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The effects of exercise on the molecular nature of secreted human growth hormone (GH) or its biological activity are not well understood. Plasma from women (average age 23.6 yr, n = 35), drawn before and after an acute heavy resistance exercise test, was fractionated by size exclusion chromatography into three size classes, namely, > 60 kDa (fraction A), 30-60 kDa (fraction B), and < 30 kDa (fraction C), before GH assay. Concentrations of GH in these fractions, as well as in unfractioned plasma, were measured by the Nichols immunoradiometric assay, National Institute of Diabetes and Digestive and Kidney Diseases (NIDDK) polyclonal competitive RIA, Diagnostic Systems Laboratory's immunofunctional assay (measures dimerization-capable species), and the rat tibial bioassay. Significantly increased circulating GH concentrations of two- to fourfold were observed when immunoassays in unfractionated plasma samples were used, but they showed no significant change with use of the rat tibial bioassay. Significant exercise-induced increases in GH were found in fractions B and C but not in fraction A. Because chemical reduction of the samples before GH immunoassay significantly increased GH concentrations in fractions B and C (Nichols and NIDDK kits) after exercise, it is concluded that exercise may specifically increase release of disulfide-linked hormone molecules and/or fragments. Finally, because most of the GH released after exercise was able to dimerize the GH receptor in vitro, it is also concluded that these forms have the two intact binding sites required to initiate signal transduction in target cells.

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This study examined the hypothesis that exercise-induced changes in circulating testosterone would be centrally mediated via hypothalamic-pituitary release of luteinizing hormone (LH). We tested this hypothesis by examining overnight LH, total and free testosterone (TT and FT), and cortisol (C) concentrations in 10 young healthy men (21 +/- 1 yr) during two experimental sessions: a control and an acute heavy-resistance exercise bout (50 total sets consisting of squats, bench press, leg press, and latissimus dorsi pull-down). Exercise was performed from 1500 to 1700, and blood sampling began at 1700 and continued until 0600 the next morning. Blood was sampled every 10 min for LH and every hour for TT, FT, and C. Hormonal concentrations were determined via RIA, and the secretion characteristics of LH were analyzed with deconvolution analysis. When overnight postexercise concentrations were compared with control concentrations, no statistically significant (P < or = 0.05) differences were observed for LH half-life, LH pulse frequency, interpulse interval, pulse amplitude, or pulse mass. Significant differences were observed for LH production rate (13.6 +/- 4 and 17.9 +/- 5 IU. l distribution volume(-1) x day(-1) for exercise and control, respectively, a 24% reduction). For the ANOVA marginal main effect means due to condition, C was significantly elevated (5.9 +/- 0.7 vs. 4.0 +/- 0.4 microg/dl), while TT (464 +/- 23 vs. 529 +/- 32 ng/dl) and FT (15.6 +/- 0.7 vs. 18.3 +/- 0.9 pg/ml) were significantly decreased for the exercise condition. These data demonstrate that the decline in overnight testosterone concentrations after acute heavy-resistance exercise is accompanied by a blunted LH production rate and elevated C concentrations.

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This investigation examined the hypothesis that acute heavy resistance exercise (AHRE) would increase overnight concentrations of circulating human growth hormone (hGH). Ten men (22 +/- 1 yr, 177 +/- 2 cm, 79 +/- 3 kg, 11 +/- 1% body fat) underwent two overnight blood draws sampled every 10 min from 1700 to 0600: a control and an AHRE condition. The AHRE was conducted from 1500 to 1700 and was a high-volume, multiset exercise bout. Three different immunoassays measured hGH concentrations: the Nichols immunoradiometric assay (Nichols IRMA), National Institute of Diabetes and Digestive and Kidney Diseases radioimmunoassay (NIDDK RIA), and the Diagnostic Systems Laboratory immunofunctional assay (DSL IFA). The Pulsar peak detection system was used to evaluate the pulsatility profile characteristics of hGH. Maximum hGH was lower in the exercise (10.7 microg/l) vs. the control (15.4 microg/l) condition. Mean pulse amplitude was lower in the exercise vs. control condition when measured by the Nichols IRMA and the DSL IFA. A differential pattern of release was also observed after exercise in which hGH was lower in the first half of sleep but higher in the second half. We conclude that AHRE does influence the temporal pattern of overnight hGH pulsatility. Additionally, because of the unique molecular basis of the DSL IFA, this influence does have biological relevance because functionally intact molecules are affected.

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The aim of this study was to determine the impact of resistance exercise on neuromuscular junction (NMJ) architecture. Eighteen Sprague-Dawley rats either participated in a 7-week resistance training program or served as untrained controls. Following the experimental period, the NMJs of soleus muscles were visualized with immunofluorescent techniques, and muscle fibers were stained histochemically. Results indicate that resistance training significantly (P < 0.05) increased endplate perimeter length (15%) and area (16%), and significantly enhanced the dispersion of acetylcholine receptors within the endplate region. Pre- and post-synaptic modifications to resistance exercise were well-coupled. No significant alterations in muscle fiber size or fiber type were detected. The data presented here indicate that the stimulus of resistance training was sufficiently potent to remodel NMJ structure, and that this effect cannot be attributed to muscle fiber hypertrophy or fiber type conversion.

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Somatotropin (ST) administration enhances protein deposition and elicits profound metabolic responses, including hyperinsulinemia. To determine whether the anabolic effect of ST is due to hyperinsulinemia, pair-fed weight-matched growing swine were treated with porcine ST (150 microg x kg body wt(-1) x day(-1)) or diluent for 7 days (n = 6/group, approximately 20 kg). Then pancreatic glucose-amino acid clamps were performed after an overnight fast. The objective was to reproduce the insulin levels of 1) fasted control and ST pigs (basal insulin, 5 microU/ml), 2) fed control pigs (low insulin, 20 microU/ml), and 3) fed ST pigs (high insulin, 50 microU/ml). Amino acid and glucose disposal rates were determined from the infusion rates necessary to maintain preclamp blood levels of these substrates. Whole body nonoxidative leucine disposal (NOLD), leucine appearance (R(a)), and leucine oxidation were determined with primed, continuous infusions of [(13)C]leucine and [(14)C]bicarbonate. ST treatment was associated with higher NOLD and protein balance and lower leucine oxidation and amino acid and glucose disposals. Insulin lowered R(a) and increased leucine oxidation, protein balance, and amino acid and glucose disposals. These effects of insulin were suppressed by ST treatment; however, the protein balance remained higher in ST pigs. The results show that ST treatment inhibits insulin's effects on protein metabolism and indicate that the stimulation of protein deposition by ST treatment is not mediated by insulin. Comparison of the protein metabolic responses to ST treatment during the basal fasting period with those in the fully fed state from a previous study suggests that the mechanism by which ST treatment enhances protein deposition is influenced by feeding status.

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Somatotropin (ST) administration enhances protein deposition in well-nourished, growing animals. To determine whether the anabolic effect is due to an increase in protein synthesis or a decrease in proteolysis, pair-fed, weight-matched ( approximately 20 kg) growing swine were treated with porcine ST (150 microg. kg(-1). day(-1), n = 6) or diluent (n = 6) for 7 days. Whole body leucine appearance (R(a)), nonoxidative leucine disposal (NOLD), urea production, and leucine oxidation, as well as tissue protein synthesis (K(s)), were determined in the fed steady state using primed continuous infusions of [(13)C]leucine, [(13)C]bicarbonate, and [(15)N(2)]urea. ST treatment increased the efficiency with which the diet was used for growth. ST treatment also increased plasma insulin-like growth factor I (+100%) and insulin (+125%) concentrations and decreased plasma urea nitrogen concentrations (-53%). ST-treated pigs had lower leucine R(a) (-33%), leucine oxidation (-63%), and urea production (-70%). However, ST treatment altered neither NOLD nor K(s) in the longissimus dorsi, semitendinosus, or gastrocnemius muscles, liver, or jejunum. The results suggest that in the fed state, ST treatment of growing swine increases protein deposition primarily through a suppression of protein degradation and amino acid catabolism rather than a stimulation of protein synthesis.

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Ectopic expression of a new serum protease-resistant porcine growth hormone-releasing hormone, directed by an injectable muscle-specific synthetic promoter plasmid vector (pSP-HV-GHRH), elicits growth in pigs. A single 10 mg intramuscular injection of pSP-HV-GHRH DNA followed by electroporation in three-week-old piglets elevated serum GHRH levels by twofold to fourfold, enhanced growth hormone secretion, and increased serum insulin-like growth factor-I by threefold to sixfold over control pigs. After 65 days the average body weight of the pigs injected with pSP-HV-GHRH was approximately 37% greater than the placebo-injected controls and resulted in a significant reduction in serum urea concentration, indicating a decrease in amino acid catabolism. Evaluation of body composition indicated a uniform increase in mass, with no organomegaly or associated pathology.

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During the past 15 years, many investigators have examined the effects of somatotropin (ST) on growth and lactation in farm animals. Throughout this period, concerns about potential effects of ST on reproduction have been expressed. The objective of the present review will be to focus on the effects of exogenous ST on the hypothalamic-pituitary-gonadal axis. Plasma progesterone is increased when recombinant bovine (rb)ST is given to cattle, early in lactation, and also to sheep. Also, the release of progesterone from cultured swine and human luteal cells is increased by ST. Treatment with rbSt increases the numbers of small follicles, but does not increase the ovulatory rate of ruminants. Doses of ST similar to those used to increase milk production do not affect the secretion of testosterone or spermatogenesis in rams or bulls. Stimulatory and inhibitory effects of exogenous ST on reproductive function of gilts have been reported. Daily injections of porcine ST (pST) delayed puberty and expression of estrus after puberty. Daily administration of pST increased the number of small follicles, but not of medium follicles, whereas administration of pST by using a sustained release implant increased the number of medium follicles. Size and weight of reproductive organs and concentration of testosterone are not affected when pST is administered for at least 42 d. However, pST enhanced testicular development and spermatogenesis when given to neonatal boars. In summary, administration of exogenous ST at doses known to alter milk production and carcass composition may have subtle positive and/or negative effects on the reproductive systems of cattle and swine; however, these effects appear to be transient.

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The purpose of these studies was to determine the effect of thyroidectomy (Tx), and thyroid hormone (T3/T4) treatment on concentrations of plasma CT in chicks. In addition, the turnover of CT in Tx- and T3/T4-treated chicks was estimated using a novel nonradioactive salmon CT preparation. One-week-old broiler chicks (Gallus domesticus) (n = 75) were divided into three groups. Group I was sham-injected daily (i.m. saline), Group II was injected with 50 micrograms/day of T3/T4 while Group III was injected with the goitrogen, methimazole, (150 mg/kg BW per day) for 8 weeks. Chicks (8-9 weeks old) were implanted with catheters in the brachial wing vein and administered ruthenium-labeled salmon CT. Blood samples were collected at 30 s, 1, 2, 4, 8, 20 min, and 3 h after injection. Results showed that concentrations of plasma CT were decreased in T3/T4-injected birds. There was no significant effect of methimazole on circulating concentrations of plasma CT. The half-life of CT was significantly increased (P < 0.05) in both T3/T4-injected (n = 6; 1.34 +/- 0.16 min) and goitrogen-treated birds (n = 2; 5.81 +/- 2.83 min) compared to controls (n = 7; 54 +/- 3 s) The results demonstrate that changes in concentrations of plasma thyroid hormones can significantly affect concentrations of plasma CT.

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The objective of the present study is to apply a novel side-view imaging technique to investigate T-leukemic Jurkat cell adhesion to a surface-immobilized ICAM-1 in shear flow, a ligand for leukocyte LFA-1. Images have revealed that Jurkat cell adhesion on ICAM-1 under flow conditions in vitro is quasistatic. The cell-substrate contact length steadily increased with time during the initial cell attachment to the ICAM-1-coated surface and subsequently decreased with time as the trailing edge of the cell membrane peeled away from the substrate under the influence of fluid shear forces. Changes in flow shear stresses, cell deformability, or substrate ligand strength resulted in a significant change in the characteristic adhesion binding time and contact length. A 3-D flow field with shear stresses acting on an adherent cell was calculated by using finite element methods based on cell shapes obtained from the in vitro images. The maximum shear stress acting on an actual cell body was found to be 3-5 times greater than the upstream inlet wall shear stress and was influenced by the extent of cell deformation within the flow channel. Therefore, the application of such a side-view imaging technique has provided a practical assay to study the mechanics of cell-surface adhesion in 3-D. The elongation of cells in shear flow tempers hydrodynamic shear forces on the cell, which affects the transients in cell-surface adhesion.

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BACKGROUND: A previous study revealed that exposure of rats to microgravity for 14 d on Cosmos 2044 reduced production of testosterone by > 80%, although spermatogenesis remained essentially normal. METHODS: To ascertain if testicular function was altered in hypergravity, 60-d-old rats were randomly assigned to 2 groups (10 per group) and subjected to 14 d of centrifugation to expose them to a total of 2G, or held at unit gravity in similar cages without centrifugation (control). RESULTS: After 14 d, body weight of 2G rats was essentially unchanged, whereas that of control rats had increased; 310 vs. 377 g (p < 0.05). Testes weight, production and secretion of testosterone, diameters of seminiferous tubules and their lumina, data from subjective evaluation of spermatogenesis, and counts of homogenization-resistant spermatids all were similar for 2G and control rats. CONCLUSION: It was concluded that exposure of male rats to 2G for 14 d had no major effect on testicular function whereas, based on earlier studies, exposure to microgravity (< 10(-3) x gravity) for 11-14 d suppressed production of testosterone by Leydig cells and reduced concentrations of anabolic steroids available to peripheral tissues.

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The roles of photoperiod, energy balance, and concentrations of plasma total calcium (CaT) on daily changes in plasma calcitonin (CT) were investigated in broiler breeder hens (84-100 weeks old). In the first study, broiler breeder hens (n = 24), reared on 14L:10D, were divided into two groups. One group was transferred from a restricted diet (DR) of 150 g/day to ad libitum (AL) for 14 days, while the other group remained on DR. After 2 weeks of ad libitum feeding, birds from each group (AL and DR) were bled every 2 hr for 24 hr for measurement of plasma CaT and CT. In a second study, the hens (n = 20) were transferred to continuous light (LL) for 30 days. After the 30 days, food was removed from one group for 48 hr prior to blood sampling for 24 hr at 3-hr intervals. In a third study, birds were transferred to an ahemeral light cycle (11L:10D) for 28 days. Food was removed from the birds (n = 11) for 48 hr prior to blood sampling every 3 hr for 24 hr. Four weeks later the same birds were bled again for 24 hr, but this time the birds were maintained on a restricted feeding schedule. Plasma CT was measured by a specific heterologous electrochemiluminescent (ECL) assay while plasma CaT was measured by atomic absorption. The results showed that plasma CT concentrations did not correlate with plasma CaT concentrations. Comparisons made between initial and final CaT and CT concentrations indicated an effect of stress due to repeated handling of the birds. Concentrations of plasma CT were significantly reduced in the fasted animals (P < 0.05) compared to diet-restricted controls. There was a significant increase in plasma CT during the dark period of fed animals which was abolished in animals maintained on LL or fasted. In conclusion, a surge in plasma CT requires that the hens be provided food and that they be exposed to a dark cycle.

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The neuroendocrine mechanisms underlying the development of pulsatile release of LH in bull calves are poorly understood. The hypothesis that endogenous excitatory amino acids, working through N-methyl-D-aspartate (NMDA) receptors, are involved in the generation of pulsatile LH release during sexual maturation of bull calves was tested. Holstein bull calves were administered i.v. 0.001, 0.01 and 0.1 mg kg-1 body mass of MK-801, a specific noncompetitive NMDA receptor antagonist, on alternate days at 1, 12 and 24 weeks of age (n = 6 per dose), using a replicated 3 x 3 latin square design. Blood samples were obtained from jugular catheters at intervals of 10 min for 2 h before and 2 h after MK-801 treatment at 1 week and for periods of 4 h before and after MK-801 administration at 12 and 24 weeks of age. Plasma concentrations of LH were measured by a specific radioimmunoassay and pulsatile LH secretion was analysed using the Pulsar algorithm. Basal LH secretion at 1 week of age was low but was increased by 12 weeks (0.38 +/- 0.01 versus 1.38 +/- 0.11 ng ml-1; P < 0.001) with the establishment of frequent high amplitude pulses. Mean LH concentrations and pulse amplitude, but not frequency, were lower at 24 than at 12 weeks of age (0.88 +/- 0.08 versus 1.38 +/- 0.11 ng ml-1: P < 0.001, 1.74 +/- 0.14 versus 2.82 +/- 0.36 ng ml-1: P < 0.01, and 1.70 +/- 0.30 versus 1.70 +/- 0.41 in 4 h: P > 0.05, respectively). Administration of MK-801 did not affect LH secretion in 1- and 12-week-old calves.(ABSTRACT TRUNCATED AT 250 WORDS)

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Chronic exercise training may alter the hypothalamic-pituitary-ovarian axis by affecting gonadotropin secretion and structural indexes of the anterior pituitary gonadotropes. Young female Harlan Sprague-Dawley rats were randomly assigned to either a 12-wk intensive treadmill-running program (n = 16) or to a sedentary untrained group [untrained-proestrus (UTP), n = 16]. After training for 3 mo, animals were killed on proestrous morning, and individual pituitary glands were enzymatically dispersed into single-cell suspensions. Trained rats maintained either predominantly regular estrous cycles [trained-proestrus (TP), n = 11] or exhibited anestrous vaginal cytology [trained-anestrus (TA), n = 5]. At death, plasma estradiol concentrations for TP (57.4 +/- 6 pg/ml, mean +/- SE) and UTP (54.6 +/- 5 pg/ml) were similar. However, TA had plasma estradiol levels (2.7 +/- 0.2 pg/ml; P < 0.05) reflecting their anestrous status. Basal luteinizing hormone (LH) secretion (20-h static incubation) was significantly reduced in gonadotropes from TA (7.8 +/- 1.1 ng/pooled wells) compared with TP (13.6 +/- 1.6 ng/pooled wells) and UTP (12.6 +/- 1.0 ng/pooled wells). Immunofluorescence LH staining and cytoplasmic granularity were significantly lower within the TA gonadotrope population compared with those of TP and UTP rats. Evidence of impaired basal LH secretion, reduced gonadotrope LH-specific fluorescein content, and reduced gonadotrope cytoplasmic granularity in TA rats suggests that, concomitant with proposed changes in the central neural drive, there are important exercise training-induced alterations of the anterior pituitary gonadotrope that influence rats' estrous cycles and affect their reproductive status.

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The objectives of this study were (i) to determine whether age-related changes in the secretion of LH are associated with alterations in secretory activity or numbers of gonadotrophs, and (ii) to determine whether gonadotrophs obtained from the pars distalis and pars tuberalis undergo similar age-related changes in function. Blood samples were collected from Holstein bull calves every 15 min for 12 h at < 1, 12, and 42 weeks of age (n = 5 per age group) to characterize the secretion of LH. Calves were killed 3-5 days later. The pars distalis and pars tuberalis were enzymatically dispersed into suspensions of single cells. Cells from the pars distalis were (i) extracted with 0.01 mol NaHCO3 l-1, (ii) fixed for immunocytochemical analysis, and (iii) cultured in six-well plastic plates at a density of 500,000 cells per well in media containing 2.5% homologous calf serum for 18 and 72 h. Cells from the pars tuberalis were cultured as for pars distalis cells. As expected, LH pulse frequency increased (P < 0.01) between one and 12 weeks of age and then declined. The percentage of cells from the pars distalis containing immunoreactive LH averaged 8.4%, and did not change with age. The mass of the pars distalis and the total number of cells recovered increased with age (P < 0.05); consequently, the number of gonadotrophs recovered also increased. The initial content of LH of pars distalis cells changed with age and was greatest at 12 weeks.(ABSTRACT TRUNCATED AT 250 WORDS)

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The influence of triiodothyronine (T3) on growth hormone (GH) mRNA and GH secretion has been examined in the chicken. Initially T3 treatment in the diet for three days did not alter plasma concentrations of GH. Plasma concentrations of GH were depressed with seven and 14 days of T3 treatment (1 or 5 ppm in the diet). There was a concomitant decline in pituitary GH mRNA with T3 treatment. Pituitary GH content was reduced with 14 but not seven days of T3 treatment. No effect of T3 was observed on the percentage by somatotroph cells in the anterior pituitary gland by fluorescence flow cytometry analysis. While exposure of pituitary cells in vitro from young chickens to GHRF for 2 hr increased GH mRNA, no effect was observed with T3. The presence of T3, for 48 hr in vitro, tended to reduce GH mRNA in adenohypophyseal cells from young chickens and decreased GH mRNA with anterior pituitary cells from adult chicks. It is concluded that T3 chronic administration of T3 depresses circulating concentrations of GH, at least in part, by decreasing GH mRNA and hence GH synthesis.

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In a 2 x 2 factorial experiment, 110 Holstein cows (55 first-breeding and 55 repeat-breeding), free of genital abnormalities, were injected i.m. at the time of insemination with either saline solution or 100 micrograms of GnRH. Blood samples were drawn from the tail vein or artery of 22 cows (10 first- and 12 repeat-breeding) immediately before GnRH injection and again 1 and 2 h later to determine whether the GnRH induced the release of LH. The GnRH caused LH release in 18 of 22 cows, with a greater (P < .05) mean concentration of LH than that in saline-treated cows at 1 h (2.3 vs 7.0 ng/mL) and 2 h (2.5 vs 6.0 ng/mL) after injection. Length of estrus and time of ovulation were calculated from estrus checks and ovarian palpations per rectum at 8-h intervals. The GnRH injections produced no change in duration of estrus (19.2 h) or time of ovulation postestrus (9.5 h). The cows were slaughtered 41 to 90 (mean = 60) h after ovulation to determine the ovum cleavage rate and the number of accessory sperm in the zona pellucida. The GnRH treatment increased the incidence of twin ovulations but did not increase the number of accessory sperm or improve the proportion of ova that cleaved in either first-service or repeat-breeding cows.

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Insulin-like growth factors (IGF) are important growth regulators in many species, and their effects are influenced by their association with IGF-binding proteins (IGFBP). The objectives of this study were to characterize the ontogeny of the blood plasma IGFBP in calves and to determine the effect of dietary IGF-I neonatal plasma IGFBP. Plasma from newborn and 7-d-old male calves fed milk replacer, milk replacer + recombinant human IGF-I (rhIGF-I), or colostrum for 2 d followed by milk replacer was analyzed for IGFBP by ligand blot analysis. In addition, plasma samples from 1-, 12-, 24-, and 45-wk-old male calves were analyzed for IGFBP and IGF-I. Newborn and 7-d-old calf plasma contained IGFBP with M(r) of 26, 34, and 42 to 48 kDa. These profiles were not affected by the dietary treatments; however, a slight increase in the 34-kDa IGFBP and a slight decrease in the 26- and 42- to 48-kDa IGFBP were detected from birth to 7 d of age. The 34-kDa was confirmed to be bovine IGFBP-2 by immunoblot and the 42- to 48-kDa is likely IGFBP-3. The 29-, 31-, and 42- to 48-kDa IGFBP increased between 1 and 45 wk of age. Similarly, plasma IGF-I concentrations were increased from 49.7 to 449.7 ng/mL in plasma from calves from 1 to 45 wk of age. In contrast, the 34-kDa IGFBP increased from 1 to 12 wk but then gradually decreased from 12 to 45 wk, whereas the 26-kDa IGFBP did not change.(ABSTRACT TRUNCATED AT 250 WORDS)

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The average concentration of GH in blood is high at birth and declines during the period of sexual maturation in bulls. The objectives of these studies were (1) to define age-related changes in vivo in the pulsatile secretion of GH from birth to puberty, (2) to determine whether pituitary cell content of GH and characteristics of the secretion of GH in vitro reflect age-related changes in vivo, and (3) to examine whether responsiveness to GH-releasing hormone (GHRH) and somatostatin (SRIF) in vitro changed with age in Holstein bull calves. In experiment 1, calves were bled every 15 min for 12 h at < 1, 12 and 42 weeks of age (n = 5/group), these being representative of infantile, juvenile and pubertal stages of development. Calves were killed 3 to 5 days later and the pars distalis of the anterior pituitary gland was enzymatically dispersed into a suspension of single cells. Aliquots of cells were extracted with 0.01 mol NaHCO3/l to determine the content of GH and cultured for 18 and 72 h. As expected, the average concentration of GH in plasma decreased with age (P < 0.001). The initial decrease in GH was caused by a reduction in the baseline concentration between birth and 12 weeks of age. There was a marked decrease in GH pulse amplitude between 12 and 42 weeks of age and a further reduction in the baseline concentration. In contrast, the pulse frequency of GH increased (P < 0.05) from < 1 week to 12-weeks of age and remained constant thereafter.(ABSTRACT TRUNCATED AT 250 WORDS)

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