Growth Hormone mRNA Levels Research Articles

Is really endogenous ghrelin a hunger signal in chickens? Association of GHSR SNPs with increase appetite, growth traits, expression and serum level of GHRL, and GH

Chicken growth hormone secretagogue receptor (GHSR) is a receptor for ghrelin (GHRL), a peptide hormone produced by chicken proventriculus, which stimulates growth hormone (GH) release and food intake. The purpose of this study was to search for single nucleotide polymorphisms (SNPs) in exon 2 of GHSR gene and to analyze their effect on the appetite, growth traits and expression levels of GHSR, GHRL, and GH genes as well as serum levels of GH and GHRL in Mandara chicken. Two adjacent SNPs, A239G and G244A, were detected in exon 2 of GHSR gene. G244A SNP was non-synonymous mutation and led to replacement of lysine amino acid (aa) by arginine aa, while A239G SNP was synonymous mutation. The combined genotypes of A239G and G244A SNPs produced three haplotypes; GG/GG, GG/AG, AG/AG, which associated significantly (P<0.05) with growth traits (body weight, average daily gain, shank length, keel length, chest circumference) at age from >4 to 16w. Chickens with the homozygous GG/GG haplotype showed higher growth performance than other chickens. The two SNPs were also correlated with mRNA levels of GHSR and GH (in pituitary gland), and GHRL (in proventriculus and hypothalamus) as well as with serum level of GH and GHRL. Also, chickens with GG/GG haplotype showed higher mRNA and serum levels. This is the first study to demonstrate that SNPs in GHSR can increase appetite, growth traits, expression and level of GHRL, suggesting a hunger signal role for endogenous GHRL.

Different effects of growth hormone and fasting on the induction patterns of two hormone-sensitive lipase genes in red seabream Pagrus major

Growth hormone (GH) increases phosphorylation and mRNA levels of hormone-sensitive lipase (HSL) in the livers of some marine teleosts. The hepatic GH-HSL axis appears to play important roles in fasting-induced lipolysis. However, it is not known whether GH exerts similar effects on HSL in fish adipose tissues. Functional differentiation of two fish-specific HSL isoforms (HSL1 and HSL2) also remains unclear. The present study seeks to address two unanswered questions about fish lipolysis using red seabream (Pagrus major): (1) Does GH increase phosphorylation and mRNA levels of HSL in adipose tissue? (2) How do GH and fasting affect mRNA levels of two HSL isoform genes in the liver and adipose tissue? To this end, we first cloned HSL1 and HSL2 cDNAs and investigated their tissue distribution. Transcripts of both HSLs and HSL1 proteins were abundant in the visceral adipose tissue, gonads, and liver, suggesting the important role of HSL in adipose tissue lipolysis. HSL2 transcript levels were 20–65% those of HSL1 except in the skin, and HSL2 proteins were not detected by our in-house antisera. Ex vivo administration of GH increased HSL1 phosphorylation, non-esterified fatty acid (NEFA) release, and levels of HSL1 and HSL2 mRNA in both the liver and visceral adipose tissue. Hepatic HSL2 mRNA was particularly sensitive to GH administration and sometimes exceeded HSL1 mRNA levels with up to 13-fold induction. In contrast, fasting for 4 and 7d increased HSL1 mRNA levels, but had only marginal effects on HSL2 mRNA levels in both adipose tissue or liver. We concluded that GH would increase HSL mRNAs during adipose tissue lipolysis in red seabream; however, GH and fasting result in different induction ratio of two HSL isoform genes, suggesting that other hormone(s) also contributes to fasting-induced lipolysis.

Feeding rates affect growth, intestinal digestive and absorptive capabilities and endocrine functions of juvenile blunt snout bream Megalobrama amblycephala.

This study aimed to investigate the optimal feeding rate for juvenile blunt snout bream (average initial weight 23.74 ± 0.09 g) based on the results on growth performance, intestinal digestive and absorptive capabilities and endocrine functions. A total of 840 fish were randomly distributed into 24 cages and fed a commercial feed at six feeding rates ranging from 2.0 to 7.0% body weight (BW)/day. The results indicated that weight gain rate increased significantly (P < 0.05) as feeding rates increased from 2.0 to 5.0% BW/day, but decreased with the further increasing feeding rates (P > 0.05). Protein efficiency ratio and nitrogen and energy retention all showed a similar trend. However, feed conversion ratio increased significantly (P < 0.05) with increasing feeding rates. Feeding rates have little effects (P > 0.05) on whole-body moisture, ash and protein contents, but significantly (P < 0.05) affect both lipid and energy contents with the highest values both observed in fish fed 4.0% BW/day. In addition, moderate ration sizes (2.0-4.0% BW/day) resulted in the enhanced activities of intestinal enzymes, including lipase, protease, Na(+), K(+)-ATPase, alkaline phosphatase and creatine kinase. Furthermore, the mRNA levels of growth hormone, insulin-like growth factors-I, growth hormone receptor and neuropeptide all increased significantly (P < 0.05) as feeding rates increased from 2.0 to 5.0% and 6.0% BW/day, but decreased significantly (P < 0.05) with the further increase in feeding rates, whereas both leptin and cholecystokinin expressions showed an opposite trend. Based on the broken-line regression analysis of SGR against feeding rates, the optimal feeding rate for juvenile blunt snout bream was estimated to be 4.57% BW/day.

Insulin-like growth factor 1 (IGF-1) regulates prolactin, growth hormone, and IGF-1 receptor expression in the pituitary gland of the gilthead sea bream Sparus aurata.

The role of insulin-like growth factor 1 (IGF-1) on regulation of growth hormone (GH) and prolactin (PRL) as well as the possible involvement of IGF-1 receptor subtype a (IGF-1Ra) mRNA was assessed in juvenile specimens of Sparus aurata. IGF-1Ra was successfully cloned, and active receptor domains were localized in its mRNA precursor. Also, phylogenetic analysis of the protein sequence indicated a closer proximity to IGF-1Ra isoform found in zebrafish and other teleosts, than to the isoform IGF-1Rb. The most abundant presence of IGF-1Ra mRNA was detected in white muscle, whereas head kidney showed the lowest gene expression among 24 different studied tissues. Pituitaries of juvenile specimens of S. aurata were incubated in vitro with different doses of IGF-1 (0, 1, 100, and 1000 ng mL(-1)) during a period of 10 h. Total RNA with a high quality could be obtained from these pituitaries. PRL mRNA expression significantly increased with increasing IGF-1 doses. Similarly, IGF-1Ra mRNA increased its expression in response to IGF-1. However, GH mRNA levels decreased in a dose-dependent manner after IGF-1 treatment. The contradictory responses of GH and PRL expressions to IGF-1 in our experiment are possibly mediated by IGF-1Ra presence on the somatotrophs and prolactotrophs. The increase in IGF-1Ra mRNA levels may be related to the proper activation of the PI3-K/Akt signal transduction pathways which are normally involved in GH and PRL regulation.

Transcription of growth hormone mRNA as a molecular marker of egg quality in Siberian sturgeon (Acipenser baerii)

The Siberian sturgeon (Acipenser baerii) is an economically important species. Sturgeon stockassessment during fingerling production is considered to be one of the most difficult phases ofhatchery rearing. The present study investigated the expression of growth hormone (GH)mRNA in unfertilized eggs, fertilized eggs, eyed eggs (2 day before hatching), non-viable eggsand newly-hatched Siberian sturgeon. Knowledge about the expression of GH during egg andembryo development can help determine the quality of the eggs. This can help predict larvalviability and, to some extent, directly determine the growth and survival of young fish. Little isknown about the effect of this hormone on egg quality in sturgeon. The present study analyzedrelative GH mRNA expression using the 2-ΔΔCT method. Ribosomal protein L6 (RPL6)transcripts were used as the housekeeping gene for normalization of GH mRNA transcription.The highest levels of GH mRNA were found in eyed eggs and the lowest levels were detected innon-viable eggs (P< 0.05). These findings suggest GH mRNA as a potential marker for eggquality in Siberian sturgeon.

Growth enhancement of rainbow trout (Oncorhynchus mykiss) by passive immunization against somatostatin-14

Juvenile rainbow trout (Oncorhynchus mykiss) were passively immunized by intraperitoneal immunization against somatostatin-14 (SS-14) using an antibody originating from egg-laying chicken (Gallus domesticus). Fish were immunized weekly (0, 7, 14, 21, 28, 35 days) with chicken egg yolk-derived immunoglobulin (IgY) against SS-14 (1:25 IgY, 5 mg mL−1), and growth performance, feed utilization as well as plasma concentrations and mRNA levels of growth hormone (GH) and insulin-like growth factor I (IGF-I) were compared to the control group that received placebo immunization with PBS. Passive immunization significantly increased weight gain of treated fish (67.7 ± 7.4 g) compared to the control group (40.1 ± 2.0 g) after 35 days (p 0.05). In both groups, GH levels decreased over the 35 days of the experiment (p 0.05). There was no indication of a cumulative, long-lasting effect of repeated immunization on GH or IGF-I plasma concentrations or mRNA expression. The present study shows that a passive immunization of rainbow trout against SS-14 using a chicken egg yolk-derived SS-14 antibody could increase growth rate and improved FCR.

Acclimation to different environmental salinities induces molecular endocrine changes in the GH/IGF-I axis of juvenile gilthead sea bream (Sparus aurata L.).

To assess the role of the GH/IGF-I axis in osmotic acclimation of the gilthead seabream Sparus aurata, juvenile specimens were acclimated to four environmental salinities: hyposmotic (5 ‰), isosmotic (12 ‰) and hyperosmotic (40 and 55 ‰). The full-length cDNAs for both pituitary adenylate cyclase-activating peptide (PACAP) and prepro-somatostatin-I (PSS-I), the precursor for mature somatostatin-I (SS-I), were cloned. Hypothalamic PACAP and PSS-I, hypophyseal growth hormone (GH) and prolactin (PRL), and hepatic insulin-like growth factor-I (IGF-I) mRNA expression levels were analyzed in the four rearing salinities tested. PACAP and IGF-I mRNA values increased significantly in response to both 5 and 55 ‰ salinities, showing a U-shaped curve relationship with the basal level in the 40 ‰ group. Hypothalamic PSS-I expression increased strongly in the 55 ‰ environment. GH mRNA levels did not change in any of the tested environmental salinities. PRL mRNA maximum levels were encountered in the 5 and 12 ‰ environments, but significantly down-regulated in the 40 ‰. Plasma cortisol levels significantly increased in the 40 ‰ environment. These results are discussed in relation to the well-known high adaptability of Sparus aurata to different environmental salinities and the role of the GH/IGF-I axis in this process.

GH, IGF-I and GH receptors mRNA expression in response to growth impairment following a food deprivation period in individually housed cichlid fish Cichlasoma dimerus.

Cichlasoma dimerus is a social cichlid fish capable of growing at high rates under laboratory conditions, but knowledge on somatic growth regulation is still unclear. Growth hormone (GH)/insulin-like growth factor I (IGF-I) axis is the key regulator of somatic growth in vertebrates. Two types of growth hormone receptors have been described in teleost fish, named GH receptor type 1 (GHR1) and type 2 (GHR2). In addition, isoforms of these receptors lacking part of the intracellular region have been described. The aim of this study was to evaluate the somatic growth, liver histology and changes in the GH/IGF-I axis after 4 weeks of food deprivation in C. dimerus. Four-week fasted fish showed reductions in specific growth rates in body weight (p < 0.001) and standard length (p < 0.001). Additionally, the hepatosomatic index (p < 0.001) and hepatocyte area (p < 0.001) decreased in fasted fish, while no changes in glucose levels were detected in plasma. The starvation protocol failed to induce changes in GH mRNA levels in the pituitary and IGF-I mRNA levels in liver. In contrast, IGF-I mRNA levels in muscle decreased in fasted fish (p = 0.002). On the other hand, GHR2 (detected with primer sets designed over the extracellular and intracellular region) was upregulated by starvation both in liver and muscle (p < 0.05), while GHR1 remained unchanged. These results show that a fasting period reduced somatic growth both in length and body weight concomitantly with alterations on liver and muscle GHR2 and muscle IGF-I mRNA expression.

GH Dysfunction in Engrailed-2 Knockout Mice, a Model for Autism Spectrum Disorders

Insulin-like growth factor 1 (IGF-1) signaling promotes brain development and plasticity. Altered IGF-1 expression has been associated to autism spectrum disorders (ASD). IGF-1 levels were found increased in the blood and decreased in the cerebrospinal fluid of ASD children. Accordingly, IGF-1 treatment can rescue behavioral deficits in mouse models of ASD, and IGF-1 trials have been proposed for ASD children. IGF-1 is mainly synthesized in the liver, and its synthesis is dependent on growth hormone (GH) produced in the pituitary gland. GH also modulates cognitive functions, and altered levels of GH have been detected in ASD patients. Here, we analyzed the expression of GH, IGF-1, their receptors, and regulatory hormones in the neuroendocrine system of adult male mice lacking the homeobox transcription factor Engrailed-2 (En2−/− mice). En2−/− mice display ASD-like behaviors (social interactions, defective spatial learning, increased seizure susceptibility) accompanied by relevant neuropathological changes (loss of cerebellar and forebrain inhibitory neurons). Recent studies showed that En2 modulates IGF-1 activity during postnatal cerebellar development. We found that GH mRNA expression was markedly deregulated throughout the neuroendocrine axis in En2−/− mice, as compared to wild-type controls. In mutant mice, GH mRNA levels were significantly increased in the pituitary gland, blood, and liver, whereas decreased levels were detected in the hippocampus. These changes were paralleled by decreased levels of GH protein in the hippocampus but not other tissues of En2−/− mice. IGF-1 mRNA was significantly up-regulated in the liver and down-regulated in the En2−/− hippocampus, but no differences were detected in the levels of IGF-1 protein between the two genotypes. Our data strengthen the notion that altered GH levels in the hippocampus may be involved in learning disabilities associated to ASD.

Dioxin-induced fetal growth retardation: the role of a preceding attenuation in the circulating level of glucocorticoid.

Exposure of pregnant rats to 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) at a low dose causes developmental disorders such as growth retardation and sexual immaturity in their pups. Our previous studies have demonstrated that TCDD attenuates the expression of pituitary luteinizing hormone in fetuses, resulting in the impairment of sexual behavior after they reach maturity. In this study, we focused on growth disturbance and investigated whether TCDD affects the expression of growth hormone (GH), another pituitary hormone which is essential for normal development in perinatal pups. The result showed that maternal exposure to TCDD (1 µg/kg) at gestational day (GD) 15 reduced the fetal expression of GH from the onset at GD18. In accordance with this, TCDD attenuated the pup weight during the perinatal period. We then examined the effect of TCDD on the serum concentration of corticosterone, which plays a key role in the proliferation of GH-producing cells, and found that TCDD reduces the circulating level of corticosterone in the mothers at GD18 and the male fetuses at GD19. The reduction in fetuses seems to be due to increased inactivation rather than reduced synthesis, because TCDD induces the fetal expression of hepatic enzymes participating in the metabolism of glucocorticoids without changing the expression of steroidogenic proteins in the pituitary-adrenal axis. Supplying corticosterone to TCDD-exposed mothers restored or tended to restore a TCDD-induced reduction in pup weight as well as the levels of pituitary GH mRNA and serum GH. These results suggest that TCDD lowers GH expression and growth retardation owing, at least partially, to a reduction in the circulating level of glucocorticoid in pregnant mothers and their fetuses.

Enhancement of transfection efficiency with NLS and SPB-NLS

Low positive cell screening efficiency severely hinders the development of transgenic animals. The major rate-limiting step of positive cell screening is DNA entering the nucleus, particularly for large DNA molecules. To enhance the transport of large DNA molecules into the nucleus, particularly for the production of transgenic animals, nuclear localization sequence (NLS) peptides and the peptide derivative succinimidyl-[4-(psoralen-8-yloxy)]-butyrate (SPB)-NLS were synthesized to mediate transfection in vitro. To investigate the function of NLS and SPB-NLS in vitro, the expression levels of growth hormone (GH) mRNA and green fluorescent protein (GFP) protein were analyzed following transfection mediated by NLS and SPB-NLS. The results demonstrated that the expression of GH mRNA was significantly higher in the NLS (increased by 69%) and SPB-NLS (330%) groups than that in the liposome/pGN group. Similarly, GFP expression was found to be higher in the SPB-NLS group than that in the liposome group, while the expression in the NLS group was lower than that in the liposome group. Further analysis demonstrated that SPB-NLS enhanced the expression of insulin-like growth factor 1 in hard-to-transfect goat mammary epithelia cells. The results of the microscopy analysis revealed that transfected DNA entered the nucleus via the nuclear pores, facilitated by NLS. Analysis of the cell cycle demonstrated that the cytotoxic effects of NLS and SPB-NLS were low. In conclusion, the results of the present study demonstrate that SPB-NLS acts as a transfection-enhancing agent and may be used both to enhance nuclear delivery and for the development of genetically modified animals.

Differential effects of exogenous and autocrine growth hormone on LNCaP prostate cancer cell proliferation and survival

The prostate gland is regulated by multiple hormones and growth factors that may also affect prostate tumorigenesis. Growth hormone (GH) contributes to prostate development and function, but the direct effects of GH on prostate cancer cells are not well understood. The expression of endogenous GH in prostate cancer cell lines has also been observed, suggesting the potential for an effect of autocrine GH. In the present study, we measure the levels of GH and GH receptor (GHR) mRNA in multiple prostate cancer and normal prostate-derived cell lines, and compare the effects of exogenous and autocrine GH on LNCaP prostate cancer cell proliferation and apoptosis, and the associated signal transduction pathways. We found that GHR and GH expression were higher in the prostate cancer cell lines, and that exogenous GH increased LNCaP cell proliferation, but had no effect on apoptosis. In contrast, autocrine GH overexpression reduced LNCaP cell proliferation and increased apoptosis. The distinct actions of exogenous and autocrine GH were accompanied by differences in the involvement of GHR-associated signal transduction pathways, and were paralleled by an alteration in the subcellular localization of GHR, in which autocrine GH appeared to sequester GHR in the Golgi and endoplasmic reticulum. This alteration of GHR trafficking may underlie a distinct mode of GH-mediated signaling associated with the effect of autocrine GH. These findings clarify the potential effects of GH on prostate cancer cell function, and indicate that the activity of autocrine GH may be distinct from that of endocrine GH in prostate cancer cells.

Glucocorticoid-induced changes in gene expression in embryonic anterior pituitary cells

Within the anterior pituitary gland, glucocorticoids such as corticosterone (CORT) provide negative feedback to inhibit adrenocorticotropic hormone secretion and act to regulate production of other hormones including growth hormone (GH). The ontogeny of GH production during chicken embryonic and rat fetal development is controlled by glucocorticoids. The present study was conducted to characterize effects of glucocorticoids on gene expression within embryonic pituitary cells and to identify genes that are rapidly and directly regulated by glucocorticoids. Chicken embryonic pituitary cells were cultured with CORT for 1.5, 3, 6, 12, and 24 h in the absence and presence of cycloheximide (CHX) to inhibit protein synthesis. RNA was analyzed with custom microarrays containing 14,053 chicken cDNAs, and results for selected genes were confirmed by quantitative reverse transcription real-time PCR (qRT-PCR). Levels of GH mRNA were maximally induced by 6 h of CORT treatment, and this response was blocked by CHX. Expression of 396 genes was affected by CORT, and of these, mRNA levels for 46 genes were induced or repressed within 6 h. Pathway analysis of genes regulated by CORT in the absence of CHX revealed networks of genes associated with endocrine system development and cellular development. Eleven genes that were induced within 6 h in the absence and presence of CHX were identified, and eight were confirmed by qRT-PCR. The expression profiles and canonical pathways defined in this study will be useful for future analyses of glucocorticoid action and regulation of pituitary function.

Neuronostatin alters gene expression in rat anterior pituitary cells: a possible role in energy balance and glucose homeostasis

Neuronostatin is a recently discovered peptide hormone derived from the somatostain preprohormone (JBC 287:4572, 2008). We now have identified that the orphan G protein‐coupled receptor, GPR107, is a potential cognate receptor for neuronostatin (AJP 303:R941, 2012). Because of the abundant expression of GPR107 in anterior pituitary cells, and because neuronostatin is present in somatostatin‐producing neurons of the hypothalamus that project to median eminence, we hypothesized that neuronostatin would alter hormone production in anterior pituitary. The ability of neuronostatin to activate early gene expression in pituitary gland further suggested trophic effects; however, we were initially unable to detect acute effects of the peptide on basal, stimulated or inhibited hormone secretion in cells in culture. Therefore, anterior pituitary cells were isolated from male rats, treated for longer time periods with neuronostatin, and changes in hormone gene expression were examined using real‐time polymerase chain reaction. We found that neuronostatin activated early gene expression and induced an increase in growth hormone mRNA levels, suggesting that neuronostatin may regulate growth hormone production, which could have important implications for energy homeostasis and glucose metabolism in addition to possible direct effects in the pancreas where the peptide also activates early gene expression. (NIH HL066023)

In vitro Suppression of Prolactin During Later Stages of Egg Lay in Domestic Hen (Gallus gallus Domesticus) Anterior Pituitcytes by RNA Interference

2 Abstract: Prolactin (PRL) is a peptide hormone secreted by anterior pituitary gland. PRL increases as per the age and reproductive cycle of birds from normal physiological levels to extremely higher level there by affecting the ovulation, egg formation, oviposition, egg production and hyperprolactinemia. This is mor e pronounced and persistent after 72 weeks of age in birds. Hence a study was conducted in anterior pituitary primary cultured cells obtained from 72 and 82 weeks old white leg horn (WLH) hens to knock down the PRL gene expression by siRNA and observing its effects on PRL, PRL mRNA, protein content of PRL, prolactin receptor (PRLR) and Growth Hormone (GH) mRNA to unravel the functional role of PRL at 72 and 82 week age by siRNA for PRL. Three siRNAs were designed as per standard siRNA protocols and studied the suppression of PRL gene expression in primary cultured cells procured from adult chicken anterior pituitary glands in in vitro conditions. Average percentage reduction of PRL in anterior pituitary primary cell culture following siRNA transfection was 82 and 60% at 72 and 82 weeks respectively. Protein content of PRL was significantly (p<0.01) decreased in siRNA transfected cells compared to controls. Growth Hormone (GH) mRNA and PRL receptor (PRLR) mRNA levels did not change significantly (p<0.01) between control and treated cells. Results clearly suggested that the siRNA designed for PRL specifically decreased PRL gene expression in in vitro conditions. Level of PRLR mRNA and GH mRNA levels expression did not follow the similar pattern of PRL gene expression in anterior pituitary cells. It is concluded that, construction of short specific siRNA for PRL significantly decreased PRL, PRL mRNA and protein content of PRL without showing any effect on PRLR and GH between the two age groups of birds. These results may lead to construction of short specific siRNA for stable and chronic suppression of PRL gene expression during embryogenesis before an increase of PRL occurs for long term knock down of PRL in in vivo conditions. In conclusion , understanding of hyperprolactinemia and the involvement of PRL may provide the basis for the development of therapeutic drugs or methods against hyperprolactinemia by RNA interference.

Insulin-Like Growth Factor 1 Mediates Negative Feedback to Somatotroph GH Expression via POU1F1/CREB Binding Protein Interactions

Circulating insulin-like growth factor 1 (IGF-1) has been shown to act as a negative feedback regulator of growth hormone (GH) gene expression; however, the mechanism of this negative feedback is poorly understood. Activation and regulation of GH gene expression require the binding of the transcription factor POU1F1 to the GH promoter along with cyclic AMP (cAMP) response element binding protein (CREB) binding protein (CBP). We investigate the role of CBP as a target of IGF-1 somatotroph regulation using the MtT/S somatotroph cell line. IGF-1 significantly inhibits basal GH mRNA levels but not POU1F1 levels. Chromatin immunoprecipitation assays demonstrate inhibition of CBP binding to the GH promoter after IGF-1 treatment. We hypothesized that IGF-1 receptor (IGF-1R) signaling disrupts the POU1F1/CBP complex to inhibit gene expression. In support, the use of a mutant CBP (S436A) construct, which lacks a critical phosphorylation site, leads to the loss of IGF-1 inhibition. The studies of CBP (S436A) knock-in mice show elevated serum GH levels, a greater response to GH releasing hormone (GHRH) stimulation along with lower weight gain, and decreased body fat. Our data confirm the inhibitory effects of IGF-1 on GH expression at the level of the promoter and provide evidence of CBP's role as a target of IGF-1R signaling.

Effects of cysteamine on mRNA levels of growth hormone and its receptors and growth in orange-spotted grouper (Epinephelus coioides)

Effects of cysteamine (CS) on growth hormone (GH) mRNA, two types of growth hormone receptor (GHR) mRNAs and growth rate in orange-spotted grouper (Epinephelus coioides) were investigated. CS could cause a modification in the structure of somatostatin, which is the most important neuroendocrine inhibitor of basal and stimulated growth hormone synthesis and release, and renders it nonimmunoreactive probably through interaction with the disulfide bonds. In the present study, cysteamine hydrochloride (CSH) enhanced the level of pituitary GH mRNA in a dose-dependent manner through attenuating or deleting the inhibiting action of somatostatin on GH mRNA expression. CSH at relatively low doses (from 1 to 3mg/g diet) enhanced the levels of two types of GHR mRNAs in dose-dependent manner, whereas the stimulation induced by CSH declined from the peak at higher dose of CSH (4mg/g diet). It might be attributed to the variation in GH-induced up-regulation of GHRs at different doses of GH. Feeding of CSH could induce remarkable enhancement of growth rate in orange-spotted grouper. In addition, the stimulatory effect of CSH could be potentiated by the additive effect of luteinizing hormone-releasing hormone analog (LHRH-A). Compared with individual treatments, combined feeding of CSH and LHRH-A caused more efficient elevation of growth rate after 8weeks of feeding. CSH and LHRH-A individually and in combination remarkably increased the levels of GH and GHR mRNAs compared with the control. The combined administration of CSH and LHRH-A in diet was most effective to enhance the level of GH and GHR1 mRNA. The morphological characteristics of the experimental fish were evaluated. Compared with control, the ratios of muscle RNA/DNA, condition factors (CF) and feed conversion efficiency (FCE) were significantly enhanced in the treated groups, while the highest values were observed in the combined treatment. All the results suggested that CSH (1-3mg/g diet) is an effective, economical and feasible feed additive in orange-spotted grouper culture.

Stimulatory effects of neuropeptide Y on the growth of orange-spotted grouper (Epinephelus coioides)

Neuropeptide Y (NPY) is a member of the pancreatic polypeptide family which is a potent orexigenic peptide known to date in mammals and teleost. This study was carried out to investigate the effects of NPY on food intake and growth of orange-spotted grouper (Epinephelus coioides). Synthetic grouper NPY (gNPY) was given orally at the dose of 0.5, 1.0 and 2.0μg/g feed for 50days, results showed that NPY treatment (1.0 and 2.0μg/g feed) significantly increased growth rate, weight gain, feed conversion efficiency (FCE) and pituitary growth hormone (GH) mRNA level than the control group (p<0.05). Furthermore, high level secretion of gNPY was expressed and purified in the Pichia pastoris expression system. The bioactivity of recombinant gNPY was confirmed by its ability to up-regulate GH mRNA expression in vivo and in vitro and down-regulate preprosomatostatin I (PSSI) mRNA expression in vivo. These results demonstrate that NPY has stimulatory effects on food intake as well as growth of grouper as in other teleost fish, also indicate that recombinant gNPY from P. pastoris has the same bioactivity as synthetic gNPY and has the potential to be used as a feed additive for both research and aquatic application.

Direct effects of triiodothyronine on production of anterior pituitary hormones and gonadal steroids in goldfish

The present study investigated the effects of triiodothyronine (T3) on pituitary gonadotropin (GTH) subunits, thyroid stimulating hormone (TSH) β subunit, and growth hormone (GH) mRNA levels, as well as gonadal steroid secretion during different stages of reproduction in goldfish. Goldfish pituitary cells cultured with T3 exhibited lower tshβ mRNA levels in all reproductive stages and lower luteinising hormone β (lhβ) mRNA levels in early recrudescence, whereas gh and fshβ mRNA levels were not altered. T3 injections significantly reduced circulating oestrogen (OE2) concentrations in early and mid recrudescent male goldfish, but were without effect on the circulating level of OE2 in female fish. T3 injections also reduced circulating levels of testosterone in both male and female goldfish during the mid stage of gonadal recrudescence. In vitro culture of goldfish ovarian follicles at the late stage of gonadal recrudescence, in the presence of T3, resulted in reduced OE2 secretion; no consistent effect of T3 on testosterone secretion was observed in cultured goldfish ovarian follicles and testis. These findings support the hypothesis that T3 impairs reproduction by inhibiting production of gonadal steroids and pituitary luteinising hormone production in goldfish.

High Lever Dietary Copper Promote Ghrelin Gene Expression in the Fundic Gland of Growing Pigs

This experiment was conducted to examine the effect of dietary copper supplementation on ghrelin mRNA expression level in the fundic gland of growing pigs. A total of 45 crossbred pigs were randomly assigned to three groups of 15 pigs, five replicates of three animals comprised each group. Pigs were allocated to diets that contained 5mg/kg copper (as the control group), 125mg/kg copper sulfate, or 125mg/kg copper methionine. At the end of the experiment, five pigs were selected at random from each group, slaughtered, and collected the fundic gland for determination of ghrelin mRNA expression level. The results showed that average daily gain, average daily feed intake, absolute weight, serum growth hormone (GH) concentration, and ghrelin mRNA level were higher in pigs fed the diets with 125mg/kg copper methionine and 125mg/kg copper sulfate (P < 0.05), than in pigs fed a diet with 5mg/kg copper. These data suggest that high dietary copper (125mg/kg) appears to increase feed intake and promote weight gain by enhancing the secretion of GH and ghrelin mRNA level in growing pigs.