determined using orthogonal contrasts (effect of intake, C vs I + PF; effect of infection, PF vs I). NB was higher in C than I and PF (P<.05) and lower in I than PF (P<.02). Hydroxyproline and CR were influenced by intake (P<.05) and HP excretion was reduced in association with infection (P<.05). Reduced intake was associated with lowered mean basal plasma concentrations of GH, IN, T₃ and T₄ (P<.05). Infection further reduced (P<.001) plasma T₃ concentration.

Triiodothyronine and T₄ responses following an intravenous bolus of thyrotropin releasing hormone (TRH) were measured. The magnitude of the responses in I and PF were lower than those observed in C (P<.05). Plasma T₃ responses were further reduced in association with infection (P<.05). Insulin responses to intravenous arginine infusion (ARG) were also low in association with reduced intake. Growth hormone responses to TRH or ARG were affected by the level of feed intake only. These data suggest that hormonal perturbations associated with the insult of infection further compromise metabolism and the direction of nutrient partitioning that would ordinarily be associated with developmental growth in young steers beyond those responses anticipated from solely the reduction of feed intake.     

Effects of cimaterol administration on plasma concentrations of various hormones and metabolites in friesian steers

The aim of the experiment was to determine the acute and chronic effects of the β-agonist, cimaterol, on plasma hormone and metabolite concentrations in steers. Twelve Friesian steers (liveweight = 488 ± 3 kg) were randomly assigned to receive either 0 (control; n=6) or .09 mg cimaterol/kg body weight/day (treated; n=6). Steers were fed grass silage ad libitum. Cimaterol, dissolved in 140 ml of acidified distilled water (pH 4.2), was administered orally at 1400 hr each d. After 13 d of treatment with cimaterol or vehicle (days 1 to 13), all animals were treated with vehicle for a further 7 d (days 14 to 20). On days 1, 13 and 20, blood samples were collected at 20 min-intervals for 4 hr before and 8 hr after cimaterol or vehicle dosing. All samples were assayed for growth hormone (GH) and insulin, while samples taken at −4, −2, 0, +2, +4, +6 and +8 hr relative to dosing were assayed for thyroxine (T₄), triiodothyronine (T₃), cortisol, urea, glucose and non-esterified fatty acids (NEFA). Samples taken at −3 and +3 hr relative to dosing were assayed for IGF-I only. On day 1, cimaterol acutely reduced (P<.05) GH and urea concentrations (7.6 vs 2.9 ± 1.4 ng/ml; and 6.0 vs 4.9 ± 0.45 mmol/l, respectively; mean control vs mean treated ± pooled standard error of difference), and increased (P<.05) NEFA, glucose and insulin concentrations (160 vs 276 ± 22 μmol/l, 4.1 vs 6.2 ± 0.15 mmol/l and 29.9 vs 179.7 ± 13.9 μU/ml, respectively). Plasma IGF-I, T₃, T₄ and cortisol concentrations were not altered by treatment. On day 13, cimaterol increased (P<.05) GH and NEFA concentrations (7.7 vs 14.5 ± 1.4 ng/ml and 202 vs 310 ± 22 mEq/l, respectively) and reduced (P<.05) plasma IGF-I concentrations (1296 vs 776 ± 227 ng/ml). Seven-d withdrawal of cimaterol (day 20) returned hormone and metabolite concentrations to control values. It is concluded that : 1) cimaterol acutely increased insulin, glucose and NEFA and decreased GH and urea concentrations, 2) cimaterol chronically increased GH and NEFA and decreased IGF-I concentrations, and 3) there was no residual effect of cimaterol following a 7-d withdrawal period.     

Effect of daily injections of growth hormone-releasing factor and thyrotropin-releasing hormone on growth and endocrine parameters in chickens

The control of growth is a complex mechanism regulated by several metabolic hormones including growth hormone (GH) and thyroid hormones. In avian species, as well as in mammals, GH secretion is regulated by hypothalamic hypophysiotropic hormones. Since thyrotropin-releasing hormone (TRH) and growth hormone-releasing factor (GRF) are potent GH secretagogues in poultry, we were interested in determining the influence of daily intravenous administration of either peptide or both simultaneously on circulating GH and IGF-I concentrations and whether an improvement in growth rate or efficiency would be obtained.

Male broiler chicks were injected once daily for a period of 21 days with either GRF (10 μg/kg), TRH (1 μg/kg) or both GRF and TRH (10 and 1 μg/kg respectively) between four and seven weeks of age. On the last day of the experiment, following intravenous injection of TRH, GRF or a combination of GRF and TRH, plasma GH levels were significantly (P<.05) increased to a similar extent in control chicks and in those which had received daily peptide injections for the previous 21 days. Circulating GH levels between 10 and 90 min post-injection were significantly (P<.05) greater and more than additive than GH levels in chicks injected with both GRF and TRH when compared to those injected with either peptide alone. Mean plasma T₃ concentrations during that same time period were significantly elevated (P<.05) above saline-injected control chick levels in birds treated with TRH or GRF and TRH respectively, regardless of whether the chicks had received peptide injections for the previous 21 days. There was no evidence of pituitary refractoriness to chronic administration of either TRH or GRF injection in terms of growth or thyroid hormone secretion.

Despite the large elevation in GH concentration each day, growth rate, feed efficiency and circulating IGF-I concentrations were not enhanced. Thus the quantity or secretory pattern of GH secretion induced by TRH or GRF administration was not sufficient to increase plasma IGF-I concentration or growth.     

Effects of dietary T3 on growth parameters and hormone levels in normal and sex-linked dwarf chickens

Tri-iodothyronine (T₃) has been administered in the diet, from day of hatch until 8 weeks of age, to sex-linked dwarf and normal chicks of both sexes from a brown-egg slow-growing strain. Feed was supplemented with either 0.1 ppm or 0.5 ppm T₃. A significant genotype by treatment interaction was observed on body weight: the effect of T₃ in males was significantly positive for dwarfs and null for normals, the effect in females was null for dwarfs and significantly negative for normals. Feed efficiency was rather decreased by the treatment in both genotypes. Abdominal fatness was decreased in a dose-dependent manner in both genotypes, while rectal temperature was raised in dwarf chicks only. Plasma T₃ was raised to normal levels in dwarfs receiving 0.1 ppm exogenous T₃, while the 0.5 ppm dose yielded hyperthyroid levels. Plasma GH levels were decreased in a dose-dependent manner by the T₃ treatment, the effect being larger in dwarfs. Surprisingly, plasma IGF-I was unchanged in spite of the GH decrease, whatever the genotype or the sex. It was concluded that exogenous T₃ alone can have a stimulatory effect on growth in dwarfs but can not fully restore a normal growth rate. Both T₃ and IGF-I are important for a normal growth and the relationships between T₃ and IGF-I production should be further investigated in order to better understand the physiological modifications due to the sex-linked dwarf gene.     

Response of young broiler chickens to chronic injection of recombinant-derived human insulin-like growth factor-I

The purpose of this study was to determine if exogenous insulin-like growth factor-I (IGF-I) would improve growth rate or body composition of young broiler chickens. Broiler cockerels were given a daily intramuscular (im) injection of sodium acetate buffer (buffer control), 100 or 200 μg recombinant-derived human IGF-I (rhIGF-I) per kg body weight from 11 to 24 days of age. Exogenous IGF-I did not affect the average daily gain, average daily feed consumption, or the gain-to-feed ratio of broiler chickens. Although daily injection of 200 μg/kg of rhIGF-I reduced (P<0.05) body ash content, there was no significant effect of IGF-I treatment on either body fat or protein content. Plasma GH levels were depressed (P<0.05) by chronic treatment with rhIGF-I. In contrast, plasma levels of T₃ and T₄ were not affected by rhIGF-I treatment. The half-life of rhIGF-I in plasma was determined at 25 days of age in naive control or chronically-injected chickens after a single intravenous dose of 50 μg rhIGF-I/kg. We found a single compartment, first-order disappearance pattern of rhIGF-I from chicken plasma. The half-life (t_1/2) of rhIGF-I in plasma was similar (t_1/2 = 32.5 min) for naive controls (injected once) or chronically-treated chickens which had received a daily injection of rhIGF-I (100 or 200 μg/kg) for 14 d. These data indicate that daily injection of IGF-I cannot be used to enhance growth performance or body composition of broiler chickens when given during the early growth period. The depression of plasma GH levels in rhIGF-I-injected chickens supports a negative-feedback role of IGF-I on pituitary GH secretion.     

Effect of progressive cachectic parasitism and growth hormone treatment on hepatic 5'-deiodinase activity in calves

Thyroid status is compromised in a variety of acute and chronic infections. Conversion of thyroxine (T₄) into the metabolically active hormone, triiodothyronine (T₃), is catalyzed by 5′-deiodinase (5′D) mainly in extrathyroidal tissues. The objective of this study was to examine the effect of protozoan parasitic infection (Sarcocystis cruzi) on hepatic 5′D (type I) activity and plasma concentrations of T₃ and T₄ in placebo- or bovine GH (bGH)-injected calves. Holstein bull calves (127.5±2.0 kg BW) were assigned to control (C, ad libitum fed), infected (I, 250,000 S. cruzi sporocysts per os, ad libitum fed), and pair-fed (PF, non-infected, fed to intake of I treatment) groups placebo-injected, and three similar groups injected daily with pituitary-derived bGH (USDA-B-1, 0.1 mg/kg, i.m.) designated as C_GH, I_GH and PF_GH. GH injections were initiated on day 20 post-infection (PI), 3–4 days prior to the onset of clinical signs of the acute phase response (APR), and were continued to day 56 PI at which time calves were euthanized for liver collection. Blood samples were collected on day 0, 28, and 55 PI. Alterations in nutritional intake did not affect type I 5′D in liver. Treatment with bGH increased (P<0.05) 5′D activity in C (24.6%) and PF (25.5%) but not in I calves. Compared to PF calves, infection with S. cruzi reduced 5′D activity 25% (P<0.05) and 47.8% (P<0.01) in placebo- and bGH-injected calves, respectively. Neither nutrition nor bGH treatment significantly affected plasma concentrations of T₄ and T₃ on day 28 and 55 PI. However, plasma thyroid hormones were reduced by infection. On day 28 PI, the average plasma concentrations of T₃ and T₄ were reduced in infected calves (I and I_GH) 36.4% (P<0.01) and 29.4% (P<0.05), respectively, compared to pair-fed calves (PF and PF_GH). On day 55 PI, plasma T₃ still remained lower (23.7%, P<0.01 versus PF) in infected calves while plasma T₄ returned to control values. The data suggest that parasitic infection in growing calves inhibits both thyroidal secretion and extrathyroidal T₄ to T₃ conversion during the APR. After recovery from the APR, thyroidal secretion returns to normal but basal and bGH-stimulated generation of T₃ in liver remains impaired.     

Secretory patterns of growth hormone and insulin-like growth factor-I during peripubertal period in intact and castrate male cattle

Fifteen Angus bulls and 15 Angus steers 9 months of age and 275 kg of body weight were bled at 20-min intervals over a 6-hr period and serum GH and IGF-I concentrations were measured by RIA. There were no differences between bulls and steers in the mean GH concentration, pulse frequency and amplitude when analyzed by the computer program PULSAR. Mean IGF-I concentration was not different between the two sex phenotypes, nor was there a significant correlation between the integrated IGF-I and GH concentrations. Subsequently, five bulls and five steers were selected from the 30 animals, full-fed a diet for growth in individual pens for 3 months and bled at 15-min intervals over a 24-hr period. Bulls tended to show a greater weight gain and feed conversion efficiency (P<.10) than steers during the 3-month period. Serum GH concentrations had a pulsatile pattern in all animals with no apparent diurnal rhythm during the 24-hr bleeding. Although mean GH concentration was not different between the two sex phenotypes, bulls tended to have lower baseline levels (P<.10) and greater peak amplitudes than steers. Serum IGF-I concentrations fluctuated within a two-fold concentration range, with no obvious pulsatility similar to that of GH. Mean IGF-I concentrations of each of the 10 animals were correlated with mean peak GH amplitudes (r = .79), but not with mean GH. These results suggest that gonadal hormone(s) modulates the GH secretory pattern and increases IGF-I secretion which may be related to the greater growth rate of bulls compared with steers.     

Effect of exogenous thyrotropin-releasing hormone (TRH) administration on plasma levels of triiodothyronine (T3), thyroxine (T4) and growth hormone (GH) in chronically catheterized suckling piglets.

The purpose of the present study was to determine experimental conditions to stimulate secretion of thyroid hormones (T₃ and T₄) with thyrotropin-releasing hormone (TRH) injections in suckling piglets during the first weeks of postnatal life. Three consecutive experiments were conducted. Four 10–20 d old piglets were i.m. injected with 0, 20, 100, 500 μg (experiment 1) or 0, 4, 20, 100 μg TRH/kg BW (experiment 2) according to a 4 × 4 latin square design involving different litters in each experiment. Blood samples were taken −15, −1, 15, 30, 45, 60, 90, 120 180 and 300 min after TRH injection in experiment 1, and −.25, −.08, .25, .5, 1, 2, 4, 6, 8, 12, 24, 30, 36, 48, 60 and 72 hr after TRH injection in experiment 2. T₃ and T₄ levels were significantly (P<.01) increased as soon as 30 and 45 min after TRH injection, respectively. Maximal levels of T₃ and T₄ were obtained 2 and 4 hr after the injection of 100 μg TRH. T₃ and T₄ returned to basal levels within 6 and 8 hr post injection, respectively. Plasma pGH levels were significantly (P<.001) increased 15 min after TRH injection in piglets injected with 500 μg. In experiment 3, 100 μg TRH/kg BW were injected i.m. either daily or every other day from .0 to 23 days of age. Results showed that T₄ response to TRH did not decrease after repeated injections. These results indicate that daily i.m. injections of 100 μg TRH/kg BW can be used to increase thyroid hormone levels for at least 13 d in the young suckling piglet.     

Influence of diet on basal and growth hormone-stimulated plasma concentrations of IGF-I in beef cattle

The effects of nutrition on plasma concentrations of insulin-like growth factor-I (IGF-I) were characterized in steers under basal conditions and following single i.m. injection of bovine growth hormone (bGH, .1 mg/kg BW). Nutritional effects on IGF-I were studied in three trials. In all trials steers were individually fed and penned Angus or Hereford x Angus (280 kg). In the first trial, two diets (LPLE1: 8% CP and 1.96 Mcal ME/kg, 4.5 kg.hd-1.d-1; MPHE1: 11% CP, 2.67 Mcal ME/kg, 6.5 kg.hd-1.d-1) were fed (n = 5/diet). Plasma IGF-I concentrations averaged 74 (LPLE1) and 152 (MPHE1) ng/ml (P less than .02). Following bGH injection, IGF-I increased to peak concentrations between 12 and 24 h (averaging 105 and 208 ng/ml at peak for LPLE and MPLE, respectively, P less than .01). In the second trial, steers were fed diets composed of 8, 11 or 14% CP and 1.96 or 2.67 Mcal ME/kg dry matter (6.35 kg.hd-1.d-1 in a factorial arrangement for 84 d, n = 4/diet). Within the low ME diet groups, plasma IGF-I was similar in steers fed 11 and 14% CP but greater at these two CP levels than in steers fed 8% CP (P less than .05). Within the high ME diet groups, plasma IGF-I increased linearly with CP (P less than .01). In the third trial, steers were fed diets to result in a negative N status. Insulin-like growth factor-I was lower (P less than .02) during feed restriction than when steers were full-fed. The IGF-I response to bGH was diminished or absent in underfed steers (P less than .01). These data are interpreted to suggest that diet composition and intake affect plasma concentrations of IGF-I in steers. In cattle, CP may be the primary nutritional determinant of basal IGF-I, but the IGF-I response to CP may be affected by the available ME. Undernutrition can attenuate the IGF-I response to GH and uncouple the regulation of IGF-I normally ascribed to GH.     

Extrathyroidal thyroxine-5′-monodeiodinase activity in cattle

The monodeiodination of thyroxine (T₄) to triiodothyronine (T₃) was studied in vitro using liver, kidney, and muscle obtained from two-year old Angus and Hereford steers. Tissues were homogenized in .1 M phosphate buffer-.25 M sucrose - 5 mM EDTA, pH 7.5, and centrifuged at 2000 × g for 30 min. Supernatants were incubated with T4 (1.3 μM) at 37 C and T₃ generated was measured by radioimmunoassay of an ethanol extract of the incubation mixture. The T₄ to T₃ conversion in Angus liver homogenate was dependent upon pH, temperature, duration of incubation (5–120 min), homogenate (.025–.20 g-eq tissue/ml), and substrate concentration (.32–6.43 μM T₄). The apparent K_m and V_max of the conversion were .64 μM T₄ and 1.87 ng T₃ generated/hr/mg protein, respectively. Mean T₄ to T₃ conversion in Angus liver and kidney was 1.37 and .22 ng T₃/hr/mg protein. The presence of 2 mM dithiothreitol (DTT), a sulfhydryl protective agent, significantly increased T₃ generation in liver and kidney (5.12 and 4.58 ng/hr/mg protein) and also revealed activity in muscle (05 ng/hr/mg protein). In liver and kidney from Hereford steers conversion activity was 2.89 and .48 in absence and 10.91 and 5.38 ng T₃/hr/mg protein in presence of DTT, respectively. These results demonstrate the presence of a very active enzymatic system responsible for the peripheral 5′-monodeiodination of T₄ to T₃ in cattle.     

Dietary fatty acids modulate hormone repsonses in lactating cows: Mechanistic role for 5′-deiodinase activity in tissue

Supplemental dietary fat provides excess fatty acids (FA), which can alter circulating concentrations of several hormones. To test the effects of fatty acid isomer type and possible sites of regulation, we abomasally infused fat mixtures high in cis-C_18:1 FA (iTRS) or no infusion (NI) and performed intravenous arginine (ARG) and intramuscular thyrotropin-releasing hormone (TRH) challenges. The experimental design was a replicated 3 × 3 Latin square. Challenges were conducted on Days 10 (ARG) and 12 (TRH) after initiation of fat infusion on each of three 4-wk experimental periods. Plasma concentrations of IGF-I were lower (P < 0.01) when cows received iCIS or iTRS compared with NI. Plasma insulin concentrations increased with ARG but responses were not affected by FA. Plasma growth hormone (GH) was unchanged after ARG. Peak plasma GH and thyroid-stimulating hormone (TSH) responses to TRH were blunted (P < 0.05 and P < 0.1, respectively), whereas thyroxine (T₄) and triiodothyronine (T3) responses were augmented post-TRH (P < 0.01) when cows received either FA isomer. Prolactin responses to TRH were not different between infusion treatments, although basal plasma concentrations before TRH were higher in cows infused with iTRS (P < 0.05). To focus on fat regulation of the thyroid axis, we tested directly in vitro the ability of fatty acids dissolved with sodium taurocholate to affect Type-I 5'-deiodinase (5'D) activity in bovine liver homogenates. Homogenate 5'D was not affected by C_2:0---C10:0 fatty acids, but decreased linearly (P < 0.01) with increasing concentrations of C_12:0---C_16:0 and C_18:1 isomers. CisC_18:1 decreased 5′D more than the trans-isomer (P < 0.01), the difference was only apparent at concentrations greater than 0.25 mM. The data suggest that various aspects of pituitary hormone regulation are differentially affected by FA composition. Fatty acid infusion may accentuate end organ responses in the thyroid axis and decrease IGF-I in the somatotropic axis. The data also suggest that FA isomer may alter patterns of extrathyroidal generation of thyroid hormones via direct influences on 5′D.     

Time course of changes in growth factor mRNA levels in muscle of steroid-implanted and nonimplanted steers

We used a muscle biopsy technique in conjunction with real-time PCR analysis to examine the time course of changes in muscle IGF-I, IGFBP-3, myostatin, and hepatocyte growth factor (HGF) mRNA in the longissimus muscles of Revalor-S-implanted and nonimplanted steers on d 0, 7, 12, and 26 after implantation (nine steers/treatment group). Administration of a Revalor-S implant increased (P < 0.01) ADG and improved (P < 0.05) feed efficiency, 36 and 34%, respectively, compared with steers that received no implant during the 26-d trial. Daily dry matter intake did not differ (P > 0.15) between nonimplanted and implanted steers. Steers receiving the Revalor-S implant had increased (P < 0.001) circulating IGF-I concentrations compared with nonimplanted steers. The longissimus muscles of steers receiving the Revalor-S implant contained increased (P < 0.001) IGF-I mRNA levels compared with longissimus muscles of nonimplanted steers over the 26-d duration of the study. Longissimus muscle IGF-I mRNA levels in implanted steers were increased (P < 0.003) relative to d-0 concentrations on d 7 and 12 (101% and 128%, respectively), and byd 26, longissimus muscle mRNA levels were more than three times (P < 0.0001) those in the longissimus muscles of the same steers on d 0. There was no treatment effect on the level of IGFBP-3, myostatin, or HGF mRNA in the longissimus muscle at any time point; however, levels of IGFBP-3, myostatin, and HGF mRNA increased with time on feed. Based on current and previous studies, we hypothesize that the increased IGF-I level in muscle of implanted steers by d 7 of implantation stimulates satellite cell proliferation and maintains a high number of proliferating satellite cells at a point in the growth curve where satellite cell numbers and activity are normally dropping off. This would prolong the period of rapid muscle growth, resulting in the observed increased rate and efficiency of muscle deposition in implanted steers.     

Effects of pituitary adenylate cyclase-activating polypeptide (PACAP), prostaglandin E2 (PGE2) and growth hormone releasing factor (GRF) on the release of growth hormone from cultured bovine anterior pituitary cells in vitro

The effect of pituitary adenylate cyclase-activating polypeptide (PACAP) on growth hormone (GH) release was compared with that of prostaglandin E₂ (PGE₂) and growth hormone releasing factor (GRF) from cultured bovine anterior pituitary cells in vitro. Both PACAP and PGE₂ stimulated GH release at concentrations as low as 10⁻⁹ and 10⁻⁸ M, respectively, (P<0.01). However, GRF released GH at a concentration as low as 10⁻¹³ M (P<0.01). Percent increases of GH compared with controls were not significantly different among GRF, PACAP, and PGE₂ at 10⁻⁷ M; however, the increases of GH by the 10⁻⁸ M GRF, PACAP and PGE₂ were 196, 118, and 27%, respectively, (P<0.01), and 124, 65, and 1% in the 10⁻⁹ M media, respectively, (P<0.01). When GRF and somatostatin (SS) were added together, the GH releasing effect of GRF was blunted (P<0.01). Similar bluntness were observed in PACAP and PGE₂, when SS was added. The stimulatory effects of GRF and PGE₂ together were similar to that by either GRF or PGE₂ alone. When GRF and PACAP were added together, the GH released by both secretagogues was greater than that by PACAP alone (P<0.01); however, a synergistic effect was not clear when compared with GRF alone.

These findings suggest that PACAP and PGE₂ may modulate the release of GH in cattle.     

Growth hormone stimulates protein synthesis in bovine skeletal muscle cells without altering insulin-like growth factor-I mRNA expression

Growth hormone is a major stimulator of skeletal muscle growth in animals, including cattle. In this study, we determined whether GH stimulates skeletal muscle growth in cattle by direct stimulation of proliferation or fusion of myoblasts, by direct stimulation of protein synthesis, or by direct inhibition of protein degradation in myotubes. We also determined whether these direct effects of GH are mediated by IGF-I produced by myoblasts or myotubes. Satellite cells were isolated from cattle skeletal muscle and were allowed to proliferate as myoblasts or induced to fuse into myotubes in culture. Growth hormone at 10 and 100 ng/mL increased protein synthesis in myotubes (P < 0.05), but had no effect on protein degradation in myotubes or proliferation of myoblasts (P > 0.05). Insulin-like growth factor-I at 50 and 500 ng/mL stimulated protein synthesis (P < 0.01), and this effect of IGF-I was much greater than that of GH (P < 0.05). Besides stimulating protein synthesis, IGF-I at 50 and 500 ng/mL also inhibited protein degradation in myotubes (P < 0.01), and IGF-I at 500 ng/mL stimulated proliferation of myoblasts (P < 0.05). Neither GH nor IGF-I had effects on fusion of myoblasts into myotubes (P > 0.1). These data indicate that GH and IGF-I have largely different direct effects on bovine muscle cells. Growth hormone at 10 and 100 ng/mL had no effect on IGF-I mRNA expression in either myoblasts or myotubes (P > 0.1). This lack of effect was not because the cultured myoblasts or myotubes were not responsive to GH; GH receptor mRNA was detectable in them and the expression of the cytokine-inducible SH2-containing protein (CISH) gene, a well-established GH target gene, was increased by GH in bovine myoblasts (P < 0.05). Overall, the data suggest that GH stimulates skeletal muscle growth in cattle in part through stimulation of protein synthesis in the muscle and that this stimulation is not mediated through increased IGF-I mRNA expression in the muscle.     

Postnatal changes in circulating concentrations of growth hormone, somatomedin C and thyroid hormones in pigs

Plasma concentrations of growth hormone (GH), somatomedin (SmC), thyroxine (T₄) and triiodothyronine (T₃) were determined from birth to post weaning in pigs (Yorkshires). Plasma samples were obtained from the smallest, median and largest piglet from 11 litters. No differences in the circulating concentrations of any of the hormones were observed between piglets of different sizes. However, there were changes in circulating concentrations of hormones during postnatal development. Plasma concentrations of GH decreased between 2 and 8 to 10 days of age. A progressive increase in the circulating concentrations of SmC was observed with concentrations rising (3.83-fold between 2 days and 40 days of age). Plasma concentrations of T₄ and T₃ were maximal at 23 and 16 days of age, respectively.     

Dose effect of human growth hormone-releasing factor and thyrotropin-releasing factor on hormone concentrations in lactating dairy cows

Two experiments were conducted to study the effects of growth hormone-releasing factor (GRF) and thyrotropin-releasing factor (TRF) administration on hormone concentrations in dairy cows. In the first trial, 12 cows were used on 5 consecutive days to determine the effect of four sc doses of GRF (0, 1.1, 3.3 and 10 μg•kg⁻¹ BW) and three sc doses of TRF (0, 1.1 and 3.3 μg•kg⁻¹ BW) combined in a factorial arrangement. GRF and TRF acted in synergy (P = .02) on serum growth hormone (GH) concentration even at the lowest dose tested and GH response to the two releasing factors was higher than the maximal response observed with each factor alone. TRF increased (P<.01) prolactin (Prl), thyrotropin (TSH), triiodothyronine (T₃) and thyroxine (T₄) concentrations similarly at the 1.1 and 3.3 μg•kg⁻¹ doses and GRF did not interact (P>.40) with TRF on the release of these hormones. In the second trial, the effect of GRF (3.3 μg•kg⁻¹ BW, sc) and TRF (1.1 μg•kg⁻¹ BW, sc) was tested at three stages (18, 72 and 210 days) of lactation on serum Prl and TSH concentrations. Eighteen cows (n = 6 per stage of lactation) were used in two replicates of a 3 × 3 latin square. The TRF and GRF-TRF treatments were equipotent (P>.05) in increasing Prl and TSH concentrations. Prl and TSH responses were similar (P>.40) throughout lactation. In summary, GRF at doses ranging from 1.1 to 10.0 μg•kg⁻¹ and TRF at doses ranging from 1.1 to 3.3 μg•kg⁻¹ act in synergy on GH release and do not interact on Prl, TSH, T₃ and T₄ concentrations in dairy cows. Furthermore, Prl and TSH response to TRF are not affected by stage of lactation.     

Effects of VIP and GRF on the release of growth hormone in perifused bovine adenohypophysis

The effects of vasoactive intestinal polypeptide (VIP) and growth hormone releasing factor (GRF:hpGRF(1–29)-NH₂) on the release of growth hormone (GH) from anterior pituitaries from cows were examined by using an in vitro superfusion system. The pituitaries were excised randomly from cycling cows, dissected to obtain medial portions, and minced to obtain cubes with approximate dimensions of 1.5mm on a side. For each perifusion setup, 5 pieces of pituitary tissues were chambered and flushed with modified KRB solution saturated with 95% O₂-5% CO₂ at 38C. Perifusion with media containing 10⁻⁶ and 10⁻⁷M VIP for 30 min induced a significant release of GH during the treatments (P<0.05). VIP (10⁻⁸M) increased GH levels significantly (P<0.05), but to a minor degree. Perifusion with the media containing 10⁻⁶, 10⁻⁷ and 10⁻⁸M GRF for 30 min markedly increased the GH concentration and the effects continued up to 90 min after termination of the perifusion of the peptide (P<0.05, P<0.01). The GH releasing effects of GRF could be seen at doses as low as 10⁻¹¹M GRF (P<0.05, P<0.01).

These findings indicate that the GH releasing effect of VIP is less potent that that of GRF in cows.     

Hormonal changes following an acute stress in control and somatostatin-immunized pigs

Sixteen Yorkshire pigs (49 ± 2 kg BW at 17 weeks) were immunized against somatostatin (SRIF; 4 males, 4 females) or its conjugated protein, bovine serum albumin (BSA; controls; 4 males, 4 females). Immunizations were done at 10, 12 and 14 weeks of age. Jugular vein cannulae were surgically inserted at 17 weeks of age. Five d later, half of each sex from the control and SRIF-immunized groups were stressed. The other half were subjected to the same stress 48 hr later. On both days, remaining animals were used as unstressed controls. The stress consisted of 5 min of snare restraint. Blood samples were collected from all pigs on both days at −20, −15, −10, −5, 0 (beginning of stress), 2, 6, 10, 15, 20, 30, 40, 60, 90, 120, 150, 180 and 240 min. Samples were radioimmunoassayed for cortisol, growth hormone (GH), prolactin (Prl), insulin, triiodothyronine (T₃), thyroxine (T₄) and insulin-like growth factor I (IGF-I). Mean antibody titers against SRIF (1:150 dilution) at 15 weeks were 0.49 ± .09% and 54.5 ± 4.9% for control and SRIF immunized pigs, respectively. Gender and immunization against SRIF had no effect on any of the variables measured (P>0.05), except for T₃ levels which were greater in females than in males (P<0.05). The stress by time of sampling interaction was significant (P<0.01) for all hormones measured. Cortisol values almost tripled within 15 min of stress, reaching concentrations above 100 ng/mL. Maximal increases were seen at 2 min for T₄ (14%), at 6 min for T₃ (36%), at 15 min for Prl (46%) and at 10 min for insulin (141%). An increase of 129% in GH concentration was present at 20 min in stressed pigs; however, an increase of 97% was also seen at 120 min in control pigs. Concentrations of IGF-I decreased (21%) by 60 min in the stressed pigs and remained depressed for up to 150 min. Stress associated with snare restraint, therefore, induces major changes in the concentrations of a series of hormones in growing pigs. On the other hand, immunization against SRIF did not alter any of the hormonal profiles measured. Since snare restraint is widely used to handle pigs during jugular puncture, any study of hormonal secretion in this species should be carried out under carefully controlled conditions in terms of blood sampling technique.