James L. Sartin

Effect of endotoxin on pituitary hormone secretion in sheep.

Endotoxin, a potent stimulator of the immune system and an important mediator in the pathophysiology of septic shock, has been shown to alter the release of certain hormones following its systemic administration. The purpose of this study was to determine the effects of endotoxin on pituitary hormone secretion both in vivo and in vitro in sheep, with emphasis placed on its effects on growth hormone (GH) release. Endotoxin (400 ng/kg i.v.) increased plasma GH, adrenocorticotropic hormone (ACTH), cortisol and prolactin, while it decreased luteinizing hormone (LH) pulse frequency (p < 0.05). Plasma levels of tumor necrosis factor, a major mediator of endotoxin effects, also increased following endotoxin administration. Endotoxin did not affect the GH response to human GH-releasing hormone. In vitro studies evaluated the effect of endotoxin to alter GH secretion from dispersed sheep anterior pituitary cells at dosages of 1, 10 and 50 micrograms/ml, with samples collected at 4, 8 and 24 h. Endotoxin increased pituitary GH secretion at 24 h for 1 microgram/ml (p < 0.05) and at all time periods for 10 and 50 micrograms/ml (p < 0.05). It also led to an increased release of ACTH and LH in vitro. The results of this study demonstrate the ability of endotoxin to alter pituitary hormone secretion both in vivo and in vitro in sheep, suggesting a direct effect of endotoxin on the pituitary gland.

Modulation of Growth Hormone-Releasing Factor Stimulated Growth Hormone Secretion by Plasma Glucose and Free Fatty Acid Concentrations in Sheep

Effects of plasma glucose and free fatty acid (FFA) concentrations on bovine growth hormone-releasing factor (bGRF)-induced release of growth hormone (GH) were examined in ovariohysterectomized sheep. In experiment 1, the effects of an infusion of insulin (0.025 U/kg BW.h-1), glucose (40 mg/kg BW.h-1), insulin plus glucose or saline on the subsequent effects of bGRF on plasma GH concentrations were determined. Insulin-induced hypoglycemia inhibited GRF effects on plasma GH concentrations while glucose infusion enhanced bGRF actions. Infusing a higher glucose dose (120 mg/kg BW.h-1) had no effect on GRF actions. Subsequently, infusion of FFA (0.25 g/kg/.h-1), nicotinic acid (50 mg/kg BW) or saline for 1 h prior to bGRF injection demonstrated that FFA inhibited GRF actions but FFA depletion by nicotinic acid infusion had no effect on GRF actions. Nicotinic acid (40 mg/kg BW.h-1) infused for 2 h prior to bGRF injection significantly enhanced bGRF-stimulated GH secretion. Finally, to determine whether central nervous system glucopenia produced similar effects to insulin-induced hypoglycemia, 2-deoxyglucose (500 mg) was injected into the lateral ventricle followed in 1 by the i.v. injection of bGRF. The central glucopenia produced by 2-DG inhibited GRF-stimulated GH release. These data demonstrate that decreased peripheral or central nervous system glucose availability and exogenous administration of FFA antagonized GRF-induced release of GH. And, pharmacologic depletion of circulating FFA for at least 2 h facilitated GRF-induced release of GH.

Neurotransmitter Receptor Agonists Regulate Growth Hormone Gene Expression in Cultured Ovine Pituitary Cells

Abstract The regulation of growth hormone (GH) secretion and GH mRNA content by the dopaminergic agonist, bromocriptine (BRO); the β-adrenergic agonist; isoproterenol (ISO); the α1-adrenergic agonist, methoxamine (MET); the α2-adrenergic agonist, clonidine (CLON); the serotonergic agonist, quipazine (QUIP); somatostatin (SS) and GH-releasing hormone (GHRH) were studied using cultured ovine anterior pituitary cells. Clonidine and BRO (10-6 M) inhibited basal and GHRH (10-10 M)-stimulated GH release. Bromocriptine enhanced GH mRNA content and potentiated the GHRH (10-8 M)-stimulated content of GH mRNA, while CLON had no effect on GH mRNA. Quipazine had little effect on GH secretion and no effect on GH mRNA content. Methoxamine and ISO (10-6 M) increased basal secretion of GH and both enhanced GHRH-stimulated GH secretion. Both MET and ISO increased GH mRNA content of cultured ovine pituitary cells. Somatostatin (10-7 M) inhibited GHRH-stimulated GH secretion and GH mRNA accumulation. These results support the hypothesis that neurotransmitters may regulate or interact to further modulate pituitary hormone release. Moreover, the data indicate that neurotransmitters may not only regulate secretion but also regulate GH mRNA content and thus affect hormone synthesis.

Alterations in Insulin and Glucagon Secretion by Monosodium Glutamate Lesions of the Hypothalamic Arcuate Nucleus

This study was performed to determine the effects of monosodium glutamate (MSG) induced lesions of the hypothalamic arcuate nucleus (ARC) on glucose tolerance and insulin and glucagon secretion in male golden hamsters. Eight day old hamsters were given a single s.c. injection of 5.8 mg/g BW MSG or hypertonic saline (controls). Studies were initiated when the hamsters were 3 months of age. At this age there were no body weight differences. Glucose (180 mg/100 g BW) was administered via stomach tube to 18 control and 18 MSG-treated hamsters. Animals were anesthetized with ether and a single blood sample from the portal vein was taken either before or at 30 or 60 min after glucose administration (n = 6/group). Glucose concentrations were similar in both groups at all time periods. Insulin concentrations in the MSG group were significantly (P less than 0.05) elevated in MSG-treated hamsters compared to controls at the 60 min time point. Glucose suppressed glucagon (P less than 0.05) in control but not in MSG-treated hamsters. The MSG group had significantly more glucagon (P less than 0.05) in portal vein blood at 30 min after glucose administration than did the control hamsters. Molar insulin/glucagon ratios did not differ between the 2 groups which likely accounts for the lack of differences in blood glucose levels. These results suggest a role for the ARC in regulating pancreatic function.

Characterization of Canine Triiodothyronine (T3) Autoantibodies and Their Effect on Total T3 in Canine Serum

Abstract A study of 3,5,3′-L-triiodothyronine autoantibody (T3 AA) in 18 dogs revealed an average apparent affinity constant for T3 of 2.24 ± 1.78 × 1010 M -1, an average T3 binding capacity of 639.3 ± 666.5 ng/dl and a low thyroxine (T4) cross-reactivity (<1%) in all samples tested. A valid radioimmunoassay (RIA) procedure which involved heat treatment of samples for 1 hr at 70°C and assay on Sephadex minicolumns was developed for measuring T3 in the presence of T3 AA. Total T3 was elevated (x = 374.8 ± 158.4 ng/dl) in samples in which T4 was in the normal canine range, but T3 was lower (x = 96.1 ± 63.3 ng/dl) in samples with T4 values in the hypothyroid range. For each sample the concentration of T3 not bound by T3 AA was calculated from the total T3 concentration, the affinity constant, and the binding capacity. In dogs with normal total T4 concentrations the average calculated T3 not bound by T3 AA was 147.2 ± 144.4 ng/dl while in dogs with low total T4 the value was 15.7 ± 26.3 ng/dl (normal canine range is 45-150 ng/dl). Canine samples containing T3 AA were compared to serum from three rabbits actively immunized against T3 to provide anti-T3 for commercial RIA. The rabbit T3-antisera had an average T3 affinity constant similar to those of the canine samples (1.57 × 1010 M -1), but had average titer, T3 binding capacity, and total T3 values more than 10-fold higher. Our findings indicate that, in dogs with serum containing T3 AA and normal total T4 concentrations, a compensatory mechanism appears to exist to maintain non-T3 AA bound T3 within the range of normal total T3. This compensatory mechanism does not operate in those dogs with insufficient thyroid activity to maintain normal total T4 values.

Evaluation of endocrine and immune responses of steers challenged with infectious bovine rhinotracheitis virus

OBJECTIVEnTo evaluate the endocrine and immune responses of steers challenged with infectious bovine rhinotracheitis virus (IBRV).nnnANIMALSn12 crossbred beef steers.nnnPROCEDURESnSteers were randomly assigned to IBRV- (control) or IBRV+ treatment groups. Experimentally challenged steers (IBRV+) received a dose of IBRV intranasally (8.0 50% tissue culture infective doses), IBRV- steers received a saline (0.9% NaCl) solution placebo intranasally, and each group was placed in an isolated paddock. At 72 hours after challenge, all steers were fitted with indwelling jugular catheters and placed into individual stanchions. Blood samples were collected on days 4 through 8. Serum was analyzed for concentrations of cortisol, interleukin-6, interferon-γ, tumor necrosis factor-α, growth hormone, and insulin-like growth factor I.nnnRESULTSnFrom 72 to 144 hours after challenge inoculation, the IBRV+ group had significantly greater mean rectal temperature, compared with the IBRV- group; the greatest temperatures in both groups were observed at 72 hours. Serum cortisol concentrations were increased in both groups from hours 72 to 136 and serum interferon-γ concentrations were greater in the IBRV+ from 94 to 112 hours after inoculation. Growth hormone concentration was greater in the IBRV+ group at various time points, but no difference in insulin-like growth factor- I concentration was observed.nnnCONCLUSIONS AND CLINICAL RELEVANCEnResults indicated that IBVR challenge altered growth hormone concentration at some time points but was not associated with large increases in circulating proinflammatory cytokines.