The effects of a continuous infusion of hexarelin on pulsatile growth hormone release, growth axis and galanin gene expression and on the response of the growth axis to growth hormone-releasing hormone.

The effect of a 6 hour continuous infusion of Hexarelin (100 micrograms/hour) on GH peak frequency, amplitude and duration, GH trough concentrations, the interval between successive peaks and the pituitary responsiveness to GHRH, as well as GH axis and galanin mRNA contents, were examined in conscious adult male rats. Plasma GH concentrations peaked within 15 minutes after the initiation of Hexarelin infusion, but returned to baseline levels by 60 minutes. No significant differences between Hexarelin and saline infused rats were noted for any of the parameters of pulsatile GH release analyzed. However, following a 6 hour infusion, rats treated with Hexarelin demonstrated a greater GH responsiveness to GHRH (delta GH: 57 +/- 16 ng/ml for Hexarelin infused; 21 +/- 7 ng/ml for saline infused; p < 0.05). Furthermore, the rats infused with Hexarelin demonstrated decreased GHRH and increased hypothalamic galanin mRNA contents as compared to the saline infused rats, while hypothalamic somatostatin and pituitary GH mRNA contents appeared unchanged. Rats infused with Hexarelin had lower pituitary galanin mRNA content than did the rats which were infused with saline. Collectively, these results suggest that Hexarelin may not act via alteration of somatostatin synthesis and that suppression of somatostatin's action at the pituitary can not be excluded. The current study also suggests that other hypothalamic pathways aside from those currently defined for the growth axis may be involved in the mechanism by which Hexarelin and the other GH-releasing peptides elicit GH release.

Acute and chronic galanin administration decreases hypothalamic galanin synthesis in both male and female adult rats: evidence for a long-loop galanin autofeedback.

The secretion of growth hormone (GH) in both male and female rats is controlled by two main neuropeptides, GH-releasing hormone (GHRH), which is stimulatory, and somatostatin, which is inhibitory. Recently, it has been shown that galanin (GAL) also stimulates GH secretion, although the underlying mechanism is still unknown. It was the aim of this study to begin to elucidate if and how GAL regulates its own production at the hypothalamic and pituitary level. Rats underwent the following experimental trials. In experiment 1, adult male and female rats had blood samples collected at -15 minutes, -7.5 minutes, and immediately preceding a subcutaneous (s.c.) injection of GAL at a dose of either 50 or 200 microg/kg. Blood samples were collected at 5, 10, 15, 30, and 60 minutes, and the GH concentration was measured using a radioimmunoassay. The tissues were collected and analyzed for mRNA levels of hypothalamic and pituitary GAL. In experiment 2, adult male and female rats were treated long-term with 200 microg/kg GAL for 7 days s.c., and the pituitary and hypothalamus were analyzed for GAL mRNA. Serum GH concentrations were significantly increased in acutely dosed male and female rats regardless of the dosage level. For the male and female animals acutely dosed with both 50 and 200 microg/kg GAL, hypothalamic GAL mRNA was decreased, whereas pituitary GAL mRNA was affected by 200 microg/kg GAL only in females (increased). For the animals treated long-term with GAL, hypothalamic GAL mRNA was decreased while mRNA for pituitary GAL was increased. We conclude that regardless of the dosage and duration of treatment, administration of GAL negatively regulates hypothalamic GAL mRNA in a non-gender-specific way. Pituitary GAL synthesis appears to be stimulated particularly during chronic SCGAL administration.

Short-term glucocorticoid administration decreases both hypothalamic and pituitary galanin synthesis in the adult male rat.

Galanin (GAL) is a peptide that has been implicated in the regulation of the growth axis. It is generally accepted that GAL can increase serum growth hormone (GH) levels, although the underlying mechanism for this increase is unknown. It is well known that long-term glucocorticoid treatment alters in vivo GH secretion, since there is a decrease in serum GH in response to stimuli. It has previously been shown in our laboratory that administration of GAL can overcome the effects of glucocorticoid administration on GH secretion. The aim of the present study was to determine the effects of long-term glucocorticoid administration on the regulation of hypothalamic and pituitary GAL mRNA levels. Adult male rats were treated for 72 hours with the synthetic glucocorticoid dexamethasone ([DEX] 40 microg/kg/d intraperitoneal injections). RNase protection assays were performed on both the hypothalamus and pituitary for the presence of GAL mRNA. As expected, DEX significantly decreased somatic growth, as evidenced by a decrease (50%) in the weight gain of glucocorticoid-treated versus control animals. It was also demonstrated that in both the hypothalamus and pituitary, glucocorticoid treatment reduced the level of GAL mRNA (to 11% and 6.5%, respectively) compared with the control condition. We conclude that the decrease in GAL mRNA may lead to a decrease in GAL secretion, which in turn may be involved in the glucocorticoid-induced inhibition of GH secretion.

Effects of food deprivation on the GH axis: immunocytochemical and molecular analysis.

Neuroendocrine mechanisms governing growth hormone (GH) secretion are sensitive to nutritional status since the normal pulsatile pattern of GH release is disrupted during conditions of food deprivation or malnutrition. A reasonable hypothesis for this occurrence is the alteration of somatostatin and GH-releasing hormone (GHRH) synthesis, storage and secretion. In this study, we investigated the effects of food deprivation on GH, GHRH, hypothalamic and pituitary galanin (GAL), and somatostatin through immunocytochemical and mRNA analysis. Adult male rats were subjected to 72 h of food deprivation, during which an average of 18% total body weight was lost. ICC studies were performed on brain sections from the rostral, middle and caudal regions of the median eminence of the hypothalamus using the avidin-biotin-peroxidase method. Immunocytochemical results were generated for the percent area and optical density (intensity) of immunostaining in the median eminence. Messenger RNA analyses were performed using sense and antisense riboprobes produced from cDNA clones for GH, GHRH, somatostatin and GAL. Food deprivation decreased somatostatin immunostaining in middle and caudal regions of the median eminence; similarly, food deprivation resulted in decreased GHRH immunostaining in rostral and middle sections of the median eminence of the hypothalamus. mRNA levels for somatostatin, GHRH and GH and GAL were also reduced by food deprivation. Our data suggest that suppressed GH secretion in food-deprived rats may reflect a general downregulation of the neuroendocrine and pituitary GH axis.

Immunocytochemical and molecular analysis of the effects of glucocorticoid treatment on the hypothalamic-somatotropic axis in the rat.

Glucocorticoids are potent inhibitors of linear growth and growth hormone (GH) secretion when secreted or administered in pharmacological amounts in vivo. The mechanisms involved require further clarification although enhanced somatostatin tone has been suggested to play a role. In this study, we investigated the effects of excess glucocorticoids on pituitary GH, hypothalamic GHRH and hypothalamic somatostatin through immunocytochemical (ICC) and mRNA analysis. Twelve adult male rats were injected daily with dexamethasone (40 micrograms/day, i.p.) or saline for 4 days. ICC studies were performed on brain sections from the rostral, middle and caudal regions of the median eminence of the hypothalamus using the avidin-biotin-peroxidase method. Messenger RNA analyses were performed using sense and antisense riboprobes produced from GH, GHRH and somatostatin cDNAs. Immunocytochemical results were generated for the percent area and intensity (optical density) of immunostaining in the median eminence. Glucocorticoids increased somatostatin immunostaining of the rostral, middle and caudal regions of the median eminence while GHRH staining was only reduced in the rostral region of the median eminence and unchanged in the other hypothalamic regions. GH and somatostatin mRNA levels dramatically increased following glucocorticoid treatment concomitantly with a decrease in GHRH mRNA levels. Our data suggest that increased somatostatin synthesis and storage and a decrease in GHRH mRNA synthesis play a major role in the GH inhibitory effects of glucocorticoids.

Hypothalamic control of growth hormone (GH) secretion in type I diabetic men: effect of the combined administration of GH-releasing hormone and hexarelin, a novel GHRP-6 analog.

Insulin dependent (type I) diabetic patients show abnormal growth hormone (GH) secretion. Hexarelin is an analog of GHRP-6 which releases GH in part via somatostatin inhibition. The aim of our study was to evaluate the effects of hexarelin and GHRH, administered either alone or in combination, on GH secretion in 10 type I diabetic and 7 normal men. All the subjects were administered: 1) human GHRH (1-29) NH2 100 micrograms i.v. bolus at 0 min; 2) hexarelin 100 micrograms i.v. bolus at 0 min; 3) hexarelin 100 micrograms + hGHRH 100 micrograms i.v. bolus at 0 min. In type I diabetic patients significantly greater GH responses to GHRH and hexarelin have been observed with normal subjects. Hexarelin caused a significantly (p < 0.05) greater GH response as compared to GHRH in both diabetic and control subjects. After the administration of hexarelin+GHRH, a significant increase in both GH absolute and peak levels as compared to hexarelin or GHRH alone was found in all the subjects. However, the GH responses to the combined stimuli were not significantly different in diabetics as compared to normals; moreover, the interaction of GHRH and hexarelin was synergistic in controls and additive in diabetics. We hypothesize that a reduction in the hypothalamic somatostatin inhibitory tone combined with increased pituitary GH production may be responsible for the pattern of the GH responses to hexarelin and GHRH observed in our type I diabetic patients.

Influence of thyroid hormones on the regulation of growth hormone secretion.

The effects of thyroid hormones on GH secretion and the mechanisms underlying their action are very similar in man and the laboratory animal. We feel that it is possible to organize the available data into a unique pathophysiological model explaining these complex interactions (Table 1). In summary, physiological levels of circulating thyroid hormones are necessary to maintain normal pituitary GH secretion owing to their direct stimulatory actions. When the serum concentrations of thyroid hormone increase above the normal range there is an increase in hypothalamic somatostatin tone, which in turn suppresses pituitary GH secretion and overrides any stimulatory effects. The suppression of GH secretion by thyroid hormones may be mediated at the hypothalamic level also by a decrease in GHRH release.

Adrenergic and cholinergic involvement in basal and growth hormone-releasing hormone-stimulated growth hormone secretion in glucocorticoid-treated rats.

Glucocorticoids are known to inhibit GH secretion via somatostatin. The aim of our study was to elucidate the involvement of somatostatin in the GH-releasing action of the alpha 2 agonist clonidine and the cholinergic agent pyridostigmine in conscious, freely-moving rats chronically treated with dexamethasone. After seven days of chronic glucocorticoid treatment, animals received an i.v. injection of either saline (1 ml/kg) or clonidine (150 micrograms/kg) or pyridostigmine (100 micrograms/kg) at -15 min. Three blood samples were then drawn (-10 min, -5 min, and 0 min) to assess the GH response to either clonidine or pyridostigmine alone. After the 0 min sample, saline (1 ml/kg) or GNRH (500 ng/kg) was injected i.v. and additional blood samples were drawn from 5 to 30 min. The GH response to clonidine alone or combined with GNRH in rats treated with dexamethasone was significantly lower (p < 0.05) as compared to vehicle-treated rats. The GH response to pyridostigmine alone or combined with GNRH did not significantly differ between vehicle- and dexamethasone-treated rats. These data suggest that in the rat the mechanism of action of clonidine is mainly to stimulate endogenous GNRH secretion, while pyridostigmine appears to predominantly act by decreasing hypothalamic somatostatin.

Changes in the growth hormone axis due to exercise training in male and female rats: secretory and molecular responses.

GH secretion is altered by exercise in humans. In an attempt to investigate the underlying mechanisms, we developed a rodent model. GH secretion was assayed in male and female rats that were sedentary (not exercised), acutely exercised, and chronically exercised. Sedentary males showed typical pulsatile GH secretion. The acutely exercised males had low GH concentrations during the exercise bout, but showed partial recovery of GH pulses during the 5.5-h postexercise period. GH secretion in the chronically exercised males was low during both the exercise and postexercise periods. Sedentary females displayed the typical pattern of GH secretion for this sex. The acutely exercised females had low GH concentrations during the exercise period; the pulsatile pattern of GH secretion did not return during the postexercise period. In contrast, the chronically exercising females had suppressed GH secretion during the exercise bout, but concentrations immediately returned to normal during the postexercise bout. The effects of exercise on GH, GH-releasing hormone (GHRH), and somatostatin messenger RNA (mRNA) levels using Northern and slot blot analyses were also determined. Acutely and chronically exercised male rats had decreased levels of GH mRNA compared to sedentary male rats. The acutely exercised female rats had increased levels of GH mRNA compared to the sedentary females, whereas the chronically exercised females had decreased levels. GHRH mRNA levels in acutely exercising male rats was decreased and in chronically exercising male rats was increased compared to those in the sedentary controls. The pattern of GHRH mRNA in female rats was the opposite of this. Somatostatin mRNA levels decreased in acutely exercised male rats and were not affected in chronically exercised male rats. This signal increased in both acute and chronically exercised female rats. These studies suggest that GH secretion is suppressed in response to exercise in the rat. This contrasts with the increase observed after exercise in humans.

Long-term changes of somatotrophic function induced by deprivation of growth hormone-releasing hormone during the fetal life of the rat.

We have studied the effects of intra-amniotic administration of an anti-GH-releasing hormone serum (GHRH-Ab) on day 16 of fetal life in the rat, when the ontogenetic development of the GHRH neuronal system occurs. Control animals received normal rabbit serum. Following delivery, body weight was monitored for the next 30 days as an index of somatic growth, and the following indices of somatotrophic function were determined: plasma and pituitary GH, pituitary GH mRNA, hypothalamic GHRH and somatostatin mRNA, and the in vivo GH responsiveness to GHRH. At birth, GHRH-Ab-treated rats had a body weight that was equivalent to that of control rats but, starting from postnatal day 6 up to day 30, they had a significantly reduced body weight. Pituitary weight, the absolute pituitary GH content and GH mRNA levels were lower in experimental compared with control rats, while pituitary GH concentrations were similar in the two groups, thus implying that there was a defect, not only in GH synthesis, but also in GH release. In agreement with this theory, basal GH levels and GHRH-stimulated GH secretion were reduced in GHRH-Ab-treated rats but, in contrast, hypothalamic regulation of GH secretion appeared to be working in these rats as they were still able to respond to the low plasma GH by increasing GHRH and decreasing somatostatin mRNA levels. These findings indicate that deprivation of GHRH during fetal life induces long-lasting changes of growth rate and somatotrophic function.(ABSTRACT TRUNCATED AT 250 WORDS)

Hypothalamic regulation of growth hormone secretion during food deprivation in the rat.

Suppressed pulsatile GH secretion in food-deprived rats has been hypothesized to be due to an increase in hypothalamic somatostatin secretion. We investigated this hypothesis and the role of GHRH in regulating GH secretion during food deprivation using two different models. In experiment one, rats were food deprived for 72h during which time they received a saline infusion (n = 5). At the same time rats were normal fed for 72h during which time they received a somatostatin infusion (5 micrograms/h, n = 7). After the 72h infusion period, all rats received two iv injections of GHRH (1 microgram/rat) at 2h intervals. GH concentrations in food-deprived rats rose from approximately 10 ng/ml to 400-800 ng/ml in response to both GHRH injections. This increase was significantly greater (p < 0.01) than the GH response (100-400 ng/ml) observed in somatostatin-infused animals. The significantly higher GH response observed in food-deprived rats as compared to somatostatin-infused, normal-fed rats suggests that somatostatin concentrations may decrease during food deprivation. In experiment two, rats were infused for 5h with either saline (n = 6) or GHRH (10 micrograms/h, n = 9) at the end of a 72h fast. GH concentrations did not change in saline-infused animals. In contrast, GH concentrations significantly increased (p < 0.01) upon initiation of the continuous GHRH infusion. Yet, this release of GH was pulsatile in nature. Pulsatile GH secretion in the presence of a constant GHRH infusion suggests that pulsatile somatostatin release from the hypothalamus is maintained during food deprivation. These studies suggest that during food deprivation in the rat 1) absolute concentrations of somatostatin decrease, but its pattern of secretion remains pulsatile, and 2) decreased GHRH release may be responsible for the absence of spontaneous GH pulses.

Basic counterpoint: mechanisms and pathways of gonadal steroid modulation of growth hormone secretion.

In spite of the different patterns of GH secretion observed in male and female rats, it can be argued that there are limited differences between the mechanism of action of androgens and estrogens as reported in the literature. However, we feel that it is possible to organize the available data into a unique physiological model explaining the sex-based differences in GH secretion in the rat. Thus, it can be proposed that the greater spontaneous GH peaks observed in the male with respect to the female rat may be due to an androgen-mediated enhancement of both GHRH secretion at the hypothalamic level and GH secretion at the pituitary level. The lower GH troughs observed in the male as compared to the female rat may be due to increased interpeak somatostatin secretion induced by the androgens with respect to the estrogens. It is likely that these high GH peaks and low GH troughs establish a recycling mechanism through established feedback mechanisms. That is, the high GH peak, induced by GHRH, stimulates somatostatin secretion such that a very low GH trough follows. In turn, this low GH trough, in the high somatostatin environment, establishes the correct neuroendocrine milieu for the next high GH peak, and so on. Additional studies will help clarify this model and hopefully provide a better understanding of the mechanisms regulating the interaction between gonadal steroids and GH.

Hypothalamic regulation of impaired growth hormone secretion in diabetic rats. 1. Studies in streptozotocin-induced diabetic rats.

Growth hormone (GH) secretion is blunted in diabetic rats. In the present experiment we observed that pituitary GH concentrations and the plasma GH response to an exogenous dose of growth hormone-releasing hormone (GHRH) is decreased in streptozotocin-induced diabetic rats (p less than 0.02) with respect to normal rats. In an attempt to determine if increased somatostatin (SRIF) secretion is responsible for the decreased GH secretion, we studied the effect of modulating SRIF tone on the GH response to GHRH in normal and streptozotocin-induced diabetic rats. Rats were pretreated with either normal sheep serum and saline (NSS+SAL), somatostatin antibodies (SRIF-Ab), or pyridostigmine (PD), an acetylcholinesterase inhibitor hypothesized to reduce hypothalamic SRIF secretion. Pretreatment of normal rats with SRIF-Ab or PD resulted in an increased GH response to exogenous GHRH in comparison to NSS+SAL-pretreated normal rats at 5 min postinjection. In contrast, pretreatment of diabetic rats with SRIF-Ab or PD did not alter the GH response to exogenous GHRH when compared to NSS+SAL-pretreated diabetic animals. These results suggest that the blunted GH response to exogenous GHRH observed in streptozotocin-induced diabetic rats may not be due to an increase of endogenous SRIF tone.

Hypothalamic regulation of impaired growth hormone secretion in diabetic rats. 2. Studies in spontaneously diabetic BB Worcester rats.

In the present study we investigated the effects of modulating endogenous somatostatin (SRIF) on the GH response to growth hormone-releasing hormone (GHRH) in spontaneously diabetic BB/Wor rats and nondiabetic littermates. Plasma growth hormone (GH) concentrations following injection of GHRH (500 ng/kg, i.v.) were measured in the rats after pretreatment with either normal sheep serum+saline (NSS+SAL), somatostatin antibody (SRIF-Ab), or pyridostigmine bromide (PD), an acetylcholine esterase inhibitor hypothesized to decrease hypothalamic SRIF tone. The GH response to GHRH in spontaneous diabetic rats pretreated with NSS+SAL was significantly lower (p less than 0.05) than the response observed in the nondiabetic group. SRIF-Ab pretreatment reversed the blunted GH response observed in the diabetic rats. However, PD pretreatment was not effective. These results indicate that the blunted GH response observed in BB/Wor diabetic rats is reversed by neutralization of endogenous SRIF with SRIF-Ab and leads to the conclusion that SRIF plays an active role in modulating GH secretion in spontaneously diabetic rats. The failure of PD to modulate the GH response suggests this acetylcholine agonist is ineffective in this animal paradigm.

Effects of pyridostigmine on spontaneous and growth hormone-releasing hormone stimulated growth hormone secretion in children on daily glucocorticoid therapy after liver transplantation.

OBJECTIVES: We aimed to investigate both nocturnal spontaneous and morning growth hormone (GH)-releasing hormone (GHRH)-induced GH secretion in children on daily glucocorticoid treatment after liver transplantation and to evaluate the effect of pyridostigmine (an acetylcholinesterase inhibitor thought to reduce hypothalamic somatostatin tone) on GH secretion in these patients. DESIGN: We performed a randomized, single-blind, cross-over study. PATIENTS: We studied three male and three female juvenile patients, within a year of orthotopic liver transplantation and under immunosuppressive glucocorticoid therapy (mean dose +/- SEM, 5.92 +/- 0.63 mg/day) and five normal children (four males, one female). MEASUREMENTS: Both nocturnal spontaneous and morning GHRH-induced GH secretion were evaluated after administration of placebo, 1 tablet p.o., or pyridostigmine, 2 mg/kg p.o. RESULTS: Spontaneous GH. Placebo: in liver transplanted children nocturnal GH secretion (mean GH level 10.8 +/- 2.0 mU/l) was not significantly different with respect to normal children (mean GH level 12.8 +/- 1.2 mU/l); pyridostigmine: nocturnal GH secretion was significantly increased as compared to placebo in subjects with liver transplantation but not in normal children. GHRH test. Placebo: liver transplanted patients showed a blunted GH response to GHRH with respect to normal children; pyridostigmine: the GH responses to GHRH (P less than 0.05) increased as compared to placebo and did not differ significantly in the two groups. CONCLUSIONS: Our data suggest a steroid-mediated increase in hypothalamic somatostatin tone in liver transplanted children.

Deprivation of growth hormone-releasing hormone early in the rat's neonatal life permanently affects somatotropic function.

This work investigated in rats whether passive immunization against the endogenous GHRF in the early postnatal period led to permanent alterations of somatotropic function, similar to those observed in several human growth disorders, e.g. constitutional growth delay (CGD). On postnatal days 1, 2, 4, 6, 8, and 10, rats were given an anti-GHRF-serum (GHRH-Ab, 100 microliters/rat, sc) and were tested 1, 30, and 60 days after this treatment for basal and GHRH-stimulated GH secretion both in vivo and in vitro. GHRH-Ab reduced both basal and GHRF-stimulated GH secretion at all intervals and induced marked and chronic impairment of growth rate. The following differences were observed in the GHRH-Ab treated rats compared to normal rabbit serum-treated controls: 1) GH biosynthesis (incorporation of L-[3H]leucine into the electrophoretic band of GH): reduction of about 70%, 1 day but not 30 days after treatment; 2) Pituitary weight: significant reduction in absolute weight (30-40%) at all posttreatment intervals, and relative weight, 1 and 30 days after treatment. 3) Pituitary GH concentration: significant reduction in GH content (about 40%) but not concentration, at all posttreatment intervals; 4) Percentage of somatotrophs (immunocytochemistry): about 40% reduction 1 day, but not 30 and 60 days after treatment; 5) Hypothalamic somatostatin messenger RNA (mRNA) levels in situ hybridization): selective reduction (40%) in the periventricular nucleus 1 day but not 30 days after treatment; 6) Hypothalamic somatostatin cell number (immunocytochemistry): no significant changes in any hypothalamic area at any interval; 7) Pituitary somatostatin binding (in situ autoradiography): significant reduction, 1 day and 30 days after treatment; 8) Somatostatin inhibition of GH release "in vitro": somatostatin effect on GH release was reduced 30 days after treatment. These and previous data indicate that: 1) Transient deprivation of GHRF in the immediate postnatal period of the rat leads to permanent impairment of growth rate and somatotropic function; 2) GHRF deficiency itself or through reduction of GH secretion impairs somatostatin functions temporarily in the hypothalamus and permanently in the pituitary; 3) This rat model may mimic some forms of growth disorders in humans and holds promise as useful tools for investigating the underlying pathophysiological mechanisms.

Pyridostigmine blocks the inhibitory effect of glucocorticoids on growth hormone-releasing hormone stimulated growth hormone secretion in normal man.

Glucocorticoids have been shown to inhibit GH secretion in normal man when acutely and chronically administered in pharmacological amounts. Pyridostigmine (PD), an acetylcholinesterase inhibitor, is able to elicit GH secretion when administered alone and to enhance the GH response to GHRH in normal subjects probably via a decrease in the hypothalamic release of somatostatin. The aim of the present study was to investigate the influence of glucocorticoids on the GH response to PD administered either alone or in combination with GHRH in normal adult subjects. Six healthy adult volunteers underwent six experimental protocols. They received 1) human (h) GHRH(1-29)NH2, 100 micrograms injected as an iv bolus; 2) cortisone acetate, 50 mg administered orally (po) 60 min before an hGHRH iv bolus injection; 3) PD, 120 mg administered po, 60 min before an hGHRH iv bolus injection; 4) PD and cortisone acetate, administered po 60 min before an hGHRH iv bolus injection; 5) PD, administered po 60 min before a saline iv bolus injection; 6) PD and cortisone acetate administered po 60 min before a saline iv bolus injection. Mean GH levels, peak GH levels, and GH area under the curves (AUCs) were significantly lower after GHRH + cortisone as compared to GHRH alone. However, these parameters were not significantly different after PD + GHRH + cortisone when compared to PD + GHRH and after PD + cortisone when compared to PD alone. We conclude that acute administration of pharmacological amounts of glucocorticoids cannot inhibit the GH response to PD alone or in combination with GHRH. Thus, we hypothesize that the inhibitory action of glucocorticoids on the GH response to GHRH in man may be mediated by an enhancement of either somatostatin release by the hypothalamus or somatostatin action on the pituitary.

Growth hormone (GH) autofeedback action in the neonatal rat: involvement of GH-releasing hormone and somatostatin.

It is known that in adult rats, GH by itself and by promoting secretion of the somatomedins acts at the level of the hypothalamus to trigger release of somatostatin and decrease output of GH-releasing hormone (GHRH), thereby inhibiting further secretion of GH. To assess whether these mechanisms are already operative in the early postnatal period, we have evaluated the effect of short-term administration of GH in 10-day-old rats. Twice-daily s.c. administration of 25 micrograms human GH/rat, from days 5 to 9 of life, significantly reduced pituitary content of GH, decreased hypothalamic levels of GHRH mRNA and abolished the in-vivo GH response to a challenge dose of GHRH (20 ng/100 g body weight, s.c.). GHRH (20 ng/100 g body weight, twice daily, s.c.) given concomitantly with the GH treatment, completely counteracted the inhibitory effect of the latter on pituitary content of GH and restored to normal the in-vivo GH response to the GHRH challenge. These data indicate that impaired secretion of GHRH is involved in the inhibitory effect elicited by GH treatment in infant rats. However, concomitant involvement of hypothalamic somatostatin as a result of GH treatment cannot be ruled out. In fact, pituitaries from rats pretreated with GH responded in the same manner as pituitaries from control rats to the GHRH challenge in vitro.

Neuroendocrine mechanisms regulating growth hormone and prolactin secretion during lactation.

The maternal plasma concentrations of GH and PRL increase dramatically upon the initiation of lactation in the rat. In light of the fact that these two hormones have evolved from one common precursor, we sought to determine if the neuroendocrine mechanisms regulating their concomitant increase during lactation are common or if they are functionally distinct. To evaluate this, lactating rats were passively immunized with antiserum raised against GHRH and then monitored for changes in GH and PRL secretion in response to suckling. On day 9 or 10 postpartum, pups were removed from their mothers at 0800 h. At 1100 h mothers were injected with normal rabbit serum (NRS) or GHRH antiserum (GHRH-ab). At 1400 h a control blood sample was drawn. Pups were then returned to their mothers, with subsequent blood samples drawn over the next 60 min. Plasma concentrations of GH significantly increased to 12.3 +/- 1.0 ng/ml (mean +/- SEM) in NRS-treated females after the return of the pups. In contrast, there was no change in GH concentrations in the females treated with the GHRH-ab. Plasma PRL concentrations rose approximately 200 ng/ml in both the NRS-treated animals and the GHRH-ab-treated ones. Body weight gains of the pups during the 60-min period of lactation were similar in both groups. These results suggest that the neuroendocrine mechanisms regulating the increases in GH and PRL during lactation are distinct and that GHRH is the hypothalamic factor responsible for the increase in GH. Furthermore, these results suggest that acutely interrupting the increase in GH secretion that occurs during lactation does not compromise nursing behavior and performance.

Feed-back effect of growth hormone on hypothalamic opioid and somatocrinin producing neurons.

Reportedly, most acromegalics are refractory to the growth hormone (GH)-releasing effect of central nervous system-acting stimuli. For instance, the synthetic analogue of met-enkephalin (Enk) viz. FK 33-824 fails to alter the high circulating GH levels of acromegalics. The most likely interpretation of such finding is that circulating GH disrupts, for a negative feedback effect, hypothalamic opioid function and/or GH-releasing hormone (GHRH) producing neurons, through which opioids exert their action. To address this issue, we have evaluated in intact and hypophysectomized male rats the effect of a high-dose GH regimen on the hypothalamic stores of endogenous opioid peptides, beta-endorphin (beta-EP) and met-enkephalin (met-enk). Moreover we have evaluated in intact male rats the effect of exogenous GH on median eminence (ME) GHRH stores and the ability of FK 33-824 to stimulate GH and prolactin (PRL) secretion and of exogenous GHRH to induce GH secretion. Human GH (25 and 250 micrograms bid for 4 days) administered to hypophysectomized rats strikingly reduced beta-EP and met-enk-like immunoreactivity (LI) in the medial basal hypothalamus, the effect being already maximal with the lower hGH dose. The higher dose of hGH diminished, though to a lower extent, hypothalamic beta EP-LI content also in intact rats, and reduced GHRH-LI content in the ME. Despite these profound biochemical alterations, the GH responsiveness to GHRH and FK 33-824 administration was preserved, while the latter drug induced a lower PRL rise in GH-treated than in control rats.(ABSTRACT TRUNCATED AT 250 WORDS)