Somatostatin inhibition of teleost growth hormone secretion.

Pituitary glands of a teleost, tilapia (Sarotherodon mossambicus), were incubated in vitro in the presence or absence of synthetic somatostatin. After a 4-hr incubation period, the growth hormone (GH) concentration of the incubation medium was determined using a highly specific homologous radioreceptor assay. The presence of somatostatin in the incubation medium at concentrations of 0.1 to 2.0 μg/ml produced a dose-dependent inhibition of the spontaneous release of GH. These results suggest that GH secretion in teleosts may be influenced by a somatostatin-like peptide which acts to suppress the release of GH from the pituitary.     

A sexually dimorphic pattern of growth hormone secretion in the elderly.

In rodents, the sexually dimorphic pattern of pulsatile GH secretion is an important determinant of growth, liver enzyme function and insulin-like growth factor I (IGF-I) expression. Whether this difference is present in humans at different ages is unclear. We studied GH secretory patterns in the elderly by constructing 24-h serum GH profiles in 45 male and 38 female (age, 59.4-73.0 yr) volunteers and related patterns to IGF-I, IGF-binding protein-3 (IGFBP-3), and GH-binding protein levels; body mass index; and waist/hip ratio. Serum GH concentrations were measured in samples drawn at 20-min intervals and analyzed using a sensitive chemiluminescent assay (Nichols Institute Diagnostics: sensitivity, 0.036 mU/L). The 24-h serum GH profiles were analyzed using a concentration distribution method to determine GH peak and trough levels, spectral analysis, and assessment of serial irregularity by approximate entropy (ApEn). There was a highly significant difference in mean 24-h serum GH concentrations in females compared to males (males, 0.88 mU/L; females, 1.31 mU/L; P = 0.009) as a result of significantly higher trough GH levels (males, 0.04 mU/L; females, 0.16 mU/L; P < 0.001). Peak values were not significantly different. Serum IGF-I levels were significantly higher in males (males, 162.4 ng/mL; females, 87.8 ng/ mL; P < 0.001). Peak GH values were related to serum IGF-I levels (males: r = 0.39; P = 0.009; females: r = 0.5; P = 0.002), whereas trough GH levels were not. IGFBP-3 levels were similar and related to GH peaks only in males (r = 0.32; P = 0.03). GH was secreted with a dominant periodicity of 200 min in males and 280 min in females (P < 0.025). The proportion of time taken up by regular oscillatory activity was less in females (females, 11.1%; males, 14.7%; P = 0.01). GH secretion assessed by ApEn was more disordered in females (males, 0.60; females, 0.81; P < 0.001), and increasing disorder was associated with lower IGF-I levels. Body mass index was negatively related to GH in both sexes. In males, trough values were the major determinant (r = -0.31; P = 0.04), whereas in females, the peak value was the major determinant (r = 0.35; P = 0.04). Trough GH levels were inversely related in both sexes to waist/hip ratio (males: r = -0.40; P = 0.006; females: r = -0.44; P = 0.006) and to increasing secretory disorder (ApEn; r = -0.46; P < 0.001). These data demonstrate a sexually dimorphic pattern of GH secretion in the elderly.     

Evidence that the regulation of growth hormone secretion is mediated predominantly by a growth hormone releasing factor

Spontaneous pulsatile growth hormone (GH) secretion and stress-induced suppression of GH was examined in chronically cannulated male rats with electrolytic lesions of the periventricular preoptic anterior hypothalamic area (PO/AHA) where somatostatin neurons innervating the median eminence are known to be located. Median eminence somatostatin was depleted in lesioned animals by 85%. Bursts of GH secretion occurred more frequently than in sham-lesioned animals (1.9 +/- 0.2 vs. 2.6 +/- 0.2 h, respectively, p less than 0.025), however, average concentrations of GH were reduced by lesions (118.4 +/- 11.6 vs 192.3 +/- 28.4 ng/ml, p less than 0.025). Suppression of GH by stress was unaffected by PO/AHA lesions. It is concluded that somatostatin plays only minor roles in the generation of GH troughs during rhythmic GH secretion, and in the suppression of GH during stress. Inhibition of GH releasing factor secretion, therefore, is presumed to be the likely method by which GH suppression is achieved in these physiological situations.     

Pituitary secretion of growth hormone in response to opioid peptides and opiates is mediated through growth hormone-releasing factor

Passive immunization of rats with an antiserum raised against rat growth hormone-releasing factor (GRF) completely inhibited the growth hormone (GH) response to morphine and beta-endorphin but did not alter the prolactin (PRL) response to those two stimuli. These results demonstrate that opiate and opioid peptide stimulation of pituitary GH secretion is mediated through hypothalamic GRF and presents an animal model in which the stimulated secretion of GH and PRL can be specifically dissociated.     

Galanin secretion from anterior pituitary cells in vitro is regulated by dopamine, somatostatin, and thyrotropin-releasing hormone.

Lactotrophs, somatotrophs, and thyrotrophs have been shown to contain immunoreactive galanin. Furthermore, estrogen stimulates galanin mRNA and peptide levels in the rat anterior pituitary, particularly within lactotrophs. To determine whether galanin is released from the anterior pituitary in a regulated manner, we used cultured pituitary cells from male and ovariectomized Fischer 344 rats implanted with estrogen-containing capsules. Anterior pituitary cells (5 x 10(5) cells/well) were challenged (0.5-3 h) with hypothalamic factors known to regulate anterior pituitary hormone secretion, and medium galanin levels were measured by RIA. In female pituitary cells, galanin secretion was inhibited by dopamine (10 and 100 nM) and stimulated by TRH (20 and 100 nM). Although galanin release was significantly lower in male pituitary cells, dopamine and TRH inhibited and stimulated galanin secretion, respectively. Medium galanin levels were also significantly reduced by somatostatin (5 nM) in both female and male cells. The pattern of PRL release in response to dopamine, TRH, and somatostatin was similar to that observed for galanin, regardless of the sex of the pituitary donor. Although galanin has been localized in somatotrophs, 5 nM GH-releasing hormone (GRF) failed to alter galanin release in male as well as female pituitary cells; GH secretion was significantly increased by GRF. LHRH (5 nM) and CRF (5 nM) failed to alter galanin release in vitro. We conclude that in estrogen-exposed pituitary cells obtained from male and ovariectomized Fischer 344 rats: 1) galanin secretion is inhibited by dopamine and somatostatin, and stimulated by TRH; 2) GRF, LHRH, and CRF do not regulate galanin release in these cells; and 3) the profile of the regulated pathway for galanin release suggests that the primary location of galanin is the lactotroph, probably within secretory granules.     

Pituitary growth hormone network responses are sexually dimorphic and regulated by gonadal steroids in adulthood

There are well-recognized sex differences in many pituitary endocrine axes, usually thought to be generated by gonadal steroid imprinting of the neuroendocrine hypothalamus. However, the recognition that growth hormone (GH) cells are arranged in functionally organized networks raises the possibility that the responses of the network are different in males and females. We studied this by directly monitoring the calcium responses to an identical GH-releasing hormone (GHRH) stimulus in populations of individual GH cells in slices taken from male and female murine GH-eGFP pituitary glands. We found that the GH cell network responses are sexually dimorphic, with a higher proportion of responding cells in males than in females, correlated with greater GH release from male slices. Repetitive waves of calcium spiking activity were triggered by GHRH in some males, but were never observed in females. This was not due to a permanent difference in the network architecture between male and female mice; rather, the sex difference in the proportions of GH cells responding to GHRH were switched by postpubertal gonadectomy and reversed with hormone replacements, suggesting that the network responses are dynamically regulated in adulthood by gonadal steroids. Thus, the pituitary gland contributes to the sexually dimorphic patterns of GH secretion that play an important role in differences in growth and metabolism between the sexes.     

Grtp1, a novel gene regulated by growth hormone

An in vitro model of GH-responsive cells was subjected to microarray analysis to identify a novel gene regulated by GH. This 258 amino acid protein, we term GH Regulated TBC Protein-1 (GRTP1), contains the TBC signature motif of GTPase activator proteins of Rab-like small GTPases. Northern blot analysis revealed a 1.3 kb major mRNA species, most abundant in testes. TaqMan assay confirmed that in the mouse, Grtp1 is expressed at highest levels in testes, with lesser abundance in intestine, kidney, lung, and liver. In the testis, expression of Grtp1 significantly increases post-pubertally. Administration of GH to mice increased levels of GRTP1 mRNA in testes (140%), but decreased GRTP1 mRNA abundance in kidney (50%) and liver (25%). Grtp1 was localized to mouse proximal chromosome 8. Orthologs of this protein are present in human, mouse, rat, and drosophila suggesting that GRTP1 has an important biological role(s).     

Greater conservation of somatolactin, a presumed pituitary hormone of the growth hormone/prolactin family, than of growth hormone in teleost fish

From pituitary cDNA libraries of Atlantic cod and chum salmon, cDNA clones coding for somatolactin (SL), a presumed pituitary hormone belonging to the growth hormone (GH)/prolactin (PRL) family, were isolated and characterized. The 1.3-kb cod SL mRNA was composed of a greater than 0.25-kb 5' untranslated region, a coding region for the precursor of 235 amino acids (aa), a 0.14-kb 3' untranslated region, and a poly(A) tail. The 2.5-kb salmon SL mRNA had a less than 0.1-kb 5' untranslated region, a precursor (233 aa) coding region, a 1.6-kb 3' untranslated region, and a poly(A) tail. A signal peptide of 26 and 24 aa was found in the SL precursor of cod and salmon, respectively. Thus, the mature SLs of these fish are composed of 209 aa. Two potential N-glycosylation sites were identified in cod SL, whereas no site could be found in the salmon. A comparison of amino acid sequences of the three SLs so far isolated indicated six Cys residues to be in homologous positions to those in GH and PRL, and one Cys residue to be characteristically present in SL. Among cod, salmon, and flounder, greater colinearity of amino acid sequences was noted in SLs than in GHs. The identities of the SL amino acid sequences were between 73 and 81% as compared to 58-62% for the corresponding GHs, indicating greater conservation of SL than GH.     

Opiate modulation of growth hormone secretion is compromised during the steroid-induced luteinizing hormone surge.

Earlier studies from our laboratory have shown that treatments with gonadal steroids which cause a surge of luteinizing hormone (LH) blunt the effects of morphine on LH secretion, locomotor activity, nociception and temperature regulation. The present study was conducted to determine if the growth hormone (GH) response to morphine sulfate (MS) was also affected during steroid-induced LH surges. Adult ovariectomized female rats were primed with estradiol benzoate (EB: -49 h prior to P4 or oil injection) and/or progesterone (P4; 10.00 h). Seven and one half hours after P4 treatment, at the time of the steroid-induced LH surge, the GH response to an intravenous dose of 0.5, 2.0 or 5.0 mg/kg MS was determined. A GH peak response occurred at 15 min after drug administration and was maximal at the dose of 2 mg/kg MS in all groups. Although the timing of the GH rise was not altered in either EB/oil- or EB/P4-treated animals, a significant blunting of MS-induced GH secretion was observed across all doses of MS with a greater reduction being observed in EB/P4-treated animals. In a second study the GH response to opiates was investigated prior to and following the steroid-induced LH surge to determine if the suppression of opiate-induced GH secretion was confined to the time of the preovulatory LH surge. Before the LH surge (2.5 h), a mild suppression of opiate-induced GH secretion was observed only in EB/oil-treated animals. After the LH surge (13 h after P4 administration), morphine-induced GH secretion was similar in all groups.(ABSTRACT TRUNCATED AT 250 WORDS)     

Growth regulation in the gobiid teleost, Gillichthys mirabilis: roles of growth hormone, hepatic growth hormone receptors and insulin-like growth factor-I.

Studies of the teleost Gillichthys mirabilis were undertaken to assess the role of GH in regulating hepatic GH receptors, insulin-like growth factor-I (IGF-I) activity and cartilage growth. Hypophysectomized G. mirabilis were injected with saline vehicle or with 10, 100 or 1000 ng tilapia GH (tGH)/g every other day for 2-3 weeks. Growth, judged by increased body weight and length, was inhibited by hypophysectomy and stimulated by increasing tGH doses. Hepatic GH receptors, measured by 125I-labelled tGH binding, were decreased 50% by hypophysectomy. An additional dose-dependent reduction in binding was observed 24 h after tGH injection, but MgCl2 stripping of membranes suggested receptor occupation by exogenous tGH. IGF-I activity, measured by 35SO4 incorporation into oral cartilage explants in vitro, was decreased 50% by hypophysectomy and increased to 200% of intact control levels after injection of 1000 ng tGH/g. In a second experiment, 35SO4 incorporation by oral cartilage from hypophysectomized fish injected with 10, 100 or 1000 ng tGH/g was stimulated to intact control levels. Effects of feeding and tGH injection on in-vitro responsiveness of oral cartilage to recombinant bovine IGF-I (rbIGF-I; 10-1000 ng/ml) were also assessed. Oral cartilage from fed fish showed a parabolic dose-response curve, whereas oral cartilage from starved fish had a lower basal rate of 35SO4 incorporation and a linear dose-response relationship. Oral cartilage from hypophysectomized G. mirabilis showed a significantly attenuated response to rbIGF-I which was restored by tGH injection, suggesting that the GH status of the animal is important for sensitivity of target tissue to IGF-I. Because of its similarity to other vertebrate systems, G. mirabilis presents a good teleost model of growth regulation and of the functions and interactions of GH and IGF-I.     

Galanin is a physiological regulator of spontaneous pulsatile secretion of growth hormone in the male rat.

To determine whether galanin (GAL), a 29-amino acid neuropeptide, plays a role in the physiological regulation of the pulsatile secretion of GH and PRL in the male rat, secretory patterns of both hormones were studied in freely moving animals after GAL passive immunoneutralization. Adult male Sprague-Dawley rats were equipped with iv and intracerebroventricular catheters. After 7 days, 3 microliters of a specific GAL antiserum (GAL-AS) or normal rabbit serum (NRS; controls) were infused in the third ventricle of 10 rats, 25 and 1 h before the animals were bled every 15 min for 6 h (1000-1600 h). Plasma GH and PRL concentrations were measured by RIA, and the hormonal secretory patterns were analyzed by the PULSAR program. Control rats, treated with NRS, displayed typical GH secretion, with pulses of high amplitude (167 +/- 27 ng/ml) and low frequency (2.4 +/- 0.2 pulses/6 h), separated by periods of low trough levels (3.8 +/- 0.6 ng/ml). Rats treated with GAL-AS had altered pulsatile GH secretion. Pulse height was markedly reduced (77 +/- 15 ng/ml; P less than 0.01 vs. controls), and peak frequency was higher (3.6 +/- 0.5 pulses/6 h; P less than 0.05), while GH baseline levels and integrated GH secretion over the 6-h sampling period remained unaltered. Injection of rat GH-releasing hormone (1 microgram/rat, iv) caused a similar GH stimulation in both groups of rats, as determined by the peak GH response at 5 min (368 +/- 112 vs. 342 +/- 81 ng/ml) or by the integrated GH response over 1 h (5.13 +/- 1.30 vs. 4.77 +/- 1.15 micrograms.min/ml in NRS- and GAL-AS-treated rats, respectively; P less than 0.05). In contrast to GH, pulsatile secretion of PRL was not affected by the GAL-AS treatment. These results indicate that GAL is a physiological regulator of spontaneous pulsatile secretion of GH, but not PRL, in the male rat. The influence of GAL on GH secretion appears to be exerted within the hypothalamus, mainly by a stimulation of GRF secretion. However, the changes in GH pulse frequency observed after GAL immunoneutralization suggest that GAL might also influence the somatostatin inhibitory tone.     

Cachexia in rheumatoid arthritis is not explained by decreased growth hormone secretion

OBJECTIVE: Patients with rheumatoid arthritis (RA) lose body cell mass (BCM) by unknown mechanisms. Since the loss of BCM in normal aging individuals parallels the characteristic age-related decline in growth hormone (GH) secretion, this study was carried out to determine whether further decreased GH secretion plays a role in the pathogenesis of this loss of BCM in RA patients, termed "rheumatoid cachexia." METHODS: GH secretory kinetics were determined by deconvolution analysis in 16 patients with RA and 17 healthy controls matched for age (mean +/- SD 45.4 +/- 13.2 years and 47.1 +/- 14.6 years, respectively), sex, race, and body mass index. Blood samples were obtained every 20 minutes for 24 hours. Body composition was ascertained using total-body potassium (TBK) as a measure of BCM and dual x-ray absorptiometry to determine fat mass. RESULTS: BCM was reduced in patients with RA compared with healthy controls (mean +/- SD gm TBK 79.5 +/- 9.5 versus 94.9 +/- 11.9; P < 0.0005), but there was no difference in fat mass. GH kinetic parameters in patients with RA did not differ from those in controls. CONCLUSION: These findings suggest that GH kinetics are unaltered in RA patients compared with healthy subjects; thus, GH deficiency does not account for rheumatoid cachexia.     

Stimulation of prolactin and growth hormone secretion by muscimol, a gamma-aminobutyric acid agonist.

The alteration in circulating levels of PRL, GH, TSH, and cortisol was studied after the oral administration of muscimol (3-hydroxy-5-aminomethylisoxazole) to human subjects with Huntington's disease (n = 4) and chronic schizophrenia (n = 5). PRL levels rose significantly in a dose-dependent fashion within a 120-min time interval. GH rose significantly but modestly over the same time interval, whereas TSH and cortisol levels remained unchanged. Since muscimol is thought to be a potent and specific gamma-aminobutyric acid agonist, these data indicate that gamma-aminobutyric acid-mediated neural transmission may function to stimulate the release of plasma PRL and GH in human subjects.     

Ghrelin secretion in humans is sexually dimorphic,suppressed by somatostatin,and not affected by the ambient growth hormone levels

We studied plasma ghrelin and GH concentrations over a 24-h period in young healthy men and women and in patients with acromegaly. Healthy subjects were restudied after administration of GH-lowering agents, octreotide or GHRH antagonist. Ghrelin concentrations in women studied during the late follicular stage of the cycle were about 3-fold higher than in men. Suppression of GH secretion by GHRH antagonist did not alter ghrelin concentration profiles. In the presence of high GH levels (acromegaly), ghrelin levels were similar to those found in healthy men. Administration of somatostatin analog octreotide suppressed both GH and ghrelin concentration profiles. We conclude that: 1) ghrelin secretion is sexually dimorphic in humans, with women in the late follicular stage having higher levels than men; 2) ghrelin secretion is suppressed by somatostatin; and 3) GH has no influence over ghrelin secretion.     

Inhibition of the acetylcholine-induced secretion of bovine growth hormone by somatostatin.

The effects of calcium deprivation, somatostatin and verapamil on the stimulation of growth-hormone release and the alteration of pituitary metabolism in response to acetylcholine were investigated. Calcium deprivation decreased the rises in growth-hormone secretion and in cyclic GMP content in response to 25 μM acetylcholine but did not prevent the increased incorporation of ³²P into phosphatidyl inositol. Somatostatin (1 μg/ml) prevented the rise in growth-hormone secretion and inhibited the efflux of ⁴⁵Ca in response to acetylcholine (25 μM) but did not modify the increase in cyclic GMP content or phosphatidyl inositol labelling. Verapamil (50 μM) did not affect any of the responses to acetylcholine (25 μM). The data suggest that acetylcholine stimulates calcium mobilization from tissue stores and that somatostatin can prevent this mobilization. The relevance of this to the inhibition of secretion by somatostatin is discussed.     

Physiological control of growth hormone secretion by thyrotrophin-releasing hormone in the domestic fowl

Immature cockerels (4- to 5-weeks old) were passively immunized, with antiserum raised in sheep, against thyrotrophin-releasing hormone (TRH). The administration of TRH antiserum (anti-TRH) at doses of 0.5, 1.0 or 2.0 ml/kg lowered, within 1 h, the basal concentration of plasma GH for at least 24 h. The administration of normal sheep serum had no significant effect on the GH concentration in control birds. Although the GH response to TRH (1.0 or 10.0 micrograms/kg) was not impaired in birds treated 1 h previously with anti-TRH, prior incubation (at 39 degrees C for 1 h) of TRH (20 micrograms/ml) with an equal volume of anti-TRH completely suppressed the stimulatory effect of TRH (10 micrograms/kg) on GH secretion in vivo. These results suggest that TRH is physiologically involved in the hypothalamic control of GH secretion in the domestic fowl.