Endocrine effects of growth hormone overexpression in transgenic coho salmon (original) (raw)
Related papers
General and Comparative Endocrinology, 2012
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 14 weeks, 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 nontransgenic 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.
Transgenic Research, 2005
We have previously produced transgenic fish from crosses between a wild-type female tilapia (Oreochromis niloticus) and a G1 transgenic male. This line of growth-enhanced tilapia carries a single copy of a chinook salmon (s) growth hormone (GH) gene spliced to an ocean pout antifreeze promoter (OPA-FPcsGH) co-ligated to a carp b-actin/lacZ reporter gene construct, integrated into the tilapia genome. Because little is known about the expression sites of transgenes, we have characterised the gene expression patterns of sGH and tilapia (t)GH in transgenic tilapia using a newly established real-time PCR to measure the absolute mRNA amounts of both hormones. The sGH gene, which was expected to be expressed mainly in liver, was also found to be expressed in other organs, such as gills, heart, brain, skeletal muscle, kidney, spleen, intestine and testes. However, in pituitary no sGH mRNA but only tGH mRNA was found. Tilapia GH mRNA in wild-type pituitary amounted to 226 ± 30 pg/lg total RNA but in transgenics only to 187 ± 43 pg/lg total RNA. Liver exhibited the highest level of sGH mRNA (8.3 ± 2.5 pg/lg total RNA) but the extrahepatic sites expressed considerable amounts of sGH mRNA ranging from 4.1 ± 2.0 pg/lg total RNA in gills to 0.2 ± 0.08 pg/lg total RNA in kidney. The widespread expression of the sGH gene is assumed to be due to the tissue specificity of the type III AFP gene promoter. It is assumed that our transgenic experiments, which in contrast to some other approaches caused no obvious organ abnormalities, mimick the GH expression during ontogeny. Because sGH mRNA is expressed both in liver and in extrahepatic sites it may not only promote secretion and release of liver-derived (endocrine) IGF-I leading to an overall growth enhancement but also stimulate IGF-I expression within the different organs in a paracrine/autocrine manner and, thus, further promote organ growth.
Growth Hormone Endocrinology of Salmonids: Regulatory Mechanisms and Mode of Action
Fish Physiology and Biochemistry, 2000
The focus of this review is on the regulatory mechanisms and the mode of action of GH in salmonids. To stimulate further research, it aims at highlighting areas where numerous important breakthroughs have recently been made, as well as where data are currently lacking. The regulation of GH secretion is under complex hypothalamic control, as well as under negative feedback control by GH and IGF-I. Further, the recently characterized ghrelin is a potent GH secretagogue, and may prove to be a link between feed intake and growth regulation. GH plasma profiles show indications of diurnal changes, but whether salmonids have true pulsatile GH secretion remains to be elucidated. The recent cloning and characterization of the salmon GH receptor (GHR) is a major research break-through which will give new insights into the mechanisms of GH action. It should also stimulate research into circulating GH-binding proteins (GHBPs), as they appear to be a soluble form of the GHR. The salmonid GHR sequences show evolutionary divergence from other fish species, but with a high degree of identity within the salmonid group. Radioreceptorassay studies have found GHR present in all tissues examined, which is in line with the highly pleiotropic action of GH. Data are currently scarce on the plasma dynamics of GH in salmonids, and further studies on GHR and GHBPs dynamics coupled to assessments of GH clearance rates and pathways are needed. The direct versus indirect nature of GH action remains to be clarified, but GH appears to act both locally at the target tissue level to stimulate the autocrine/paracrine action of IGF-I, as well as on the liver to increase plasma IGF-I levels. In addition, GH interacts with other hormones such as cortisol, thyroid hormones, insulin, and reproductive hormones, generating a wide range of physiological effects. GH may act both peripherally and directly at the level of the central nervous system to modify behavior, probably by altering the dopaminergic activity in the brain.
Effect of growth hormone overexpression on gastric evacuation rate in coho salmon
Fish Physiology and Biochemistry, 2017
Growth hormone (GH) transgenic (T) coho salmon consistently show remarkably enhanced growth associated with increased appetite and food consumption compared to non-transgenic wild-type (NT) coho salmon. To improve understanding of the mechanism by which GH overexpression mediates food intake and digestion in T fish, feed intake and gastric evacuation rate (over 7 days) were measured in size-matched T and NT coho salmon. T fish displayed greatly enhanced feed intake levels (~2.5-fold), and more than 3-fold increase in gastric evacuation rates relative to NT coho salmon. Despite the differences in feed intake, no differences were noted in the time taken from first ingestion of food to stomach evacuation between genotypes. These results indicate that enhanced feed intake is coupled with an overall increased processing rate to enhance energy intake by T fish. To further investigate the molecular basis of these responses, we examined the messenger RNA (mRNA) levels of several genes in appetite-and gastric-regulation pathways (Agrp1, Bbs, Cart, Cck, Glp, Ghrelin, Grp, Leptin, Mc4r, Npy, and Pomc) by qPCR analyses in the brain (hypothalamus, preoptic area) and pituitary, and in peripheral tissues associated with digestion (liver, stomach, intestine, and adipose tissue). Significant increases in mRNA levels were found for Agrp1 in the preoptic area (POA) of the brain, and Grp and Pomc in pituitary for T coho salmon relative to NT. Mch and Npy showed significantly lower mRNA levels than NT fish in all brain tissues examined across all time-points after feeding. Mc4r and Cart for T showed significantly lower mRNA levels than NT in the POA and hypothalamus, respectively. In the case of peripheral tissues, T fish had lower mRNA levels of Glp and Leptin than NT fish in the intestine and adipose tissue, respectively. Grp, Cck, Bbs, Glp, and Leptin in stomach, adipose tissue, and/or intestine showed significant differences across the time-points after feeding, but Ghrelin showed no significant difference between T and NT fish in all tested tissues.
Aquaculture Research, 1999
In salmonids, growth hormone (GH) stimulates growth, appetite and the ability to compete for food. This study tested the hypothesis that increased GH levels in GH-transgenic coho salmon Oncorhynchus kisutch (Walbaum) increase competitive ability through higher feeding motivation. The transgenic strain of salmon used contained a gene construct consisting of the sockeye metallothionein-B promoter fused to the type 1 growth gene coding region. The transgenic animals (mean size = 250 g) were F 1 individuals. In six consecutive feeding trials, the intake of contested food pellets by size-matched pairs consisting of one control (1 year older nontransgenic coho salmon) and one GH-transgenic coho salmon was compared. Pellets were provided sequentially until neither ®sh took three consecutive pellets; the identity of the ®sh taking each pellet was noted. Calculated on the three ®rst pellets offered at each feeding trial, the transgenic coho salmon consumed 2.5 times more contested pellets than the controls, supporting the hypothesis that GH transgenesis increases the ability to compete for food. Overall, the transgenic ®sh consumed 2.9 times more pellets that the non-transgenic controls, indicating a high feeding motivation of the transgenic ®sh throughout the feeding trials. It appears that GH transgenesis and GH treatments can induce similar changes in the feeding behaviour of salmonids. Depending on how transgenic and wild individuals differ in other ®tness-related characters, escaped GH transgenic ®sh may compete successfully with native ®sh in the wild.
Proceedings of The National Academy of Sciences, 2009
Domestication has been extensively used in agricultural animals to modify phenotypes such as growth rate. More recently, transgenesis of growth factor genes [primarily growth hormone (GH)] has also been explored as a rapid approach to accelerating performance of agricultural species. Growth rates of many fishes respond dramatically to GH gene transgenesis, whereas genetic engineering of domestic mammalian livestock has resulted in relatively modest gains. The most dramatic effects of GH transgenesis in fish have been seen in relatively wild strains that have undergone little or no selection for enhanced growth, whereas genetic modification of livestock necessarily has been performed in highly domesticated strains that already possess very rapid growth. Such fast-growing domesticates may be refractory to further stimulation if the same regulatory pathways are being exploited by both genetic approaches. By directly comparing gene expression in wild-type, domestic, and GH transgenic strains of coho salmon, we have found that domestication and GH transgenesis are modifying similar genetic pathways. Genes in many different physiological pathways show modified expression in domestic and GH transgenic strains relative to wild-type, but effects are strongly correlated. Genes specifically involved in growth regulation (IGF1, GHR, IGF-II, THR) are also concordantly regulated in domestic and transgenic fish, and both strains show elevated levels of circulating IGF1. Muscle expression of GH in nontransgenic strains was found to be elevated in domesticated fish relative to wild type, providing a possible mechanism for growth enhancement. These data have implications for genetic improvement of existing domesticated species and risk assessment and regulation of emerging transgenic strains.
Aquaculture, 2012
Several different transgenic growth hormone (GH) gene constructs have been used to obtain accelerated growth in salmonids. However, there have been limited direct comparisons of these constructs in terms of the ability to achieve maximal growth in fish. We examined the effect of promoter type (sockeye salmon metallothionein-B or histone 3) fused to a growth hormone-1 coding region from the same species (OnMTGH1 and OnH3GH1 constructs respectively) on growth and plasma growth hormone (GH) and insulin-like growth factor-I (IGF-I) in multiple strains of GH transgenic coho salmon (Oncorhynchus kisutch). Salmon transgenic for the OnMTGH1 construct had consistently greater overall weight than those containing the OnH3GH1 construct, although both groups possessed greatly accelerated growth over non-transgenic fish. However, there were strong strain effects, where some OnH3GH1 strains had similar weight to OnMTGH1 strains while others did not. Triploidy diminished growth acceleration and decreased condition factors in both a fast growing MT strain and slower growing H3 strain. Plasma GH levels did not correlate to weight in transgenic strains, and all but one transgenic strain had plasma GH levels similar to equal sized non-transgenic fish. In contrast, plasma IGF-I content correlated well to size in transgenic strains. The mechanism by which accelerated growth in transgenic fish is obtained appears to be due in part to an upregulation of GH action through increased circulating IGF-I levels, and promoter-type appears to influence potential for growth.
HAYATI Journal of Biosciences, 2019
The animal model response against genetically modified product may provide food safety information. This study was performed to observe behavior, histopathology and physiological responses of Wistar rat fed on the diet containing growth hormone (GH) transgenic common carp (Cyprinus carpio) meal (Ccm). Thirty rats of three-month-old (BW: 115.67–139.50g) were divided into five treatments (six rats per treatment). The treatments were rats fed on the commercial diet without Ccm (control), a re-pelleted diet containing 15% (NT-15) and 45% non-transgenic Ccm (NT-45), a re-pelleted diet containing 15% (TG-15) and 45% GH transgenic Ccm (TG-45). Rats were kept for three weeks, fed twice a day according to treatment by 30 g/day/rat and water was provided ad-libitum. Rat behavior was observed every day during feeding. Serum glutamic oxaloacetic transaminase (SGOT), serum glutamic pyvuric transaminase (SGPT), urea, and creatinine were measured at initial and the end of the experiment. Histolo...
Growth hormone endocrinology of salmonids. Regulatory mechanisms and hormone dynamics
2001
The focus of this review is on the regulatory mechanisms and the mode of action of GH in salmonids. To stimulate further research, it aims at highlighting areas where numerous important breakthroughs have recently been made, as well as where data are currently lacking. The regulation of GH secretion is under complex hypothalamic control, as well as under negative feedback control by GH and IGF-I. Further, the recently characterized ghrelin is a potent GH secretagogue, and may prove to be a link between feed intake and growth regulation. GH plasma profiles show indications of diurnal changes, but whether salmonids have true pulsatile GH secretion remains to be elucidated. The recent cloning and characterization of the salmon GH receptor (GHR) is a major research break-through which will give new insights into the mechanisms of GH action. It should also stimulate research into circulating GH-binding proteins (GHBPs), as they appear to be a soluble form of the GHR. The salmonid GHR sequences show evolutionary divergence from other fish species, but with a high degree of identity within the salmonid group. Radioreceptorassay studies have found GHR present in all tissues examined, which is in line with the highly pleiotropic action of GH. Data are currently scarce on the plasma dynamics of GH in salmonids, and further studies on GHR and GHBPs dynamics coupled to assessments of GH clearance rates and pathways are needed. The direct versus indirect nature of GH action remains to be clarified, but GH appears to act both locally at the target tissue level to stimulate the autocrine/paracrine action of IGF-I, as well as on the liver to increase plasma IGF-I levels. In addition, GH interacts with other hormones such as cortisol, thyroid hormones, insulin, and reproductive hormones, generating a wide range of physiological effects. GH may act both peripherally and directly at the level of the central nervous system to modify behavior, probably by altering the dopaminergic activity in the brain.
Transfer of growth hormone (GH) transgenes into Arctic charr ( Salvelinus alpinus L.)
Genetic Analysis: Biomolecular Engineering, 1999
Four constructs containing salmonid growth hormone (GH) genes were transferred to Arctic charr (Salvelinus alpinus L.). Cytomegalovirus (CMV) and piscine metallothionein B (OnMT) and histone 3 (OnH3) promoters connected to sockeye salmon growth hormone 1 gene (OnGH1) were used for ectopic expression, and Atlantic salmon growth hormone 2 gene with 5′flanking region (SsGH2) was tested for pituitary-specific expression. Charr carrying the OnGH1 constructs showed a dramatic increase in growth rate. The 10-month old transformed fish were 14-fold heavier than control siblings. The ability of the CMVGH1 construct to promote growth was greater than that obtained in fish with piscine promoters. Analysis of individual growth curves of charr carrying the OnH3GH1 transgene indicated a stable ratio of specific growth rates in transformed and control fish regardless of fish size. No alteration in growth performance was found in fish carrying the SsGH2 transgene. There was evidence that the transformed rainbow trout (Oncorhynchus mykiss) were unable to produce SsGH2 mRNA in their pituitary glands. The presence of the transgene in various tissues was examined in trout to evaluate the reliability of one-tissue sampling.