Ghrelin is an acylated peptide recently identified as the endogenous ligand for the growth hormone (GH) secretagogues receptor 1a (GHS-R1a) and is involved in a novel system for regulating GH release. To understand the long-term effects of ghrelin, here we constructed six myogenic expression vectors containing the cDNA of swine mature ghrelin (pGEM-wt-sGhln, pGEM-wt-hGhln), ghrelin mutant of $Ser^3$ with $Trp^3$ (pGEM-mt-sGhln, pGEM-mt-hGhln) and truncated ghrelin derivative (pGEM-tmtsGhln, pGEM-tmt-hGhln) encompassing the first 7 residues of ghrelin (including $Ser^3$ substituted with $Trp^3$) and adding a basic amino acid, Lys (K) in the C-terminus. The constructs, pGEM-wt-sGhln, pGEM-mt-sGhln and pGEM-tmt-sGhln were linked with the ghrelin leader sequence, while the pGEM-wt-hGhln, pGEM-mt-hGhln and pGEM-tmt-hGhln were linked with a leader sequence from the human growth hormone releasing hormone (hGHRH). Intramuscular injection of 200 ${\mu}g$ pGEM-wt-sGhln or pGEM-tmt-sGhln augmented growth over 3 weeks in normal rats and peaked at day 21 or 14 post-injection respectively, whose body weight gains were on average approximately 6% or 19% heavier over controls. However, other injectable vectors had no such enhanced growth effects. Our results suggested that the efficacy of the ghrelin leader sequence was more effective than that of hGHRH in our system. Moreover, the results indicated that skeletal muscle might have the ability to posttranslationally modify the in vivo expressed ghrelin. And the most strikingly, the short ghrelin analog seems to mimic the biological effects more efficiently when compared with the full-length ghrelin.
Purpose : The hope that arresting pubertal developement might increase final adult height has led to an attempt to use GnRH agonist (GnRHa) in children with early puberty and poor growth prognosis. We investigated the growth-promoting effect of GnRH agonists with or without growth hormone (GH) in girls with early puberty and decreased predicted adult height (PAH). Methods : Thirty five girls with advanced bone age and early pubertal signs were randomized for treatment for about 1 year with monthly GnRHa in group 1 (n=18), or with a combination of GH and GnRHa in group 2 (n=17). The following growth parameters were compared between groups, and the difference ($\Delta$) before and after treatment : chronological age (CA), bone age (BA), $\Delta$(BA-CA), height (HT), target height (TH), predicted adult height (PAH), $\Delta$ (TH-PAH), serum insulin-like growth factor (IGF-1) and insulin-like growth factor binding protein (IGFBP-3). Results : Before treatment, BA, TH, PAH Standard deviation scores (SDS), $\Delta$(TH-PAH) were not different between the two groups, but CA was higher in group 2 and $\Delta$(BA-CA) were higher in group 1 (P<0.05). After $1.06{\pm}0.93$ year of treatment, $\Delta$ (BA-CA) decreased and there were significant changes in PAH and $\Delta$ (TH-PAH), especially in group 2 (P<0.05 in group 1, and P<0.001 in group 2). In both groups, IGF-1 and IGFBP-3 were not different before and after treatment, but after treatment, IGF-1 level in group 2 was marginally higher than IGF-1 in group 1 (P<0.1). Conclusion : Compromised predicted adult height in girls with early puberty and advanced bone age was significantly improved with GnRH with/without GH treatment in the short-term period. The addition of GH to GnRHa results in a significant increase in PAH compared to GnRHa alone because GnRHa suppressed growth hormone-IGF-1 axis. For comparison of final adult height, further longitudinal follow-up will be needed.
Park, Seung-Joon;Park, Hee-Soon;Lee, Mi-Na;Sohn, Sook-Jin;Kim, Eun-Hee;Jung, Jee-Chang;Frohman, Lawrence A.;Kineman, Rhonda D.
The Korean Journal of Physiology and Pharmacology
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v.7
no.2
/
pp.79-84
/
2003
We have previously reported that expression of the somatostatin receptor subtypes, sst1-5, is differentially regulated by growth hormone (GH)-releasing hormone (GHRH) and forskolin (FSK), in vitro. GHRH binds to membrane receptors selectively located on pituitary somatotropes, activates adenylyl cyclase (AC) and increases sst1 and sst2 and decreases sst5 mRNA levels, without significantly altering the expression of sst3 and sst4. In contrast FSK directly activates AC in all pituitary cell types and increases sst1 and sst2 mRNA levels and decreases sst3, sst4 and sst5 expression. Two explanations could account for these differential effects: 1) GHRH inhibits sst3 and sst4 expression in somatotropes, but this inhibitory effect is masked by expression of these receptors in unresponsive pituitary cell types, and 2) FSK inhibits sst3 and sst4 expression levels in pituitary cell types other than somatotropes. To differentiate between these two possibilities, somatotropes were sequentially labeled with monkey anti-rat GH antiserum, biotinylated goat anti-human IgG, and streptavidin-PE and subsequently purified by fluorescent-activated cell sorting (FACS). The resultant cell population consisted of 95% somatotropes, as determined by GH immunohistochemistry using a primary GH antiserum different from that used for FACS sorting. Purified somatotropes were cultured for 3 days and treated for 4 h with vehicle, GHRH (10 nM) or FSK ($10{\mu}M$). Total RNA was isolated by column extraction and specific receptor mRNA levels were determined by semi-quantitative multiplex RT-PCR. Under basal conditions, the relative expression levels of the various somatostatin receptor subtypes were sst2>sst5>sst3=sst1> sst4. GHRH treatment increased sst1 and sst2 mRNA levels and decreased sst3, sst4 and sst5 mRNA levels in purified somatotropes, comparable to the effects of FSK on purified somatotropes and mixed pituitary cell cultures. Taken together, these results demonstrate that GHRH acutely modulates the expression of all somatostatin receptor subtypes within GH-producing cells and its actions are likely mediated by activation of AC.
Nou, V.;Inoue, H.;Lee, H.G.;Matsunaga, N.;Kuwayama, H.;Hidari, H.
Asian-Australasian Journal of Animal Sciences
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v.16
no.8
/
pp.1193-1198
/
2003
An increase in frequency of administration of exogenous growth hormone (GH) or GH-releasing hormone was reported to be a model to increase blood circulating insulin-like growth factor-1 (IGF-1) and to improve growth performance in animals. We have investigated the effect of twice daily administration of GH-releasing peptide-2 (GHRP-2) on growth performance, GH responsiveness and plasma insulin-like growth factor IGF-1 in swine. We administered to eight swine, 3 control and 5 treatment, a twice daily s.c. injections of GHRP-2 ($30{\mu}g/kg\;BW$) for a period of 10 days. Every day blood samples immediately taken before injections of GHRP-2 or saline, at 08:00 h and 16:00 h, were measured for IGF-1 concentrations. Blood samples for GH assay were collected every 20 min on days 1, 6 and 10, from 1 hour before and 3 h after GHRP-2 or saline injections at 08:00 h. GH peak concentrations and GH area under curve (GH AUC) on day 1, 6 and 10 in treatment group of swine were higher than those in control swine (p<0.05). Twice daily administration of GHRP-2 caused a significantly attenuation (p<0.05) of GH peak concentrations ($80.25{\pm}13.87$, $39.73{\pm}5.72$ and $27.57{\pm}6.06ng/ml$ for day 1, 6 and 10, respectively) and GH AUCs ($3,536.15{\pm}738.35$, $1,310.31{\pm}203.55$ and $934.37{\pm}208.99ng/ml$ for day 1, 6 and 10, respectively). However, there was no significant difference in GH peak concentration and GH AUC between day 6 and 10. Plasma IGF-1 concentration levels were higher in treatment than control group of swine (p<0.05) after 3 days of the treatment, and the levels reached a plateau from day 3 to 10 of experiment. Growth performance did not alter by GHRP-2 administration, even though a numerical increase of body weight gain and feed efficiency was observed. These results indicate that twice daily administration of GHRP-2 for 10 days in swine did not significantly influence on growth performance, caused an overall attenuation of GH response, and that elevation of plasma GH concentrations caused by GHRP-2 administration increased plasma IGF-1 concentrations, even though an attenuation of GH response was observed.
Biotechnology will play critical role in improving animal productivity. Animal growth rate and muscle deposition potential can be greatly improved by the application of biotechnology and biotechnological products. Administration of recombinant somatotropin (ST) or other compounds such as IGF-1 and growth hormone-releasing peptides (GHRPs) can enhance growth rate and carcass lean percentage. Gene transfer offers a powerful approach to manipulate endocrine system and metabolic pathways toward faster growth and better feed efficiency. Biotechnology is also extensively used for improving metabolism and activity of gut microorganisms for better nutrient digestibility. Knockout of growth-inhibiting genes such as myostatin results in considerable acceleration of body weight and muscle growth. Animal growth can also be improved by the use of gene therapy. Immunomodulation is another approach for efficient growth through controlling the activity of endogenous anabolic hormones. All the above aspects will be discussed in this review.
The purpose of this study was to determine the effect of energy supplement on responses of plasma insulin-like growth factor (IGF)-1 and IGF binding proteins (IGFBPs) to growth hormone-releasing peptide-2 (GHRP-2) administration in normal protein-fed wethers, and to observe the effect of GHRP-2 treatment on hepatic growth hormone (GH) receptor in well-fed wethers. Plasma IGF-1 and 39-42 kDa IGFBP-3 during the HENP (CP, crude protein 0.34 and TDN, total digestible nutrients 1.83 kg/day DM, dry matter intake) treatment period were higher than in the LENP (CP 0.32 kg and TDN 0.87 kg/day DM intake) period (P<0.05). The response of GH was stimulated by GHRP-2 ($12.5\;{\mu}g/kg$ body weight/day) administration during both of the feed treatment periods (P<0.05). The area under curve (AUC) increment and average concentration of GH (0-180 min) with GHRP-2 administration was higher during HENP treatment than LENP treatment (P<0.01). During the HENP treatment period from day 1 to day 7 of twice daily GHRP-2 treatment, the plasma IGF-1 increment was increased on days 2, 6 and 7 of GHRP-2 administration (P<0.05). On the basis of ligand blotting, the proportions of plasma 39-43 kDa IGFBP-3 during the HENP treatment period only showed a significant difference on days 6 and 7 with GHRP-2 administration. No significant difference in the specific binding of $^{125}I-labeled$ oGH to hepatic membranes was detected between the saline and GHRP-2 treatments of the HENP-fed wethers. These results suggest that the nutritional balance between energy and protein may affect the endogenous GH / IGF-1 axis as well as plasma IGFBP-3 levels.
The factors involved in the initial neoplastic transformation and subsquent growth of uterine fibroid are poorly understood. The reduction in uterine fibroid volume associated with the chronic administration of the mechanisms mediating the decrease in fibroid volume in GnRH-a treated patients are poorly defined. The purpose of this study was to determine the proliferating cell nuclear antigen(PCNA) in fibroid from-women pretreated with GnRH analogue(GnRH-a) compared with controls. Tissue was obtained from 16 premenopausal women with uterine fibroid who received GnRH-a(D-Trp6-GnRH) intramusculary every 28 days for four injections. The mean proliferating index(PI) in patients with uterine fibroids was $2.25{\pm}0.9$, and in controls was $8.82{\pm}1.8$(P<0.001). The proliferating index was not corrleated with the reduction of fibroid volume. In this clinical study, although hypoestrogenism may be the main factor that reduce the volume of fibroid, other factors are also considered to be involved in that process. And the regrowth of uterine fibroid may be affected by increased production of PCNA after stopping GnRH-a.
Kim, Doosoo;Cho, Sung-Yoon;Maeng, Se-Hyun;Yi, Eun Sang;Jung, Yu Jin;Park, Sung Won;Sohn, Young Bae;Jin, Dong-Kyu
Clinical and Experimental Pediatrics
/
v.55
no.12
/
pp.481-486
/
2012
Purpose: Precocious puberty is defined as breast development before the age of 8 years in girls. The present study aimed to reveal the diagnosis of Korean girls referred for precocious puberty and to compare the constitutional and endocrinological features among diagnosis groups. Methods: The present study used a retrospective chart review of 988 Korean girls who had visited a pediatric endocrinology clinic from 2006 to 2010 for the evaluation of precocious puberty. Study groups comprised fast puberty, true precocious puberty (PP), pseudo PP, premature thelarche, and control. We determined the height standard deviation score (HSDS), weight standard deviation score (WSDS), and body mass index standard deviation score (BMISDS) of each group using the published 2007 Korean growth charts. Hormone tests were performed at our outpatient clinic. Results: The PP groups comprised fast puberty (67%), premature thelarche (17%), true PP (15%), and pseudo PP (1%). Advanced bone age and levels of estradiol, basal luteinizing hormone (LH), and peak LH after gonadotropin-releasing hormone stimulation testing were significantly high in the fast puberty and true PP groups compared with the control group. HSDS, WSDS, and BMISDS were significantly higher in the true PP group than in the control group (P<0.05). Conclusion: The frequent causes of PP were found to be fast puberty, true PP, and premature thelarche. Furthermore, BMISDS were significantly elevated in the true PP group. Therefore, we emphasize the need for regular follow-up of girls who are heavier or taller than others in the same age group.
Cho, Eun Seok;Park, Da Hye;Kim, Byeong-Woo;Jung, Won Youg;Kwon, Eun Jung;Kim, Chul Wook
Asian-Australasian Journal of Animal Sciences
/
v.22
no.3
/
pp.307-312
/
2009
The study of candidate genes is an important tool to identify genes associated with economic traits. Three genes were selected to study the association between polymorphism and economic traits and breed of pigs. Growth hormone releasing hormone (GHRH) takes part in growth metabolism and is one of the candidate genes known to be highly associated with economic traits in pigs. Heart fatty acid-binding protein (H-FABP) is related to back fat thickness and intramuscular fat (IMF) content, and myogenin (MYOG) is associated with the amount of growth rate and lean yield in pigs. By PCR-RFLP analysis, the association between the genotypes of the three genes and the average daily gain, back fat thickness, feed conversion, body length and meat percent in 352 pigs (112 Duroc pigs, 132 Landrace pigs and 108 Yorkshire pigs) were analyzed. GHRH polymorphisms showed differences depending on breed (p<0.01) and were associated with meat percent. H-FABP polymorphisms also showed significant differences among breeds and sex (p<0.01), and were highly associated with average daily gain, feed conversion and back fat thickness (p<0.01) and even showed an association with meat percent (p<0.05). However, the MYOG gene showed no significant effect in this study. These results reconfirmed that GHRH and H-FABP are potential major genes or markers for economic traits.
Park, Seung-Joon;Yang, In-Myung;Yim, Sung-Vin;Chung, Joo-Ho;Jung, Jee-Chang;Ko, Kye-Chang;Kim, Young-Seol;Choi, Young-Kil
The Korean Journal of Physiology and Pharmacology
/
v.2
no.1
/
pp.101-108
/
1998
We investigated the effect of ${\alpha}-subunit$ of the stimulatory GTP-binding protein ($G{\alpha}_s$) gene mutation on the expression of the thyrotropin-releasing hormone (TRH) receptor (TRH-R) gene in GH3 cells and in growth hormone (GH)-secreting adenomas of acromegalic patients. In the presence of cyclohexicmide, forskolin and isobutylmethylxanthine, cholera toxin, and GH-releasing hormone (GHRH) decreased rat TRH-R (rTRH-R) gene expression by about 39%, 43.7%, and 46.7%, respectively. Transient expression of a vector expressing mutant-type $G{\alpha}_s$ decreased the rTRH-R gene expression by about 50% at 24 h of transfection, whereas a wild-type $G{\alpha}_s$ expression vector did not. The transcript of human TRH-R (hTRH-R) gene was detected in 6 of 8 (75%) tumors. Three of them (50%) showed the paradoxical GH response to TRH and the other three patients did not show the response. The relative expression of hTRH-R mRNA in the tumors from patients with the paradoxical response of GH to TRH did not differ from that in the tumors from patients without the paradoxical response. Direct PCR sequencing of $G{\alpha}_s$ gene disclosed a mutant allele and a normal allele only at codon 201 in 4 of 8 tumors. The paradoxical response to TRH was observed in 2 of 4 patients without the mutation, and 2 of 4 patients with the mutation. The hTRH-R gene expression of pituitaty adenomsa did not differ between the tumors without the mutation and those with mutation. The present study suggests that the expression of TRH-R gene is not likely to be a main determinant for the paradoxical response of GH to TRH, and that $G{\alpha}_s$ mutation may suppress the gene expression of TRH-R in GH-secreting adenoma. However, a certain predisposing factor(s) may play an important role in determining the expression of TRH-R.
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