Inhibitory effect of calcitonin on growth hormone and insulin secretion in man

The effect of calcitonin administration on basal and arginine-stimulated growth hormone and insulin plasma levels was investigated. The intramuscular injection of synthetic salmon calcitonin (100 U MRC) in five normal subjects produced a significant decrease (p less than 0.05) in insulin concentration. The same amount of calcitonin given 15 min before an arginine infusion test in seven normal subjects significantly reduced the response of growth hormone (p less than 0.025) and insulin (p less than 0.005) to the stimulus.     

Effects of neonatal administration of monosodium glutamate on somatotrophs and growth hormone secretion in prepubertal male and female rats

Female and male rats were injected with monosodium L-glutamate (MSG; 4 mg/g BW) or 0.9% saline as neonates and then decapitated on days 35 and 40 of life, respectively. Trunk blood was collected for RIA of serum GH. Anterior pituitary glands (APGs) were bisected. One half was assayed for GH. The other half was placed in culture medium to study the basal GH release rate. Pituitary sections from additional rats were stained for GH, and morphometric analyses were performed on the GH cells. Treatment with MSG lowered serum GH levels and gland GH content in female but not in male rats. MSG did not alter the gland GH concentration or the basal GH release rate whether expressed per mg APG or per entire gland in either sex. The mean cross-sectional area of GH cells was reduced in either sex of MSG-treated rats. The numerical density of GH cells and the percentage of GH cells in APGs were similar in saline- and MSG-treated rats of either sex. The volume density of GH cells was lower in MSG-treated male rats only. The results suggest that in prepubertal rats which had been given MSG as neonates there is a sex difference in mean serum GH concentration and APG GH content, GH cell size is reduced in both sexes, and the individual GH cells contain normal amounts of GH in spite of their smaller size.     

Influence of endogenous somatostatin on growth hormone and thyrotropin secretion in neonatal rats

When gestating rats were injected iv with an antiserum to somatostatin (SRIF-AS) during the last week of gestation, serum GH levels in fetuses and 6-h-old newborn rats were not significantly different from controls. Similarly, 2 h after the ip administration of SRIF-AS, no change in serum GH concentration was observed in 2-h-old rats. However, under the same conditions, a significant increase in serum GH was observed in 24-h-old rats and in 2- to 60-day-old rats. The injection of SRIF-AS neither changed basal serum TSH levels during the neonatal development nor in the adult stage. A significant increase in TRH-induced TSH release was observed after the third postnatal day. It is concluded that endogenous SRIF plays a physiological role in GH release by 24 h of age in the rat and that the fall in GH secretion that normally occurs during the first week of life is due to the development of inhibitory mechanisms mediated by hypothalamus SRIF. Additionally the results suggest that the influence of SRIF upon TSH secretion is present before that of TRH.     

Differential effects of neonatal and adult androgen exposure on the growth hormone secretory pattern in male rats

The interactive effects of androgen exposure during neonatal and adult life on the pattern of GH secretion in adult male rats was investigated. Neonatal rats were orchidectomized or sham-operated on days 1-2 of life and injected immediately postoperatively with testosterone propionate (250 micrograms, sc) or vehicle. At 90-130 days of age the rats were bled every 20 min between 9 and 17 h from an indwelling intraatrial catheter. Some neonatally gonadectomized, testosterone- or vehicle-treated rats were also given depot testosterone (15 mg/kg, im) 5-10 days before blood sampling. Plasma GH concentrations were measured by RIA, and the pulsatile secretory patterns were analyzed by the PULSAR computer program. Neonatal orchidectomy resulted in a marked suppression (50-75%) of both the height and duration of GH secretory episodes, while baseline GH levels were higher in neonatally gonadectomized males than in sham-operated controls. Neonatal testosterone replacement therapy restored high amplitude GH pulses. However, the GH pulses of these animals were of significantly shorter duration and occurred more frequently, and baseline GH levels were markedly higher than those in intact male rats. In contrast, neonatally gonadectomized rats treated with testosterone both neonatally and during adulthood exhibited a GH pattern indistinguishable from that in normal males, with high amplitude and long-lasting (103 +/- 8 min) pulses at regular intervals (178 +/- 9 min). A similar masculine GH pattern was seen in neonatally gonadectomized rats given testosterone only during adult life. The present results indicate that high amplitude GH pulses can be induced by either neonatal or adult androgen exposure. However, while neonatal androgens irreversibly cause stimulation of overall GH secretion, only the continuous presence of androgens during adult life can induce a GH secretory pattern, consisting of large surges at regular 3-h intervals separated by a low baseline that is characteristic of normal male rats.     

The effect of growth hormone replacement therapy on adrenal androgen secretion in adult onset hypopituitarism

OBJECTIVE: Growth hormone replacement therapy in GH-deficient children is associated with enhanced adrenal androgen production, raising the possibility that GH might stimulate adrenocortical hormone secretion. This has not been extensively investigated in adults to date. GH is a potent modulator of the activity of the 11beta-hydroxysteroid dehydrogenase type 1 (11beta-HSD1) enzyme and by altering cortisol metabolism can affect the function of the hypothalamo-pituitary-adrenal (HPA) axis and therefore potentially of adrenal androgen secretion. This study examined the effects of GH replacement in GH-deficient adults on adrenal androgen secretion. DESIGN: Prospective study of the effect of GH replacement therapy on adrenal androgen production in patients with adult onset hypopituitarism over a 12-month period. PATIENTS AND METHODS: Thirty adult GH-deficient patients were classified into two groups according to their cortisol responses to an insulin-induced hypoglycaemia or a glucagon stimulation test: 13 patients were adrenocorticotropic hormone (ACTH)-sufficient (nine females, age 45.1 +/- 3 years), whereas 17 patients were ACTH-deficient (11 females, age 45.5 +/- 3 years). Serum samples were collected before patients were initiated on GH replacement therapy using a dose titration regimen, and after 6 and 12 months on GH therapy for measurement of serum IGF-I, dehydroepiand-rosterone sulphate (DHEAS), Delta4-Androstenedione (A4), testosterone, cortisol, sex hormone binding globulin (SHBG) and cortisol binding globulin (CBG). RESULTS: Six months after the initiation of GH replacement therapy, serum IGF-I levels were within the normal age-related reference range in both groups of patients and this was maintained at 12 months [in all patients 0 vs. 6 months: median (interquartile range): 92.5 ng/ml (73-116 ng/ml) vs. 191 ng/ml (159-224 ng/ml), P < 0.01]. In both ACTH-sufficient and -deficient groups of GH-deficient patients, pretreatment serum DHEAS levels were lower than the normal age-related reference range (P < 0.01); the ACTH-deficient patients had significantly lower DHEAS levels than the ACTH-sufficient patients [median (interquartile range): 0.5 micro mol/l (0.4-1.2 micro mol/l) vs. 1.5 micro mol/l (0.6-2.7 micro mol/l), P < 0.05]. Following GH replacement therapy, median levels of serum DHEAS levels rose from 1.5 micro mol/l (0.6-2.7 micro mol/l) to 1.9 micro mol/l (1.9-3.9 micro mol/l) in ACTH-sufficient patients, increasing in 11 of the 13 patients (P < 0.02). In this group, the median percentage increase from baseline was 32% at 6 months (P < 0.05). In contrast, baseline serum DHEAS levels [0.5 micro mol/l (0.4-1.2 micro mol/l)] declined in or from the measurable range in 47% of ACTH-deficient patients [median -16%; range -36-0] and only in one patient a + 0.2 micro mol/l increase was observed. GH dose requirements tended to be lower in ACTH-sufficient patients [1.2 U/day (0.8-1.4 U/day) vs. 1.6 U/day (1.0-2.0 U/day); P = 0.062]. There were no significant changes in serum testosterone, A4, SHBG and/or CBG levels, compared to the pretreatment levels, in either group of patients over the 12 months of GH replacement. CONCLUSIONS: This study shows that median serum DHEAS levels are significantly lower in GH-deficient patients, even those with intact ACTH reserve, than in aged-matched controls. GH replacement therapy is associated with a significant increase in mean serum DHEAS only in ACTH-sufficient patients. These findings are consistent with either (i) GH stimulation of adrenal androgen production in the permissive presence of ACTH or (ii) an inhibitory effect of GH on 11beta-HSD type 1 activity leading to enhanced cortisol clearance, subsequent activation of the HPA axis and ACTH-mediated androgen secretion.     

Depletion of hypothalamic growth hormone-releasing hormone by neonatal monosodium glutamate treatment reveals an inhibitory effect of betamethasone on growth hormone secretion in adult rats

Rats were treated with monosodium glutamate (MSG), 4 mg/g on alternate days for the first 10 days of life, to induce lesions of the arcuate nucleus and destroy the majority of growth hormone-releasing hormone (GHRH) neurones. At 10 weeks of age, control (n = 42) and MSG-treated (n = 36) male rats were used to test the effect of glucocorticoids on growth hormone (GH) secretion. Each treatment group was divided into six study groups to determine the effect of betamethasone (BM), administered either 3 or 20 h prior to sacrifice, alone and in combination with hypoglycaemia (insulin 0.1 U/100 g). BM treatment of male rats was without effect on plasma GH levels in control animals. In contrast, glucocorticoid administered either 3 h before sacrifice or the previous evening significantly reduced circulating GH (p less than 0.001) in MSG-treated animals. The difference in plasma GH response to BM pretreatment in control rats and those with lesions of the arcuate nucleus indicates a hypothalamic action of glucocorticoids, presumably on somatostatin and GHRH neurones. In control animals the effects appear to be counterbalancing, but following destruction of GHRH neurones an uncompensated inhibitory influence was observed. Male MSG-treated rats had lower body weight (-25%) and reduced hypothalamic GHRH (-89%) and pituitary GH content (-69%) compared to male controls. Female rats which had undergone the same neonatal MSG treatment (n = 40) when sacrificed 1 week after their male counterparts showed similar reductions in body weight (-15%), hypothalamic GHRH (-74%), and pituitary GH (-67%).(ABSTRACT TRUNCATED AT 250 WORDS)     

Effect of testosterone on growth hormone secretion in female rats during a continuous infusion of growth hormone releasing factor

The effect of testosterone on growth hormone (GH) secretory pattern during a 6-hour continuous infusion of human GH-releasing factor (GRF) (1-44) NH2 was observed in unrestrained adult female Wistar rats. Rats had been ovariectomized or sham operated 6 weeks previously. Three weeks after the ovariectomy, the rats received sesame oil or testosterone propionate at a dose of 1 or 2 mg s.c. daily for 21 days. All rats were provided with two indwelling cannulae: one in the right atrium for undisturbed blood collection and the other in the inferior vena cava for vehicle or GRF infusion. Vehicle or GRF was administered by an infusion pump at a dose of 50 ng/kg/min for 6 h. Serial blood specimens were obtained every 20 min. Sham-operated adult female Wistar rats exhibited a high-frequency, low-amplitude pulsatile GH secretion during a 6-hour vehicle infusion. When they received a 6-hour continuous infusion of GRF, the amplitudes of GH pulses and baseline GH values were markedly augmented, but the pulse frequency remained unaltered. The GH secretory pattern during a 6-hour vehicle infusion among ovariectomized rats was similar to that of sham-operated female rats, whereas the magnitude of elevation of GH pulse and baseline level in ovariectomized rats were significantly lower than in sham-operated rats. The ovariectomized female rats that had received 2 mg testosterone for 21 days showed a low-frequency, regularly timed, high-amplitude pulsatile GH secretion, and GH values during the intervening period were low. This GH secretory pattern was indistinguishable from that in adult male rats.(ABSTRACT TRUNCATED AT 250 WORDS)     

Post-somatostatin rebound secretion of growth hormone is dependent on growth hormone-releasing factor in unrestrained female rats

To examine the characteristics of GH secretion following the termination of the infusion of somatostatin, unrestrained adult female Wistar rats were subjected to repeated infusions of somatostatin separated by 30-min control periods. When somatostatin was infused for 150 min at a dose of 3, 30 or 300 micrograms/kg body wt per h, the magnitude of the rebound GH secretion increased in a dose-dependent manner. The infusion of somatostatin at a dose of 300 micrograms/kg body wt per h for 60, 150 or 240 min progressively augmented the size of the rebound GH secretion. When an antiserum to rat GH-releasing factor (GRF) was injected i.v. 10 min before the end of the infusion, the peak amplitude of the rebound GH secretion (300 micrograms/kg body wt, 150 min) was reduced to less than 20% of that of control rats. The rebound GH secretion (300 micrograms/kg body wt per h, 150 min) was augmented by a bolus injection of human GRF (1 microgram/kg body wt). The combined effect of the end of infusion of somatostatin and a bolus injection of GRF on the amount of GH secreted was additive. The plasma GH response to GRF was completely inhibited when human GRF (3 micrograms/kg body wt per h) and somatostatin (300 micrograms/kg body wt per h) were infused simultaneously for 150 min. The magnitude of the rebound GH secretion following the termination of the co-administration was larger than that following the somatostatin infusion alone, but this rebound was not enhanced by a bolus injection of human GRF.(ABSTRACT TRUNCATED AT 250 WORDS)     

Somatostatin inhibition of growth hormone secretion by somatotropes from male, female, and androgen receptor-deficient rats: evidence for differing sensitivities

To investigate the role of somatostatin (SRIF) in regulating sexually dimorphic GH secretion, we used a reverse hemolytic plaque assay and acutely dispersed somatotropes from age-matched normal male, normal female, and androgen receptor-deficient, testicular feminized (Tfm) rats. Hemolytic plaques were developed after a 90-min incubation in the presence of GH antiserum, 10 nM GH-releasing hormone (GHRH), and the following concentrations of SRIF: 0, 0.001, 0.003, 0.01, 0.03, 0.1, 0.3, 1, 3, 10, 30, and 100 nM. Additional studies were performed with 0 or 100 nM SRIF in the absence of GHRH. The absolute number of somatotropes (x10(6); mean +/- SEM) recovered from the pituitaries of Tfm rats (1.73 +/- 0.18) was significantly greater than that from the males (1.11 +/- 0.13; P = 0.01); the number from female rats (1.30 +/- 0.15) was not different from that of either male or Tfm animals. GHRH-stimulated GH secretion, as estimated by the mean GH plaque area (micron2 x 10(4); mean +/- SEM) in the absence of SRIF, was greater for somatotropes from male rats (3.36 +/- 0.41) than that for either Tfm (2.27 +/- 0.32; P = 0.02) or female (1.78 +/- 0.24; P = 0.001) rats; values for the latter two groups did not differ. However, mean GH plaque areas for each group during maximal SRIF inhibition in either the presence or absence of GHRH were indistinguishable from each other and from mean plaque areas obtained under basal conditions. As demonstrated by a lesser EC50 value (0.04 +/- 0.02 nM; mean +/- SEM), somatotropes from female rats were more sensitive to the inhibitory effect of SRIF than were those from either male (EC50 = 1.82 +/- 0.45; P = 0.0001) or Tfm (EC50 = 0.74 +/- 0.22, P = 0.0001) rats; values for the latter two groups were indistinguishable. These observed differences suggest that gender and/or the gonadal hormone environment may be important determinants of the inhibitory effects of SRIF on GH secretion by the somatotrope. While these gender-associated differences may represent effects of the gonadal hormones directly on the somatotrope, they could reflect modulation of the secretion of hypothalamic SRIF and/or GHRH by the prevailing gonadal hormone environment. Such gender-related differences may contribute to the overall sex-dependent patterns of GH secretion in the intact animal.     

Inhibitory effect of neurotensin on gastric acid secretion in rats

Neurotensin has been shown to inhibit gastric acid secretion when administered in pharmacological doses, but no information has been available concerning its possible dose-related effect during intravenous infusion. In this study, a dose-related and reversible inhibitory effect of neurotensin was demonstrated in pentobarbital-anesthetized female Sprague-Dawley rats. The rats underwent continuous gastric perfusion with saline, 1 ml/min, and intravenous infusion of both pentagastrin and neurotensin. Inhibition of acid secretion did not depend upon the occurrence of hypotension, and ranged from 35±7% of maximal acid output at 0.24 nmol/kg/hr to 60 ±10% at 7.2 nmol/kg/hr of neurotensin. Blood levels of C-terminal neurotensin-like immunoreactivity were proportional to the dose of peptide infused and were 52 fmol/ml during infusion of 0.24 nmol/kg/hr, a dose that significantly inhibited pentagastrin-induced acid secretion. Thus, a model has been developed to study the effect of neurotensin infusion on acid secretion; the concentration of plasma neurotensin-like immunoreactivity at which inhibition occurs in this model is similar to the concentration reported to occur after a nutrient stimulus.This work was supported by Merit Review funds from the Veterans Administration.     

Effect of androgen on growth hormone secretion and growth in boys with short stature.

The response of plasma growth hormone (GH) to insulin-induced hypoglycaemia (IIH) and arginine infusion (AI) was studied in 22 young males (ages 8 to 17 years) with short atature and absent or delayed sexual maturation, before and after androgen administration. During initial evaluation, 5 patients had blunted GH response to IIH, 12 responded subnormally to AI and 4 failed to respond normally to either stimulus. These same studies were repeated in each patient following androgen administration. The source of androgen was as follows: a) 5 days of testosterone propionate (25 mg intramuscularly daily) in 20 patients. b) Methyltestosterone, 10 mg t.i.d. orally for four days in the other 2 subjects. In almost every case, androgen administration resulted in raising the levels of fasting GH and enhancement of the GH responses to IIH and AI was observed. Patients manifesting subnormal GH responses to these stimuli before androgen consistently demonstrated a normal response when challenged with identical stimuli during androgen administration. Growth velocities during the year following these studies were significantly increased in most instances and the growth spurts correlated well with the progression of sexual maturation. Sustained improvement in the GH responses to IIH and AI were uniformly observed in 3 patients when repetitive studies were performed 8 to 12 months later during spontaneous advancing sexual development. The results indicate that brief androgen administration can be helpful in delineating the cause of growth retardation in boys with short stature and delayed sexual maturation, particularly when the diagnosis of isolated growth hormone deficiency is suspected. They also offer prognostic value in determining growth potential in this same group of young males.     

Cross-talk between excitatory and inhibitory amino acids in the regulation of growth hormone secretion in neonatal rats

Experiments were carried out in 5-day-old male and female rats in order to assess the respective roles of distinct gamma-aminobutyric acid (GABA) receptor subtypes in the control of growth hormone (GH) secretion in the newborn rat and to identify the potential sexual dimorphism of GABA actions on GH release. The interplay of GABA and excitatory acids (EAAs) to control GH secretion was also analysed. Effects of specific GABA(A), GABA(B) and GABA(C) agonists upon GH secretion as well as of co-administration of GABA and agonists and antagonists of EAA receptors were monitored in 5-day-old male rats. The data confirm that GABA stimulates GH secretion in 5-day-old rats, but indicate that the effect is sexually dimorphic, being greater in females. Stimulation of GH secretion by GABA is mainly mediated by GABA(A) receptors, while GABA(C) appears to be ineffective. Stimulatory effects of GH by GABA and muscimol, a GABA(A) receptor agonist, are blocked by pretreatment with (+)-5-methyl-10,11-dihydro-5H-dibenzo[a,d]cyclophepten-5-10-imine hydrogen maleate (MK-801), an antagonist of N-methyl-D-aspartate (NMDA) receptors, but not by 1,2,3,4-tetrahydro-6-nitro-2,3-dioxo-benzo[f]-quinoxaline-7-sulfonamide (NBQX), an antagonist of (+/-)-alpha-amino-3-hydroxy-5-methylisoxazole-4-propionic acid (AMPA) receptors. Simultaneous administration of GABA and NMDA was additive on GH release while basal GH secretion was not affected by either bicuculline or the respective blockers of GABA(A) and GABA(B) receptors. In conclusion, our results suggest that cross-talk between inhibitory (GABA) and EAAs is involved in the control of GH secretion in neonates.     

Effect of the serotoninergic system on luteinizing hormone secretion in prepubertal female rats

The effect of 5-hydroxytryptophan (5-HTP), a serotonin precursor, and p-chloroamphetamine (PCA), a serotonin neurotoxin, selective for serotoninergic neurons, that depletes serotonin (5-HT) levels in brain, on the luteinizing hormone (LH) release response to estrogen-progesterone (E-P) was studied in prepubertal female rats of different ages. E-P decreased LH levels on days 16, 18 and 20, increasing the levels of the pituitary hormone at day 26 of age. Destruction of the serotoninergic system advanced the onset of the positive feed-back mechanism, since the rats pretreated with PCA showed at day 20 an LH release by E-P administration while in the controls of the same age the ovarian hormones decreased the LH concentration. On the other hand, PCA potentiated the positive feed-back mechanism of E-P on LH in 26-day-old rats, while at this age the LH release response to E-P was significantly reduced by the administration of 5-HTP. These results suggest that the serotoninergic system has an inhibitory effect on the development of the positive feed-back of ovarian steroids on LH secretion, that could be representative of a regulatory participation of serotonin in the onset of puberty. 5-HTP stimulated LH release on days 16, 18 and 20, but did not modify the LH concentration of day 26. Since between 20 and 26 days of age the positive feed-back mechanism matures, the possibility arise that the modification in the effect of serotonin on LH release on day 26 is connected with the physiological changes in the gonadotropin control that occur after day 20 in the female rat.     

Pulsatile growth hormone secretion in the ovine fetus and neonatal lamb

The ontogenesis of the pulsatility of GH in the ovine fetus was determined by obtaining blood samples at 20-min intervals for 3-h periods from fetuses (n = 33) at various stages of development (76-147 days gestation), and in neonatal life (n = 19). A significant increase (P less than 0.01) in the GH mean, nadir and maximum, and pulse height was observed between the ages of 100 and 130-139 days of gestation. An analysis of the difference in the mean, maximum and nadir concentrations between 100 and 139 days of gestation revealed that males had higher GH levels than females (P less than 0.05). There was a significant fall in plasma GH concentrations from 140 days of gestation to term, but before the onset of active labour. There was a more rapid fall in the circulating levels of fetal GH directly following birth. Immediately before birth fetal GH levels were still relatively high, but within 60 min of birth they had fallen by more than 80%. It is suggested that these changes in the pulsatile pattern of GH release are a consequence of both maturational changes in the hypothalamic-pituitary unit and the effects of pregnancy-related factors on GH release. The sexually dimorphic nature of GH release in the adult is also observed in the sheep fetus during late gestation.     

Inhibitory action of the alpha 1-adrenergic receptor on growth hormone secretion in the dog

Intravenous administration of methoxamine ( METHOX , 5 mg, iv), a specific alpha 1-adrenergic agonist, reduced baseline GH levels of four unanesthetized beagle dogs 45 and 60 min post-injection and completely abolished the GH-releasing effect of the alpha 2-adrenergic agonist clonidine ( CLON , 4/ micrograms/kg, iv). Prazosin (PRA, 0.1 mg/kg, iv), an alpha 1-adrenergic antagonist, administered before METHOX re-instituted the GH-releasing effect of CLON . METHOX administered at the starting of an arginine infusion (ARG, 10% solution, 3.3 ml/min X 30 min) reduced consistently the GH releasing effect of the latter while, conversely, pretreatment with PRA, strikingly potentiated the GH-releasing effect of the amino acid (2 dogs). METHOX did not alter the GH-releasing effect of human pancreatic GH-releasing factor (hpGRF-40, 1/microgram/kg, iv), though it consistently delayed the occurrence of the GH secretory peak following hpGRF-40. These data indicate that alpha 1-adrenergic receptors located in the central nervous system, inhibit in the dog tonic and stimulated GH secretion.