Growth Hormone mRNA Research Articles

Aryl‐hydrocarbon receptor activity modulates prolactin expression in the pituitary

Pituitary tumors account for 15% of intracranial neoplasms, however the extent to which environmental toxicants contribute to the proliferation and hormone expression of pituitary cells is unknown. Aryl-hydrocarbon receptor (AhR) interacting protein (AIP) loss of function mutations cause somatotrope and lactotrope adenomas in humans. AIP sequesters AhR and inhibits its transcriptional function. Because of the link between AIP and pituitary tumors, we hypothesize that exposure to dioxins, potent exogenous ligands for AhR that are persistent in the environment, may predispose to pituitary dysfunction through activation of AhR. In the present study, we examined the effect of AhR activation on proliferation and endogenous pituitary hormone expression in the GH3 rat somatolactotrope tumor cell line and the effect of loss of AhR action in knockout mice. GH3 cells respond to nM doses of the reversible AhR agonist β-naphthoflavone with a robust induction of Cyp1a1. Although mRNA levels of the anti-proliferative signaling cytokine TGFbeta1 are suppressed upon β-naphthoflavone treatment, we did not observe an alteration in cell proliferation. AhR activation with β-naphthoflavone suppresses Ahr expression and impairs expression of prolactin (PRL), but not growth hormone (GH) mRNA in GH3 cells. In mice, loss of Ahr similarly leads to a reduction in Prl mRNA at P3, while Gh is unaffected. Additionally, there is a significant reduction in pituitary hormones Lhb and Fshb in the absence of Ahr. Overall, these results demonstrate that AhR is important for pituitary hormone expression and suggest that environmental dioxins can exert endocrine disrupting effects at the pituitary.

Interactive effects of temperature-dietary protein level on somatotropic gene expression and its interrelationship with growth in juvenile GIFT tilapia Oreochromis niloticus

Water temperature and dietary protein play an important role in influencing the expression of somatotropic gene. Based on central composite rotatable experimental design and response surface method, the interacting effects of temperature (20–34°C) and dietary protein level (25–50%) on expression of somatotropic gene and its interrelationship with specific growth rates (SGR) and feed efficiency (FE) of juvenile GIFT tilapia were studied under laboratory conditions. Results showed that the linear, quadratic and interacting effects of temperature and dietary protein on both pituitary growth hormone (GH) and hepatic growth hormone receptor 1 (GHR1) mRNA levels were statistically significant (P<0.05). The linear and interacting effects of temperature and dietary protein on hepatic insulin-like growth factor I (IGF-I) mRNA levels were highly significant (P<0.01); but the quadratic effect of temperature was nonsignificant (P>0.05). The regression equations of pituitary GH, hepatic GHR1 and IGF-I mRNA towards the two factors of interest were established, with the coefficients of determination being 0.958, 0.938 and 0.978, respectively (P<0.01). Pituitary GH, hepatic GHR1 and IGF-I showed positive relationships with SGR and FE. These results suggest that the expression of somatotropic gene be affected by different combination of temperature and dietary protein level as they regulate growth and FE.

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.

The effect of glycerol on mRNA expression of growth hormone, insulin-like growth factor, and mitochondrial breast muscle genes of Japanese quail

1. A study was carried out to evaluate the expression of growth hormone (GH), insulin-like growth factor I (IGF-I), mitochondrial adenine nucleotide translocase (ANT), cytochrome oxidase III (COX III) and avian uncoupling protein (avUCP) genes in 14-d-old Japanese quail that were fed different levels of glycerol (0%, 4%, and 12% dietary glycerol) which replaced maize as an energy source. 2. Total RNA was extracted from the breast muscle, and cDNA was amplified using real-time PCR with primers specific to the examined genes. 3. Quail fed the diet with 12% glycerol supplementation presented higher growth hormone (GH) mRNA expression than did those fed 0% glycerol. Supplementation with 12% glycerol negatively influenced IGF-I mRNA expression and reduced ANT mRNA expression in comparison with the treatment with no glycerol. COX III mRNA expression in the pectoralis superficialis muscle was decreased by 26% in quail fed 12% glycerol compared with those fed 0 and 4% glycerol. 4. There was no difference in UCP mRNA expression between quail fed 0 and 4% glycerol; however, UCP expression was reduced (73%) in birds fed the 12% glycerol diet compared with the level in those fed the 4% glycerol diet. 5. The inclusion of 4% glycerol in the diet produced results similar to those of the diet with no glycerol. Based on quail performance and the expression of the GH, IGF-I, ANT, COX III and UCP genes, 4% glycerol can be used in quail feeding without any harmful effects.

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.

Molecular characterization of the GHRH/GHRH-R and its effect on GH synthesis and release in orange-spotted grouper (Epinephelus coioides)

Growth hormone-releasing hormone (GHRH) is a hypothalamic neuropeptide that stimulates growth hormone (GH) synthesis and secretion in the pituitary gland. In this paper, the full-length cDNAs of orange-spotted grouper GHRH and its receptor (GHRH-R) were cloned. The grouper GHRH cDNA is 713 bp in length and encodes a 141-aa precursor that includes an 18-aa signal peptide, a 27-aa mature GHRH mature peptide and a 47-aa carboxyl terminus. The grouper GHRH-R cDNA sequence is 1495 bp in length, encoding a 422-aa receptor with seven transmembrane domains. Tissue distribution analyses showed that both GHRH and GHRH-R mRNAs were predominantly expressed in the brain, while the GHRH-R mRNA was also abundantly detected in the pituitary gland. Both GHRH and GHRH-R mRNAs were expressed throughout embryonic development from the multi-cell stage to the newly hatched larvae stage, and the highest GHRH and GHRH-R expressions appeared at the brain vesicle stage and the heart stage, respectively. In vitro studies performed on the grouper pituitary primary cells showed that a synthetic grouper GHRH-NH(2) increased both GH mRNA expression and GH protein release in a dose-dependent manner. Together, these results suggest that the newly obtained grouper GHRH was able to stimulate GH synthesis and release, similar to its mammalian counterparts.

Elevated expressions of GH/IGF axis genes in triploid crucian carp

Growth hormone (GH), growth hormone receptor (GHR) and insulin-like growth factor 1 (IGF-1) are pivotal signaling factors of the GH/IGF axis, which plays a crucial role in regulating growth in vertebrates. In this study, GH, GHR and IGF-1 cDNAs were cloned from triploid and tetraploid crucian carp. In addition, mRNA expression levels were characterized in diploid red crucian carp, triploids and tetraploids. Reverse transcriptase PCR indicated that GH genes were only expressed in the pituitary, while GHR and IGF-1 were widely expressed in all tested tissues. Real-time PCR study of different seasonal profiles showed that triploids had significantly higher expression of the studied genes during both the prespawning and the spawning season. Although different temperatures (22, 26 and 30°C) showed no significant effects on GH, GHR and IGF-1 mRNA expression in either diploids or triploids, triploids had higher expression levels than diploids at each temperature. After 1week of fasting, the expression of all studied genes was reduced in both diploids and triploids, while the expressions levels were higher in triploids than in diploids. These results suggest that the elevated expression of GH/IGF axis genes in triploids plays a crucial role in the faster growth rate of triploids.

Arginine induces GH gene expression by activating NOS/NO signaling in rat isolated hemi-pituitaries

The amino acid arginine (Arg) is a recognized secretagogue of growth hormone (GH), and has been shown to induce GH gene expression. Arg is the natural precursor of nitric oxide (NO), which is known to mediate many of the effects of Arg, such as GH secretion. Arg was also shown to increase calcium influx in pituitary cells, which might contribute to its effects on GH secretion. Although the mechanisms involved in the effects of Arg on GH secretion are well established, little is known about them regarding the control of GH gene expression. We investigated whether the NO pathway and/or calcium are involved in the effects of Arg on GH gene expression in rat isolated pituitaries. To this end, pituitaries from approximately 170 male Wistar rats (∼250 g) were removed, divided into two halves, pooled (three hemi-pituitaries) and incubated or not with Arg, as well as with different pharmacological agents. Arg (71 mM), the NO donor sodium nitroprusside (SNP, 1 and 0.1 mM) and a cyclic guanosine monophosphate (cGMP) analogue (8-Br-cGMP, 1 mM) increased GH mRNA expression 60 min later. The NO acceptor hemoglobin (0.3 µM) blunted the effect of SNP, and the combined treatment with Arg and L-NAME (an NO synthase (NOS) inhibitor, 55 mM) abolished the stimulatory effect of Arg on GH gene expression. The calcium channel inhibitor nifedipine (3 µM) also abolished Arg-induced GH gene expression. The present study shows that Arg directly induces GH gene expression in hemi-pituitaries isolated from rats, excluding interference from somatostatinergic neurons, which are supposed to be inhibited by Arg. Moreover, the data demonstrate that the NOS/NO signaling pathway and calcium mediate the Arg effects on GH gene expression.

Pit-1w may regulate prolactin gene expression in mouse testis

Pit-1 is a POU-domain transcription factor that promotes growth hormone (GH), prolactin (PRL), and thyroid-stimulating hormone β subunit (TSHβ) gene expression in the pituitary gland. Alternative splicing of Pit-1 gene transcripts has been shown to give rise to several variants with discrete transactivation properties. Recently, we identified a mouse Pit-1w that is generated by alternative promoter usage and is expressed in a variety of tissues including the testis. Using a combination of reverse-transcription polymerase chain reaction analyses and luciferase reporter gene assays, we investigated the possible role of Pit-1w in the mouse testis. In postnatal testicular development, the expression of Pit-1w mRNA was significantly up-regulated between 18 and 20days after birth when the numbers of secondary spermatocytes and spermatids have been reported to increase in mice. The PRL mRNA, but not the mRNAs for GH or TSHβ, showed intratesticular expression patterns that were similar to those of the Pit-1w mRNA. In experimental unilaterally cryptorchid testes of adult mice, spermatid numbers were extremely low and the expression levels of both the Pit-1w and PRL mRNAs dropped dramatically. Furthermore, in the luciferase reporter gene assays, we found that Pit-1w specifically transactivated the PRL promoter but had no effect on the promoters of GH or TSHβ. These results suggested that Pit-1w could be involved in the paracrine/autocrine system in mice and may be necessary for normal testicular function via its possible role in regulating PRL expression in testicular germ cells. This is the first report demonstrating the possible role of Pit-1w in mammals.

Growth and endocrine effects of recombinant bovine growth hormone treatment in non-transgenic and growth hormone transgenic coho salmon

To examine the relative growth, endocrine, and gene expression effects of growth hormone (GH) transgenesis vs. GH protein treatment, wild-type non-transgenic and GH transgenic coho salmon were treated with a sustained-release formulation of recombinant bovine GH (bGH; Posilac™). Fish size, specific growth rate (SGR), and condition factor (CF) were monitored for 14weeks, after which endocrine parameters were measured. Transgenic fish had much higher growth, SGR and CF than non-transgenic fish, and bGH injection significantly increased weight and SGR in non-transgenic but not transgenic fish. Plasma salmon GH concentrations decreased with bGH treatment in non-transgenic but not in transgenic fish where levels were similar to controls. Higher GH mRNA levels were detected in transgenic muscle and liver but no differences were observed in GH receptor (GHR) mRNA levels. In non-transgenic pituitary, GH and GHR mRNA levels per mg pituitary decreased with bGH dose to levels seen in transgenic salmon. Plasma IGF-I was elevated with bGH dose only in non-transgenic fish, while transgenic fish maintained an elevated level of IGF-I with or without bGH treatment. A similar trend was seen for liver IGF-I mRNA levels. Thus, bGH treatment increased fish growth and influenced feedback on endocrine parameters in non-transgenic but not in transgenic fish. A lack of further growth stimulation of GH transgenic fish suggests that these fish are experiencing maximal growth stimulation via GH pathways.

Effects of LED light spectra on the growth of the yellowtail clownfish Amphiprion clarkii

Growth hormone (GH) is an essential polypeptide required for the normal growth and development of vertebrates. We have studied the effects of light-emitting diodes (LEDs) emitting different spectra (red, green, and blue) on the GH of yellowtail clownfish Amphiprion clarkii. Full-length GH cDNA from the pituitary of the yellowtail clownfish was first cloned and then the expression of GH mRNA under different light spectra was measured. GH mRNA expression was significantly higher under green and blue light than under red light spectra. These results indicate that in yellowtail clownfish, short-wavelength LED enhances growth more than long-wavelength LED, and that LED lights are more effective for enhancing growth than white fluorescent bulbs. Injection of melatonin resulted in significantly higher expression levels of GH mRNA compared to the control. We therefore conclude that green and blue light enhance GH levels and that melatonin plays a role in modulating growth of the yellowtail clownfish.

Effects of Polybrominated Diphenyl Ethers (PBDEs) and Their Derivatives on Protein Disulfide Isomerase Activity and Growth Hormone Release of GH3 Cells

Polybrominated diphenyl ethers (PBDEs) have been used in a variety of consumer products such as flame retardants and recently have been known to be widespread environmental pollutants, which probably affect biological functions of mammalian cells. However, the risk posed by PBDE metabolites has not been clarified. Our previous study suggested that bisphenol A (BPA), an endocrine-disrupting chemical, binds to protein disulfide isomerase (PDI) and inhibits its activity. PDI is an isomerase enzyme in the endoplasmic reticulum and facilitates the formation or cleavage of disulfide bonds. PDI consists of a, b, b', and a' domains and the c region, with the a and a' domains having isomerase active sites. In the present study, we tested the effects of 10 kinds of PBDE compounds and their metabolites on PDI. OH-PBDEs specifically inhibited the isomerase activity of PDI, with 4'-OH-PBDE more effective than 2' (or 2)-OH-PBDEs. 4'-OH-PBDE inhibited the isomerase activity of the b'a'c fragment but not that of ab and a'c, suggesting that the b' domain of PDI is essential for the inhibition by 4'-OH-PBDE. We also investigated the effects of these chemicals on the production of growth hormone (GH) in GH3 cells. In GH3 cells, levels of mRNA and protein of GH stimulated by T(3) were reduced by 4'-OH-PBDE and 4'-MeO-PBDE. The reduction in GH expression caused by these compounds was not changed by the overexpression or knockdown of PDI in GH3 cells, while these manipulations of PDI levels significantly suppressed the expression of GH. These results suggest that the biological effects of PBDEs differed depending on their brominated and hydroxylated positions.

Release of retinal growth hormone in the chick embryo: Local regulation?

The neural retina is an extrapituitary site of growth hormone (GH) production and an autocrine or paracrine site of retinal GH action. Retinal GH is released from retinal tissue and may be secreted into the vitreous. Ontogenetic changes in the abundance of retinal GH during embryogenesis indicate that the amount of GH released may be regulated. The presence of pituitary GH secretagogues (GH-releasing hormone, GHRH; thyrotropin-releasing hormone, TRH; and ghrelin) and pituitary GH inhibitors (somatostatin, SRIF and insulin-like growth factor, IGF-1) within the neural retina may indicate the involvement of these factors in retinal GH release. This possibility is supported by the finding that GHRH is colocalized with GH in chick retinal ganglion cells (RGCs) and in immortalized cells (QNRD) derived from quail neuroretinal cells and by the induction of GH mRNA in incubated QNRD cells. In summary, these results provide evidence for the autocrine or paracrine regulation of retinal GH release in the ganglion cells of the embryonic chick retina.

Genetically modified growth affects allometry of eye and brain in salmonids

Effects of growth acceleration on eye development have been examined in genetically modified salmonids. Growth hormone (GH) transgenic coho salmon ( Oncorhynchus kisutch (Walbaum, 1792)) show dramatically elevated overall body growth and an absolute increase in eye size, but relative eye growth is shifted from negatively allometric to more isometric. Thus, transgenic fish possess significantly smaller eyes relative to nontransgenic fish of the same size. Ration-restricted limitation of growth in transgenic salmon to that of wild type restores relative eye growth rate, suggesting that effects on eyes are an indirect consequence of modification of growth rate rather than a direct effect of GH overexpression. Heart, spleen, and liver did not show changes in proportion among groups, whereas total brain size showed the same response as eye. Relative eye and brain size were also reduced in a fast-growing domesticated strain of rainbow trout ( Oncorhynchus mykiss (Walbaum, 1792)), suggesting modifications of allometry are a more general effect of growth acceleration. GH mRNA levels from the transgene were elevated in eyes, whereas IGF-I mRNA was not, suggesting this organ may be regulated in a different fashion than other organs. Neural tissues with critical structural requirements for optimal function may be subject to less modification of growth rate than are other organ systems.

Epidermal Growth Factor‐Activated Extracellular Signal‐Regulated Kinase Suppresses Growth Hormone Expression and Stimulates Proliferation in MtT/ E Cells

The mechanism for the inhibition of growth hormone (GH) expression by the epidermal growth factor (EGF) was examined in two clonal cell lines, MtT/E and MtT/S. The former has a negligible basal level of GH, whereas the latter has a high basal GH. The treatment of MtT/E cells with retinoic acid resulted in a significant increase in GH mRNA and subsequently GH. This stimulatory response to retinoic acid was strongly suppressed by EGF. This suppression was associated with an increase in the phosphorylation of extracellular signal-regulated kinase 1 and 2 (Erk1/2). The MEK [mitogen-activated protein kinase (MAPK) kinases that activate ERK1 and ERK2] inhibitor, PD98059, clearly inhibited the phosphorylation of Erk1/2 and restored the stimulatory effects of retinoic acid. These results suggest that the inhibitory effects of EGF on GH expression are mediated by MAPK activation in these cells. By contrast to the GH-producing clones examined previously, EGF showed a marked stimulation of proliferation of the MtT/E cells through a mechanism dependent on MAPK activation. On the other hand, the inhibitory effect of EGF on GH expression was less pronounced and the stimulation of cellular proliferation was not seen in MtT/S cells, even though it induced Erk-phosphorylation similar to that seen in MtT/E. The distinct difference in the response to EGF between these two GH cell lines appears to be attributed to differences in the function of MAPK cascade in each cell line. This may reflect the developmental stage of the cells from which MtT/E and MtT/S are derived.

Regulation of somatic growth and gene expression of the GH–IGF system and PRP-PACAP by dietary lipid level in early juveniles of a teleost fish, the pejerrey (Odontesthes bonariensis)

Growth and mRNA levels of the pituitary adenylate cyclase-activating polypeptide (PACAP) and its related peptide (PRP), and the system controlled by the growth hormone (GH) and insulin-like growth factors (IGFs) were analyzed in pejerrey fry fed with graded levels of dietary lipids: 10% (L10), 13% (L13) and 21% (L21). First, the full sequence of pejerrey PRP-PACAP was obtained by RT-PCR, using primers based on conserved fragments of teleosts PACAP sequences. The growth of the fish at 83days after hatching (dah) and the GH mRNA levels were not significantly affected by the dietary treatment. Conversely, PRP-PACAP expression significantly decreased with increasing dietary lipids (L10>L21). While GH receptor (GHR)-I and IGF-I transcripts did not differ among groups, GHR-II transcripts decreased in group L21. IGF-II expression apparently followed the same trend. These results in combination with the lower expression of the anorexigenic PRP-PACAP in fish fed diet L21 and the correlation analysis evidencing a particularly fine tuning of the GH-IGF system in group L13, suggest that this diet may cover the energy demands for growing pejerrey from 27dah onwards. Our results show for first time in fish a differential response of PRP-PACAP transcripts to dietary manipulations, and confirm the sensitivity of the pejerrey GH-IGF system to changes in diet composition despite the lack of (or in advance to) a clear response of somatic growth.

Artesunate inhibits cell proliferation and decreases growth hormone synthesis and secretion in GH3 cells

To determine the effect of artesunate (ART) on the rat pituitary adenoma GH3 cell line to evaluate its potential as a novel agent in growth hormone (GH) adenoma and to investigate its underlying mechanisms of action. The MTT assay was used to assess cell proliferation. DAPI staining was used to visualise apoptotic changes in the nucleus. We also analyzed cell apoptosis and cell cycle stage by flow cytometry, semi-quantitative RT-PCR analysis for the expression of GH mRNA and apoptosis-induced factor (AIF) mRNA, analysis of GH protein by western blot, ELISA detection of secreted GH, and the caspase inhibition assay. We found that ART inhibited the proliferation of GH3 cells in a dose- and time-dependent manner, with an IC50 of 9.53±4.12μM. The IC50s of ART against of two normal cell lines (mouse embryonic fibroblasts, and rat bone mesenchymal cells) were much higher than the IC50 recorded for the GH3 cells. ART induced apoptosis and blocked GH3 at G2/M arrest. The pan caspase inhibitor V-ZAD-FMK partly attenuated the inhibitory effect of ART. ART increased the expression of AIF mRNA and reduced GH mRNA levels, GH synthesis and the secretion of GH level in GH3 cells. ART can inhibit proliferation and induce apoptosis in GH3 cells by caspase-dependent pathways. Additionally, ART can inhibit GH synthesis and secretion. Thus, we propose ART as a probably anti-tumour candidate drug in the treatment of GH adenoma.

Cloning and differential expression of growth hormone inPelteobagrus vachelli

Growth hormone(GH) is a pluripotent hormone produced by the pituitary gland in teleosts as in other vertebrates.It plays important roles in the growth,development and metabolism of vertebrates.In this study,to characterize GH in fish and to better understand the potential applications of GH in fish cul-ture,we used reverse transcription-ploymerase chain reaction(RT-PCR) and rapid amplification of cDNA ends(RACE) to obtain the full-length cDNA sequence encoding GH from the pituitary of darkbarbel cat-fish Pelteobagrus vachelli(P.vachelli),a freshwater fish of high commercial value cultured in China.GH precursor cDNA(GenBank:GU395549) consists of 1 203 bp in size with an open-reading frame(ORF) encoding 200 amino acid,a 77 bp 5'-untranslated region and a 523 bp 3'-untranslated region.The four Cys residues are located at conserved positions(Cys71,Cys173,Cys190,Cys198),and one possible site for N-glycosylation(Asn-Xaa-Ser/Thr,motif) is present at Asn197.Based on multi-sequence alignments and phylogenetic analysis,the P.vachelli GH precursor shared a higher degree of homology(averaged 95%) with other species of fish belonging to the Order Siluriformes,but lower levels of homology between P.vachelli and the mammalian(averaged 34%) and bird(averaged 41%).The above results reveal that GH is conserved during evolution and is suitable for phylogenetic relationship analysis of Siluriformes.By real-time quantitative PCR,tissue distribution profile showed that GH mRNA signals were all observed in pituitary,brain,muscle,liver,adipose tissue,stomach,spleen,ovary and testis.GH mRNA in pituitary was the most abundant and no significant GH mRNA expression was detected in kidney,heart,gill and intestines.The expression patterns of P.vachelli and previously studied animal GH suggest that GH through autocrine or paracrine pathway exerted its possible important local physiological function during development and growth of P.vachelli.These findings will contribute to the understanding of the evolution of GH and provide some basic information about the characterization of GH in P.vachelli.

Expression of growth genes in response to glycerol use in Japanese quail diets

Glycerol can be used as a substitute for corn for feeding poultry, but there are concerns about how it may affect growth performance and health of the birds. We evaluated the expression of mRNA of growth hormone (GH) and insulin-like growth factor I (IGF-I) in 35-day-old Japanese quails fed different glycerol levels (0, 4, and 8% dietary glycerol instead of corn). Total RNA was extracted from the breast muscle and cDNA was amplified with the use of specific primers for these genes using real-time PCR. Quails fed the diet with 8% glycerol supplementation had significantly lower GH mRNA and IGF-I mRNA expression than those fed no glycerol or 4% glycerol. No significant effect of the treatments was found on quail weight gain or feed intake. Feed conversion ratio was influenced by dietary glycerol levels: the group fed 8% glycerol displayed the worst feed conversion ratio (2.54) compared with that of quail fed the control diet (2.35) or 4% glycerol (2.36). Considering quail performance and the expression of the genes GH and IGF-I, a level of 4% glycerol can be used in quail feeding without any harmful effects.