Growth Hormone Transgenic Salmon Research Articles

Rationed and satiated growth hormone transgenic Coho Salmon (Oncorhynchus kisutch) show tissue specific differences in energy stores.

Growth hormone transgenic coho salmon experience increased growth rates, driven primarily through elevated feed intake and feed conversion. However, neuropeptides that signal appetite stimulation have been shown to exhibit variable responses across fed states, suggesting a more complex system mediating growth in these fish. Studies have proposed that growth hormone may have a modulatory role on the energy reserves of fish, possibly through AMP-activated protein kinase (AMPK) activation. AMPK, an energy sensor in cells, has previously been shown to be upregulated in growth hormone transgenic salmon when compared to wild type, however, whether this effect is seen across fed states is unknown. Here, we tested the hypothesis that growth hormone induces an energetic deficit in metabolic tissues, leading to constitutive AMPK activation in growth hormone transgenic salmon. This study compared AMPK activity, ATP, and glycogen, of the liver, heart, and muscle of wild-type, and growth hormone transgenic salmon either fed to satiation or a wild-type ration. The results suggest that white muscle ATP levels in growth hormone salmon are elevated in satiation and rationed conditions. In the liver, growth hormone transgenic salmon fed a rationed wild-type diet experience reductions in ATP level and glycogen. In none of the tissues examined, did AMPK activity change. Taken together, these results indicate that growth hormone transgenic salmon experience metabolic duress when not fed to satiation.

Assessing wild genetic background and parental effects on size of growth hormone transgenic coho salmon

Experiments examining potential impacts of growth hormone (GH) transgenesis in fish typically use a single source strain and do not address potential differential impacts in strains of different genetic backgrounds. Here, we examine the effects of differing genetic backgrounds when reared in culture on the growth of transgenic and nontransgenic coho salmon (Oncorhynchus kisutch) produced by mating sires from different rivers with transgenic dams from a single origin. We found a significant difference in size between offspring of sires originating from various river systems in British Columbia. This difference was independent of differences between transgenotypes (i.e., transgenic vs. nontransgenic offspring). However, the effects of strain or sire were relatively small compared to the effects of the transgene, which were consistent regardless of sire origin. Thus, results derived from studies of GH transgenic fish from a single source population can provide useful information for assessments of GH transgenic salmon from other systems. This has important implications for examining potential risks from introgression of a transgene into different populations.

Comparison of growth rates between growth hormone transgenic and selectively-bred domesticated strains of coho salmon (Oncorhynchus kisutch) assessed under different culture conditions

Aquaculture continues to be the fastest-growing animal food sector and accounts for over 50% of the world's fish consumption. This trend is projected to continue, highlighting the role that the aquaculture industry will play in ensuring future global food security. Enhancement of growth rates of fish can optimize aquaculture production and has been achieved through selective breeding programs producing fast-growing domesticated strains, or the development of growth hormone (GH) transgenic strains. Although the ability of selectively-bred domesticated and GH transgenic salmon to outgrow wild-type salmon is well documented, very little research has been reported that directly compares the growth rates of domesticated and GH transgenic strains. The aim of the present study was to compare the growth and survival of selectively-bred domesticated and GH transgenic coho salmon across a range of experimental conditions. We performed four experiments investigating the growth rates of domesticated and GH transgenic salmon by comparing these strains at different developmental stages and under different experimental rearing conditions, as well as under conditions when strains were competing and when they were reared separately. Results from all experiments showed that GH transgenic coho salmon grew faster and to a larger size than the selectively-bred domesticated coho salmon. This was true whether fish groups were reared post-smolt in freshwater or seawater, whether reared together or separately, and whether manipulated to be size-matched at early stages. Thus, GH transgenesis could become a viable complement to selective breeding to produce fast-growing salmon for aquaculture systems. While further research can continue to optimize the development and use of transgenic strains, the data presented here also are important for environmental risk assessments by providing data on the relative fitness and ecological consequences of selectively-bred domesticated and GH transgenic fish should they enter nature.

Fitness component assessments of wild-type and growth hormone transgenic coho salmon reared in seawater mesocosms

Growth hormone (GH) transgenic fish have been proposed for use in aquaculture to enhance production efficiency. As part of a risk analysis for use of such fish, the influence of GH transgenesis on the potential to persist and succeed in natural ecosystems is being examined in confined laboratory conditions. GH transgenesis can greatly accelerate growth and, in culture conditions, is associated with secondary effects such as poor swimming capacity and spawning success. However, standard culture has also been shown to negatively affect fitness components of wild-type fish, raising the question of whether culture conditions influence fitness components of transgenic fish in a similar way. To examine factors influencing the phenotype of marine-stage GH transgenic salmon (T), and to determine if genotype-by-environment interactions exist at this life stage, we grew T and wild-type (NT) coho salmon (Oncorhynchus kisutch) over six cohort years in 350,000L seawater tanks (termed mesocosms) designed to minimize effects of standard culture conditions. Mesocosm rearing partially facilitated development of normal size and morphology of NT fish relative to nature-reared counterparts, but altered overall body shape, indicating mesocosm conditions do not fully mimic natural environmental effects on coho salmon phenotype. T fish reared in mesocosms had larger mass at maturity than mesocosm- or nature-reared NT fish, indicating GH transgenesis can alter maximum obtainable mass in salmon. Unlike NT, T fish obtained maximum size at maturity across environments, suggesting marine environmental conditions may affect T growth less than NT growth. Screening parents for a common disease agent (Renibacterium salmoninarum) improved seawater survival, and T fish had lower survival than NT fish when from unscreened parents and inconsistent relative survival when from screened parents, indicating GH transgenesis may constitute an advantage or disadvantage in terms of survival. Transgenic salmon had lower swimming capacity and aerobic scope, but similar routine metabolic rate and thermal tolerance, demonstrating transgenesis can have different influences depending on what phenotype is examined. Using an alternate strain of T fish in phenotypic comparisons did not greatly influence most fitness components, although had a strong effect on female fecundity. The inconsistent influence of GH transgenesis on different fitness components, and existence of genotype-by-environment interactions during the marine life stage, complicates extrapolation of laboratory data for transgenic fish to natural environments. However, current and previous data do not provide evidence that overall increased performance of GH transgenic salmon over wild-type fish would arise in the marine environment. Statement of relevanceRearing in seawater mesocosms demonstrate that growth hormone transgenesis has inconsistent effects on marine fitness components in coho salmon.

Rearing in seawater mesocosms improves the spawning performance of growth hormone transgenic and wild-type coho salmon.

Growth hormone (GH) transgenes can significantly accelerate growth rates in fish and cause associated alterations to their physiology and behaviour. Concern exists regarding potential environmental risks of GH transgenic fish, should they enter natural ecosystems. In particular, whether they can reproduce and generate viable offspring under natural conditions is poorly understood. In previous studies, GH transgenic salmon grown under contained culture conditions had lower spawning behaviour and reproductive success relative to wild-type fish reared in nature. However, wild-type salmon cultured in equal conditions also had limited reproductive success. As such, whether decreased reproductive success of GH transgenic salmon is due to the action of the transgene or to secondary effects of culture (or a combination) has not been fully ascertained. Hence, salmon were reared in large (350,000 L), semi-natural, seawater tanks (termed mesocosms) designed to minimize effects of standard laboratory culture conditions, and the reproductive success of wild-type and GH transgenic coho salmon from mesocosms were compared with that of wild-type fish from nature. Mesocosm rearing partially restored spawning behaviour and success of wild-type fish relative to culture rearing, but remained lower overall than those reared in nature. GH transgenic salmon reared in the mesocosm had similar spawning behaviour and success as wild-type fish reared in the mesocosm when in full competition and without competition, but had lower success in male-only competition experiments. There was evidence of genotype×environmental interactions on spawning success, so that spawning success of transgenic fish, should they escape to natural systems in early life, cannot be predicted with low uncertainty. Under the present conditions, we found no evidence to support enhanced mating capabilities of GH transgenic coho salmon compared to wild-type salmon. However, it is clear that GH transgenic salmon are capable of successful spawning, and can reproduce with wild-type fish from natural systems.

Cultured growth hormone transgenic salmon are reproductively out-competed by wild-reared salmon in semi-natural mating arenas

The production of growth hormone (GH) transgenic animals has raised a host of social and scientific concerns regarding their potential impacts on ecosystems should they escape into nature. Indeed, theoretical models suggest that GH transgenic animals could, under specific conditions, decimate local populations. However, while there are numerous laboratory examinations of factors affecting survival of transgenic and wild animals, we know little about the competitive reproductive capacity of transgenic animals. Here, we examined the reproductive capabilities of cultured GH transgenic coho salmon ( Oncorhynchus kisutch) when in competition with wild coho derived from nature using semi-natural mating arenas (within a contained facility). To account for the well-known reproductive impairments associated with culturing salmon in laboratory facilities we contrasted the competitive reproductive success of GH transgenic coho against that of cultured non-transgenic coho. We also performed in vitro sperm analyses to assess the postcopulatory competitive ability of GH transgenic coho. In competitive mating arenas, transgenic coho performed fewer courtship and aggressive behaviours than coho from nature and sired less than 6% of offspring. Non-transgenic cultured coho, despite their smaller body size, sired more than twice as many offspring than transgenic coho when competing against wild coho in mating arenas. Transgenic males also face a postcopulatory reproductive disadvantage as their ejaculates contained fewer sperm that swam slower and for shorter durations than sperm from wild males. Together, these findings suggest limited potential for the transmission of transgenes from cultured GH transgenic coho salmon through natural matings should they escape from a contained culture facility into nature and reproductively interact with a local wild coho salmon strain. However, as responses of wild-reared fish can differ greatly from those of cultured fish, we stress the importance of understanding genotype-by-environment interactions for reproductive phenotypes when developing risk assessment information.

Standing genetic variation and compensatory evolution in transgenic organisms: a growth-enhanced salmon simulation

Genetically modified strains usually are generated within defined genetic backgrounds to minimize variation for the engineered characteristic in order to facilitate basic research investigations or for commercial application. However, interactions between transgenes and genetic background have been documented in both model and commercial agricultural species, indicating that allelic variation at transgene-modifying loci are not uncommon in genomes. Engineered organisms that have the potential to allow entry of transgenes into natural populations may cause changes to ecosystems via the interaction of their specific phenotypes with ecosystem components and services. A transgene introgressing through natural populations is likely to encounter a range of natural genetic variation (among individuals or sub-populations) that could result in changes in phenotype, concomitant with effects on fitness and ecosystem consequences that differ from that seen in the progenitor transgenic strain. In the present study, using a growth hormone transgenic salmon example, we have modeled selection of modifier loci (single and multiple) in the presence of a transgene and have found that accounting for genetic background can significantly affect the persistence of transgenes in populations, potentially reducing or reversing a “Trojan gene” effect. Influences from altered life history characteristics (e.g., developmental timing, age of maturation) and compensatory demographic/ecosystem controls (e.g., density dependence) also were found to have a strong influence on transgene effects. Further, with the presence of a transgene in a population, genetic backgrounds were found to shift in non-transgenic individuals as well, an effect expected to direct phenotypes away from naturally selected optima. The present model has revealed the importance of understanding effects of selection for background genetics on the evolution of phenotypes in populations harbouring transgenes.

Endocrine effects of growth hormone overexpression in transgenic coho salmon

Non-transgenic (wild-type) coho salmon ( Oncorhynchus kisutch), growth hormone (GH) transgenic salmon (with highly elevated growth rates), and GH transgenic salmon pair fed a non-transgenic ration level (and thus growing at the non-transgenic rate) were examined for plasma hormone concentrations, and liver, muscle, hypothalamus, telencephalon, and pituitary mRNA levels. GH transgenic salmon exhibited increased plasma GH levels, and enhanced liver, muscle and hypothalamic GH mRNA levels. Insulin-like growth factor-I (IGF-I) in plasma, and growth hormone receptor (GHR) and IGF-I mRNA levels in liver and muscle, were higher in fully fed transgenic than non-transgenic fish. GHR mRNA levels in transgenic fish were unaffected by ration-restriction, whereas plasma GH was increased and plasma IGF-I and liver IGF-I mRNA were decreased to wild-type levels. These data reveal that strong nutritional modulation of IGF-I production remains even in the presence of constitutive ectopic GH expression in these transgenic fish. Liver GHR membrane protein levels were not different from controls, whereas, in muscle, GHR levels were elevated approximately 5-fold in transgenic fish. Paracrine stimulation of IGF-I by ectopic GH production in non-pituitary tissues is suggested by increased basal cartilage sulphation observed in the transgenic salmon. Levels of mRNA for growth hormone-releasing hormone (GHRH) and cholecystokinin (CCK) did not differ between groups. Despite its role in appetite stimulation, neuropeptide Y (NPY) mRNA was not found to be elevated in transgenic groups.

Tissue specific expression of antifreeze protein and growth hormone transgenes driven by the ocean pout (Macrozoarces americanus) antifreeze protein OP5a gene promoter in Atlantic salmon (Salmo salar)

Previous research aimed at producing genetically improved salmon broodstock for aquaculture led to the creation of two lines of transgenic Atlantic salmon using gene constructs that were derived in part from the ocean pout OP5a antifreeze protein (AFP) gene. One of the lines was produced using an OP5a AFP gene in which the 5' region of the promoter was removed (termed t-OP5a-AFP), and the other line contains a growth hormone (GH) transgene (EO-1alpha) that consists of a chinook salmon GH cDNA driven by a truncated OP5a AFP promoter that is almost identical to that of the t-OP5a-AFP construct. The similarity of the promoter regions of these transgenes provided an opportunity to evaluate their tissue specific expression patterns. Expression of mRNA was evaluated using Northern blot and RT-PCR techniques. The results demonstrate that the AFP and GH trangenes were expressed in almost all body tissues, suggesting that the promoter region of the OP5a AFP gene lacks tissue specific elements. Northern analysis revealed that expression of the t-OP5a-AFP gene was considerably greater than that of the EO-1alpha GH transgene. Only the spleen tissue of the GH transgenics showed a visible band of hybridization. In contrast clear bands of hybridization were evident in all tissues, except for blood cells, of the AFP transgenics with heart, liver and brain tissue showing the highest levels of mRNA expression. This higher level of expression could be attributable to the presence of introns in the t-OP5a-AFP transgene. Since the GH transgenic salmon grow considerably faster than non-transgenics the low levels of GH transgene expression in this line were clearly sufficient to produce the desired rapid growth phenotype. In contrast the levels of AFP expression were inadequate to impart any improvement in the freeze resistance of the AFP transgenic salmon.

The glutathione antioxidant system is enhanced in growth hormone transgenic coho salmon (Oncorhynchus kisutch)

Insertion of a growth hormone (GH) transgene in coho salmon results in accelerated growth, and increased feeding and metabolic rates. Whether other physiological systems within the fish are adjusted to this accelerated growth has not been well explored. We examined the effects of a GH transgene and feeding level on the antioxidant glutathione and its associated enzymes in various tissues of coho salmon. When transgenic and control salmon were fed to satiation, transgenic fish had increased tissue glutathione, increased hepatic glutathione reductase activity, decreased hepatic activity of the glutathione synthesis enzyme gamma-glutamylcysteine synthetase, and increased intestinal activity of the glutathione catabolic enzyme gamma-glutamyltranspeptidase. However, these differences were mostly abolished by ration restriction and fasting, indicating that upregulation of the glutathione antioxidant system was due to accelerated growth, and not to intrinsic effects of the transgene. Increased food intake and ability to digest potential dietary glutathione, and not increased activity of glutathione synthesis enzymes, likely contributed to the higher levels of glutathione in transgenic fish. Components of the glutathione antioxidant system are likely upregulated to combat potentially higher reactive oxygen species production from increased metabolic rates in GH transgenic salmon.

Cardiorespiratory modifications, and limitations, in post-smolt growth hormone transgenic Atlantic salmonSalmo salar

In recent years, there has been a great deal of interest in how growth hormone (GH) transgenesis affects fish physiology. However, the results of these studies are often difficult to interpret because the transgenic and non-transgenic fish had very different environmental/rearing histories. This study used a stable line of size-matched GH Atlantic salmon (Salmo salar) that were reared in a shared tank with controls (at 10 degrees C, for approximately 9 months) to perform a comprehensive examination of the cardiorespiratory physiology of GH transgenic salmon, and serves as a novel test of the theory of symmorphosis. The GH transgenic salmon had a 3.6x faster growth rate, and 21 and 25% higher values for mass-specific routine and standard oxygen consumption (M(O(2))), respectively. However, there was no concurrent increase in their maximum M(O(2)), which resulted in them having an 18% lower metabolic scope and a 9% reduction in critical swimming speed. This decreased metabolic capacity/performance was surprising given that the transgenics had a 29% larger heart with an 18% greater mass-specific maximum in situ cardiac output, a 14% greater post-stress blood haemoglobin concentration, 5-10% higher red muscle and heart aerobic enzyme (citrate synthase or cytochrome oxidase) activities, and twofold higher resting and 1.7x higher post-stress, catecholamine levels. However, gill surface area was the only cardiorespiratory parameter that was not enhanced, and our data suggest that gill oxygen transfer may have been limiting. Overall, this research: (1) shows that there are significant metabolic costs associated with GH transgenesis in this line of Atlantic salmon; (2) provides the first direct evidence that cardiac function is enhanced by GH transgenesis; (3) shows that a universal upregulation of post-smolt (adult) GH transgenic salmon cardiorespiratory physiology, as suggested by symmorphosis, does not occur; and (4) supports the idea that whereas differences in arterial oxygen transport (i.e. cardiac output and blood oxygen carrying capacity) are important determinants of inter-specific differences in aerobicity, diffusion-limited processes must be enhanced to achieve substantial intra-specific improvements in metabolic and swimming performance.

Genes Differentially Expressed in Growth Hormone Transgenic Salmon.

Previous articleNext article No AccessProtein Chemistry, Molecular Biology, and EvolutionGenes Differentially Expressed in Growth Hormone Transgenic SalmonKristen M. Ettensohn, Peggy Biga, Christina Romano, Robert H. Devlin, and Steven B. RobertsKristen M. Ettensohn1. Marine Biological Laboratory, Woods Hole, Massachusetts2. Dartmouth College, Hanover, New Hampshire Search for more articles by this author , Peggy Biga3. Great Lakes WATER Institute, Milwaukee, Wisconsin Search for more articles by this author , Christina Romano1. Marine Biological Laboratory, Woods Hole, Massachusetts Search for more articles by this author , Robert H. Devlin4. Fisheries and Oceans Canada, West Vancouver, British Columbia, Canada Search for more articles by this author , and Steven B. Roberts1. Marine Biological Laboratory, Woods Hole, Massachusetts Search for more articles by this author PDFPDF PLUSFull Text Add to favoritesDownload CitationTrack CitationsPermissionsReprints Share onFacebookTwitterLinkedInRedditEmail SectionsMoreDetailsFiguresReferencesCited by The Biological Bulletin Volume 207, Number 2October 2004 Published in association with the Marine Biological Laboratory Article DOIhttps://doi.org/10.1086/BBLv207n2p168a Views: 33Total views on this site PDF download Crossref reports no articles citing this article.

Growth hormone transgenic salmon pay for growth potential with increased predation mortality

Recent advances in gene technology have been applied to create fast-growing transgenic fish, which are of great commercial interest owing to their potential to shorten production cycles and increase food production. However, there is growing concern and speculation over the impact that escaped growth hormone (GH)-transgenic fish may have on the natural environment. To predict these risks it is crucial to obtain empirical data on the relative fitness of transgenic and non-transgenic fish under nature-like conditions. Using landscaped stream aquaria with live food and predators, we show that the predation mortality of newly hatched GH-transgenic coho salmon fry (Oncorhynchus kisutch) is much higher than in non-transgenic conspecifics, and that this difference is amplified when food abundance decreases. The growth rate of transgenic and non-transgenic fish is similar at high food levels, whereas transgenic fish grow more slowly than non-transgenic fish when food abundance is reduced. Our results suggest that the fitness of young GH-transgenic coho salmon in the wild will be determined by both predation pressure and food availability.

Growth, viability and genetic characteristics of GH transgenic coho salmon strains

Transgenic coho salmon strains containing an all-salmon growth hormone (GH) gene construct (OnMTGH1) have been examined. The transgene utilized is comprised of the metallothionein-B promoter driving the expression of the type-I growth hormone gene from sockeye salmon. Transgene DNA is integrated into the salmon genome, but is transmitted at low frequency from founder transgenic animals consistent with late integration following microinjection resulting in mosaic animals, whereas subsequent generations transmit transgene DNA as a stable Mendelian trait. Different families established from separate founder animals yield lines with unique growth characteristics suggesting important site-of-integration effects on transgene expression. Growth enhancement of transgenic salmon is initiated early, with advanced hatch timing but occurring also throughout the life history, particularly during the early phase in fresh water. GH transgenic fish showed precocious smoltification and onset of sexual maturation, but approximately normal adult body size, indicating that compression of the normal coho salmon life history is occurring. The viability of diploid GH transgenic salmon ranges from reductions to greater than that of controls among strains, and triploid transgenic animals had normal viability relative to diploid transgenic salmon. Triploid transgenic salmon display a reduction in growth rate relative to transgenic diploids, but are still significantly growth enhanced compared with nontransgenic controls. The distinct phenotypic characteristics of GH transgenic families suggest that evaluation for aquaculture and for risk assessments requires examination of strains on a case-by-case basis. Furthermore, strong effects of size at maturity in culture conditions were observed for nontransgenic wild strain coho salmon which were not apparent in GH transgenic salmon, indicating that strong genotype by environment effects strongly influence phenotype and complicate risk assessments.

Transgene and host growth hormone gene expression in pituitary and nonpituitary tissues of normal and growth hormone transgenic salmon

Growth hormone (GH) gene expression has been examined in control and transgenic coho salmon containing a transgene comprised of the sockeye salmon GH1 gene under the control of the MT-B promoter from the same species. This transgene dramatically enhances the growth of salmonids, and raises serum GH levels some forty-fold. Transcript levels from this transgene were detected by RT-PCR using construct-specific GH primers in all tissues examined (liver, kidney, skin, intestine, stomach, muscle, spleen, pyloric caeca), and ranged from 0.1∼9.4 pg/50 μg total RNA in different tissues as estimated by dot blot analysis. Interestingly, GH gene expression was also observed in intestine of control coho salmon by RT-PCR capable of detecting host and transgene transcripts using general primers. Sequence analysis of the intestinal GH mRNA from controls indicated it was derived from the coho GH2 gene. GH mRNA abundance analyzed by northern analysis indicates lower levels are found in large (400–500 g) than small transgenic salmon (20–21 g). No molecular evidence for transgene expression was obtained in tissues from transgenic fry, despite an obvious increase in size relative to control siblings, suggesting very low levels of transgene expression early in development. GH mRNA levels (per μg RNA) were also examined in the pituitary gland, and were found to be significantly lower ( P<0.01) in transgenic coho compared to nontransgenic animals of the same size. Pituitary glands of transgenic animals were also smaller than control animals of the same size, and pituitary size, expressed as a proportion of body weight, decreased with body size in transgenic but not control animals. These results imply that pituitary GH expression is regulated by negative feed-back controls as occurs in other vertebrate systems. GH mRNA was examined in pituitary glands by whole-mount in situ hybridization, and, whereas overall levels appeared reduced in transgenic animals, the site of hybridization did not differ between transgenic and control glands.