Trout Muscle Research Articles

Genomic structure and expression of uncoupling protein 2 genes in rainbow trout (Oncorhynchus mykiss)

BackgroundUncoupling protein 2 (UCP2) belongs to the superfamily of mitochondrial anion carriers that dissociate the respiratory chain from ATP synthesis. It has been determined that UCP2 plays a role in several physiological processes such as energy expenditure, body weight control and fatty acid metabolism in several vertebrate species. We report the first characterization of UCP2s in rainbow trout (Oncorhynchus mykiss).ResultsTwo UCP2 genes were identified in the rainbow trout genome, UCP2A and UCP2B. These genes are 93% similar in their predicted amino acid sequences and display the same genomic structure as other vertebrates (8 exons and 7 introns) spanning 4.2 kb and 3.2 kb, respectively. UCP2A and UCP2B were widely expressed in all tissues of the study with a predominant level in macrophage-rich tissues and reproductive organs. In fry muscle we observed an increase in UCP2B expression in response to fasting and a decrease after refeeding in agreement with previous studies in human, mouse, rat, and marsupials. The converse expression pattern was observed for UCP2A mRNA which decreased during fasting, suggesting different metabolic roles for UCP2A and UCP2B in rainbow trout muscle. Phylogenetic analysis including other genes from the UCP core family located rainbow trout UCP2A and UCP2B with their orthologs and suggested an early divergence of vertebrate UCPs from a common ancestor gene.ConclusionWe characterized two UCP2 genes in rainbow trout with similar genomic structures, amino acid sequences and distribution profiles. These genes appeared to be differentially regulated in response to fasting and refeeding in fry muscle. The genomic organization and phylogeny analysis support the hypothesis of a common ancestry between the vertebrate UCPs.

Microarray gene expression analysis in atrophying rainbow trout muscle: a unique nonmammalian muscle degradation model

Muscle atrophy is a physiological response to diverse physiological and pathological conditions that trigger muscle deterioration through specific cellular mechanisms. Despite different signals, the biochemical changes in atrophying muscle share many common cascades. Muscle deterioration as a physiological response to the energetic demands of fish vitellogenesis represents a unique model for studying the mechanisms of muscle degradation in non-mammalian animals. A salmonid microarray, containing 16,006 cDNAs, was used to study the transcriptome response to atrophy of fast-switch muscles from gravid rainbow trout compared with sterile fish. Eighty-two unique transcripts were upregulated and 120 transcripts were downregulated in atrophying muscles. Transcripts having gene ontology identifiers were grouped according to their functions. Muscle deterioration was associated with elevated expression of genes involved in the catheptic and collagenase proteolytic pathways; the aerobic production, buffering, and utilization of ATP; and growth arrest; whereas atrophying muscle showed downregulation of genes encoding a serine proteinase inhibitor, enzymes of anaerobic respiration, muscle proteins as well as genes required for RNA and protein biosynthesis/processing. Therefore, gene transcription of the trout muscle atrophy changed in a manner similar to mammalian muscle atrophy. These changes result in an arrest of normal cell growth, protein degradation, and decreased glycolytic cellular respiration that is characteristic of the fast-switch muscle. For the first time, other changes/mechanisms unique to fish were discussed including genes associated with muscle atrophy.

Steroid-induced cardiac contractility requires exogenous glucose,glycolysis and the sarcoplasmic reticulum in rainbow trout

Recent data from our laboratory suggest that sex steroids promote contractile function in cardiac muscle of rainbow trout (Oncorhynchus mykiss Walbaum), and there are sex differences in hormone signaling and cardiac function. The current study investigated whether steroid-induced inotropism in electrically paced (0.5 Hz, 14 degrees C) ventricle strips at 90% Lmax (1) has a metabolic requirement for exogenous glucose and (2) is associated with enhanced intracellular Ca2+ storage and release from the sarcoplasmic reticulum (SR). We also explored whether sex differences exist in extracellular Ca2+ (Ca2+o) or cardiac sensitivity to Ca2+o. In the absence or at low concentrations (1 or 2 mmol l-)) of exogenous glucose, resting tension and relaxation time were increased selectively in cardiac tissue from females. Increasing glucose promoted twitch force in a bell-shaped manner, with 5 mmol l-1 representing the optimal concentration for both sexes. The positive inotropic effects of physiological concentrations of testosterone (T) and 17beta-estradiol (E2) in male and female trout ventricle strips, respectively, developed slowly (10-45 min) and were not apparent in glucose-free medium, in medium containing iodoacetate (IAA), an inhibitor of glycolysis, or medium containing 5 mmol l-) lactate or pyruvate. Male ventricle strips had increased inotropic responses to glucose and T compared with female strips exposed to glucose and E2. Furthermore, sexually maturing males showed a greater inotropic response than immature males or females. Pretreatment with ryanodine (a specific blocker of SR Ca2+ release) also eliminated the inotropic effects of sex steroids and exogenous glucose and reduced the post-rest potentiation of contractile force (a marker of SR Ca2+ storage). By contrast, the inotropic effects of epinephrine (Epi) or elevated Ca2+o were faster (developing within 1-3 min) and were not diminished by the presence or absence of glucose or by pretreatment with IAA or ryanodine. Sex differences were also found in responsiveness to caffeine (males>females) and the relationship between Ca2+ concentration and force development above baseline. The Ca2+50 was lower in female cardiac tissue than males, suggesting greater Ca2+ sensitivity, and although plasma albumin was higher in females, total and ionized plasma Ca2+ did not differ between the sexes. For the first time, our study highlights the importance of extracellular glucose, glycolytic activity and SR Ca2+ storage and release for sex steroid-induced inotropism in the trout ventricle. Conversely, the inotropes Epi and elevated [Ca2+o] do not require the presence or metabolism of exogenous glucose or the SR for signaling their positive effects on contractility. These results also demonstrate novel sex-related differences in cardiac reliance on exogenous glucose, Ca2+ sensitivity and SR function and thus should be considered in future studies.

Effect of Varying the Concentration of Dietary Astaxanthin on Rainbow Trout, Oncorhynchus mykissMuscle Pigmentation

The effect of daily distribution of various dietary astaxanthin concentrations (0, 10, 15, 25, and 30 mg astaxanthin/kg diet) on rainbow trout muscle pigmentation was studied during 9 weeks. Four combined daily distribution treatments (morning/evening) of dietary astaxanthin (25/25, 25/10, 15/15, and 30/0) were tested on triplicate groups of 80 immature rainbow trout (body weight±SD, 199.6±3.7 g). Trout muscle pigmentation was evaluated on the basis of muscle astaxanthin retention and muscle color. Fish muscle astaxanthin increased with increasing dietary carotenoid concentration. There was no significant (P > 0.05) difference for muscle astaxanthin retention coefficients and muscle color parameters of fish receiving treatments 25/25 and 25/ 10 while there was a significant (P < 0.05) difference for muscle astaxanthin retention coefficients and muscle color parameters of fish receiving treatments 25/25 and 15/15 or 30/0. Astaxanthin retention coefficients were higher for lower dietary doses (15/15 or 30/0) while muscle color parameters were lower for fish receiving treatments 15/15 or 30/0. These results suggest that the level of dietary astaxanthin currently used for trout pigmentation (25/25) may be reduced by 30% by varying its daily distribution in feeds (25/10).

Fasting and refeeding affect the expression of the Inhibitor of DNA Binding (ID) genes in rainbow trout ( Oncorhynchus mykiss) muscle

The Inhibitor of DNA Binding/Differentiation (ID) proteins are a family of dominant negative regulators of the basic helix-loop-helix (bHLH) transcription factors, shown in mammals to delay cell differentiation and prolong proliferation. In the current study we used real-time PCR to investigate the effects of fasting and refeeding on the expression of ID genes in rainbow trout muscle. Fry shortly following yolk-sac absorption (∼ 250 mg) were used in a pair of experiments. In the first experiment, the treatment groups included fish fed or fasted throughout the duration of the experiment, and fish fasted for 14 days followed by feeding for the remainder of the experiment. The second experiment consisted of the same treatment groups; however the fish were only fasted for 7 days prior to refeeding. In both experiments, ID gene expression in the muscle of fasted fish was significantly lower than the fed samples after 7 days. Refeeding for 3 or 7 days returned the ID expression to levels similar to the fed fish. The reduction of ID expression during a fast and the subsequent return to fed levels with refeeding suggests the ID proteins participate in the regulation of muscle growth in the rainbow trout.

Comparison of red chilli ( Capsicum annuum) oleoresin and astaxanthin on rainbow trout ( Oncorhyncus mykiss) fillet pigmentation

The effect of dietary carotenoid from red chilli oleoresin and synthetic astaxanthin on the pigmentation of fresh and minced fillet of rainbow trout was compared. Five experimental diets were evaluated: a control diet (CD) containing 80 mg synthetic astaxanthin (Carophyll Pink 8%) kg − 1 diet, two diets containing 80 mg carotenoid kg − 1 diet (HRL 1, HRL 2), and two diets containing 120 mg carotenoid kg − 1 diet (HRH 1, HRH 2), each paired with low and high carotenoid containing diets from red chilli oleoresins containing different carotenoid composition and concentration, respectively. The six-week experiment was carried out in a closed recirculation system consisting of ten 3.0 m 3 circular tanks containing 161 (150 ± 14 g) rainbow trout. The trout were fed daily at 3% of their total biomass. Results indicated that significant differences existed between fresh and minced fillet colour. Diet HRH 2 showed the least colour difference between the fresh and minced fillets, suggesting that the incorporation of 120 mg carotenoid from red chilli oleoresin kg − 1 diet allowed for processing of fillets without significant colour loss. All of the diets produced carotenoid concentrations in trout muscle of more than 5 mg carotenoid kg − 1 muscle at the end of the experimental trial. Colour parameters and carotenoid concentrations in muscle were linearly related.

Determination of 17α-Methyltestosterone in Muscle Tissues of Tilapia, Rainbow Trout, and Salmon Using Liquid Chromatography−Tandem Mass Spectrometry

An analytical method was developed to quantitate and confirm the presence of 17alpha-methyltestosterone in the muscles of tilapia, rainbow trout, and salmon. The method employed two liquid-liquid partitioning steps and two solid-phase extraction columns for sample cleanup. The final extracts were analyzed on an isocratic reverse-phase liquid chromatography-tandem mass spectrometry system with atmospheric-pressure chemical ionization in the positive ion mode. The method was validated at levels from 0.40 to 1.6 ng/g, with MT-d3 used as an internal standard. The accuracy was between 100% and 110%, and coefficients of variation of <10% were obtained for all three fish species. Muscle tissues from dosed fish were also assayed to demonstrate the effectiveness of the method for recovering the parent drug.

Differences in some properties of lactate dehydrogenase from muscles of the carp Cyprinus carpio and trout Salmo gairdneri

Some catalytic and kinetic properties of lactate dehydrogenase (LDH) isolated from trout and carp skeletal muscles were compared. The specific activity of LDH in the carp muscle was lower by about one third than the activity in the trout muscle. Temperature and pH optima for LDH isolated from the carp muscle were higher than those for the trout muscle LDH. Moreover, in direct reaction, the carp muscle LDH had a higher affinity both for pyruvate and for NADH, i.e., it had lower KM values. Instead, the trout muscle LDH showed the positive kinetic cooperativity (the Hill coefficient > 1) of the substrate and coenzyme binding sites. Thus, the carp LDH seems to function more effectively under anaerobic conditions and at higher temperatures.

Anabolic effects of feeding β 2-adrenergic agonists on rainbow trout muscle proteases and proteins

β 2-Adrenergic agonists (BAAs) act as repartitioning agents in domestic animals by redistributing nutrients away from adipose tissue and towards muscle accretion. The mechanism involves altering the rates of protein degradation and synthesis. The aim of this study was to test the effects of chronic feeding of the BAAs clenbuterol (CLEN) and ractopamine (RACT) on rainbow trout (RBT) muscle. Specifically, we examined the activities and mRNA levels of genes in the major proteolytic pathways including calpains, the multi-catalytic proteasome and cathepsins, and the mRNA levels of genes encoding the myofibrillar proteins, fast-twitch and slow-twitch myosin heavy chains (f-MHC and s-MHC, respectively), and the cytoskeletal protein, β-actin. RACT feeding significantly increased mRNA transcripts of the calpain catalytic subunit (Capn1), the regulatory subunit (cpns), and the calpastatin large isoform (CAST-L), without affecting the calpain enzyme activity. CLEN feeding significantly increased mRNA levels of the proteasome alpha subunit without a corresponding change in 20S enzyme activity. RACT significantly decreased cathepsin D activity without affecting mRNA levels suggesting that the action of RACT may be at the post-transcriptional level. In addition, both CLEN and RACT significantly increased mRNA transcripts of f-MHC and β-actin genes suggesting an anabolic role of BAAs on myofibrillar and cytoskeletal proteins. Neither CLEN nor RACT altered mRNA expression of the s-MHC gene indicating no transformation of muscle fiber-types. This study supports a role for BAAs in inducing RBT muscle accretion by altering both protein synthesis and degradation.

Peptides in rainbow trout (Oncorhynchus mykiss) muscle subjected to ice storage and cooking

Abstract Although proteolysis during post mortem storage is an important factor affecting fish texture, little is known about degradation end-products. This study was performed to investigate the occurrence of low molecular weight peptides ( in post mortem rainbow trout muscle, during ice storage, and to evaluate their stability during cooking. It combined quantitative (amino acid analysis) and qualitative approaches (mass spectrometry). The results showed that muscle of trout was poor in peptides. These were mainly anserine and glutathione. Their concentration was almost unaffected by the seven days of ice storage and vacuum cooking for 5 min at 70 °C. MS analysis revealed a limited but highly reproducible appearance of small peptides in trout muscle during the ice storage and after cooking. The compounds detected by MS analysis remain to be characterised.

Molecular characterization of muscle atrophy and proteolysis associated with spawning in rainbow trout

Severe muscle deterioration is a physiological response to the energetic demands of fish spawning. This response represents a suitable model to study mechanisms of muscle degradation in fish where typical tetrapod methods, such as muscle unloading, are not applicable. Enzyme activities and mRNA accumulations of genes in major proteolytic pathways, including cathepsins, calpains and the multi-catalytic proteasome, were measured in white muscles of rainbow trout during spawning and post-spawning seasons of gravid fish for comparisons to sterile fish. Fertile fish at spawning had less muscle tissue and less muscle protein compared to sterile fish and post-spawning fertile fish. Muscle deterioration of the fertile fish during spawning was associated with greater mRNA accumulation and elevated activity of cathepsin-L. Concurrently, muscle of spawning fish showed increased mRNA accumulations of cathepsin-D, the calpain regulatory subunit and the proteasome catalytic subunit alpha without corresponding increases in enzyme activities. In addition, elevated activity and increased mRNA accumulation of caspase-9, but not caspase-3, were observed in fertile fish during spawning. This study indicates that cathepsins mediate protein catabolism during spawning in rainbow trout and the catabolic process may involve activation of the apoptosis mediator, caspase-9, but not the apoptosis executioner, caspase-3.

IGF-I and insulin receptor signal transduction in trout muscle cells

In this study, primary cultures of trout skeletal muscle cells were used to investigate the main signal transduction pathways of insulin and IGF-I receptors in rainbow trout muscle. At different stages of in vitro development (myoblasts on day 1, myocytes on day 4, and fully developed myotubes on day 11), we detected in these cells the presence of immunoreactivity against ERK 1/2 MAPK and Akt/PKB proteins, components of the MAPK and the phosphatidylinositol 3-kinase-Akt pathways, respectively, two of the main intracellular transduction pathways for insulin and IGF-I receptors. Both insulin and IGF-I activated both pathways, although the latter provoked higher immunoreactivity of phosphorylated MAPKs and Akt proteins. At every stage, increases in total MAPK immunoreactivity levels were observed when cells were stimulated with IGF-I or insulin, while total Akt immunoreactivity levels changed little under stimulation of peptides. Total Akt and total MAPK levels increased as skeletal muscle cells differentiated in culture. Moreover, when cells were incubated with IGF-I or insulin, MAPK-P immunoreactivity levels showed greater increases over the basal levels on days 1 and 4, with no effect observed on day 11. Although Akt-P immunoreactivity displayed improved responses on days 1 and 4 as well, a stimulatory effect was still observed on day 11. In addition, the present study demonstrates that purified trout insulin receptors possess higher phosphorylative activity per unit of receptor than IGF-I receptors. In conclusion, these results indicate that trout skeletal muscle culture is a suitable model to study the insulin and IGF-I signal transduction molecules and that there is a different regulation of MAPK and Akt pathways depending on the developmental stage of the muscle cells.

Trace Elements as Fingerprint of Lake of Provenance and of Species of Some Native and Exotic Fish of Northern Patagonian Lakes

A survey of trace element contents in fish muscle and liver was performed in different lakes of two northern Patagonian national parks: Nahuel Huapi and Los Alerces national parks. The aim of the work was to obtain the first set of reference data on elements that are not liable to be disturbed by human activities and to identify compositional patterns related to the species and site of collection. The species studied are native creole perch and velvet catfish and exotic brown trout, rainbow trout, and brook trout. The elements analyzed are Br, Cs, Fe, Rb, Se, Na, and Zn. Trace elements in muscle of brown trout, rainbow trout, and creole perch showed statistical patterns that allow one to identify the national park of origin, as well as which of the lakes (Traful, Espejo Chico, or the Nahuel Huapi-Moreno system) of the Nahuel Huapi National Park from which they come. Contents in the liver provide similar but less clear patterns than contents in muscle; however, in some particular cases, they provide additional information. Trace element contents in muscle are also good indicators of the species collected within a lake.

Changes in concentrations of microcystins in rainbow trout, freshwater mussels, and cyanobacteria in Lakes Rotoiti and Rotoehu

Microcystin concentrations in cyanobacteria and their accumulation in rainbow trout (Oncorhynchus mykiss) and freshwater mussels (Hyridella menziesi) in Lakes Rotoiti and Rotoehu (New Zealand) were investigated. Hatchery rainbow trout were added to an enclosure in Lake Rotoiti where concentrations of microcystins in the phytoplankton and cyanobacterial cell concentrations could be closely monitored. Rainbow trout that were free to roam in the entire area of each lake were also included in the study. Freshwater mussels were suspended subsurface in cages in the enclosure. Phytoplankton samples, rainbow trout liver and muscle tissue, and the tissues of mussels were analyzed for microcystins using the ADDA-ELISA method, and selected samples were analyzed using LC-MS. A maximum concentration of microcystins in the phytoplankton samples of 760 microg L(-1) was recorded in Te Weta Bay, Lake Rotoiti, in March 2004. ELISA results confirmed microcystin immunoreactivity in rainbow trout liver and muscle tissues and in freshwater mussels. The microcystin congeners LR, YR, RR, AR, FR, LA, and WR were detected by LC-MS in caged freshwater mussels in Lake Rotoiti but were not detected in either muscle or liver tissue of rainbow trout. The daily tolerable intake limit of microcystins for human consumption recommended by the World Health Organisation is 0.04 microg kg(-1) day(-1). Modeling was carried out for the human intake of microcystin compounds from rainbow trout muscle tissue, and the potential health risks were estimated, assuming the ADDA-ELISA was determining compounds of toxicity equivalent to microcystin-LR.

Rapid Near Infrared Spectroscopic Method for the Detection of Spoilage in Rainbow Trout (Oncorhynchus mykiss) Fillet

ABSTRACT:The possibility of using visible and short‐wavelength near‐infrared (SW‐NIR: 600 to 1100 nm) spectroscopy to detect the onset of spoilage and to quantify microbial loads in rainbow trout (Oncorhynchus mykiss) was investigated. Spectra were acquired on the skin and flesh side of intact trout fillet portions and on minced trout muscle samples stored at 4 °C for up to 8 d or at room temperature (21 °C) for 24 h. Principal component analysis (PCA) and partial least squares (PLS) chemometric models were developed to predict the onset and degree of spoilage. PCA results showed clear segregation between the control (day 1) and the samples held 4 d or longer at 4 °C. Clear segregation was observed for samples stored 10 h or longer at 21 °C compared with the control (0 h), indicating that onset of spoilage could be detected with this method. Quantitative PLS prediction models for microbial loads were established. For trout fillets, 4 °C: R= 0.97, standard error of prediction (SEP) = 0.38 log colony‐forming units (CFU)/g (flesh side); R= 0.94, SEP = 0.53 log CFU/g (skin side); R= 0.82, SEP = 0.82 log CFU/g for minced fish held at 21 °C. These results indicate that SW‐NIR in combination with multivariate statistical methods can be used to detect and monitor the spoilage process in rainbow trout and quantify microbial loads rapidly and accurately.

The replacement of fish oil with refined coconut oil in the diet of large rainbow trout (Oncorhynchus mykiss)

Three hundred rainbow trout (242.9 ± 6.9 g) were randomly assigned to 12 fibreglass tanks (25 fish/tank). The fish were fed, for 231 days (feeding level: 0.74 % average bw), four experimental diets, containing increasing levels of coconut oil: diet A 0 %, diet B 6 %; diet C 12 % and diet D 13 %, as replacements of herring oil and cod liver oil. Weight gains of fish were > 3 g d-1 and FCR lower than 1.5, for all treatments. No significant difference was observed in the main carcass traits and whole body composition, but whole fish energy content significantly increased in fish fed diet D (10.68 KJ g-1). Different dietary fatty acid profiles had significant effects on the rainbow trout muscle contents of C10:0, C12:0, C14:0, total n-3, and the main ratios: SFA/USFA; DHA/EPA; DHA/AA, n-3/n-6, but not on total fatty acids content of the muscle.

Effect of carotenoids from red pepper and marigold flower on pigmentation, sensory properties and fatty acid composition of rainbow trout

Effects of carotenoid sources on pigmentation, sensory properties and fatty acid composition of rainbow trout ( Onchorhynchus mykiss) were investigated. The fish (120.51 ± 0.75 g) were fed with diets containing 1.8% marigold flower, 5% red pepper, 70 mg kg −1 commercial astaxanthin and compared with a control group for 60 days. Commercial astaxanthin provided the highest carotenoid accumulation in the fish, and this was followed by red pepper and marigold flower ( p < 0.05). Dietary carotenoid sources did not significantly affect fatty acid composition of the fish fillets. Trout muscle coloured with commercial astaxanthin was more preferred than the others by the sensory panellists.

Thiamine Status of Cayuga Lake Rainbow Trout and Its Influence on Spawning Migration

AbstractRainbow trout Oncorhynchus mykiss in Cayuga Lake, New York, appear to be suffering from a thiamine deficiency because their progeny develop general weakness, loss of equilibrium, and increased mortality, which are prevented by treatment with thiamine. Thiamine status and its effect on adults are unknown. In 2000 and 2002, we captured, tagged, and released 64 and 189 prespawning rainbow trout, respectively, in Cayuga Inlet at a collection weir to evaluate their thiamine status and the effect of thiamine injection (150 nmol/g) on instream migration. Half of the rainbow trout in each year (32 in 2000 and 95 in 2002) were injected with thiamine and half were uninjected; all rainbow trout were released above the weir to continue their upstream migration. By means of electrofishing in 2000, we recaptured significantly more thiamine‐injected (N = 7) than uninjected (N = 0) rainbow trout approximately 7.0–9.3 river kilometers upstream from the weir. In 2002, the concentration of thiamine in the muscle of rainbow trout collected above a 1.8‐m cascade was significantly higher (mean ± SD = 5.47 ± 5.04 nmol/g; range = 1.0– 13.8 nmol/g; N = 8) than that of rainbow trout collected either above a 1.0‐m cascade (1.36 ± 0.71 nmol/g; range = 0.6–3.3 nmol/g; N = 16) or below the cascades (1.20 ± 0.46 nmol/g; range = 0.7–1.9 nmol/g; N = 5). The lowest concentration of thiamine observed in the muscle of rainbow trout collected upstream of the 1.8‐m cascade was 1.0 nmol/g, suggesting that the threshold concentration required for rainbow trout to ascend the cascade was no more than that. Analyses of thiamine in the muscle of 26 untagged rainbow trout captured in Cayuga Inlet in 2002 showed that 16 fish (62%) had at least 1.0 nmol/g, which was apparently sufficient to support vigorous migration.

Sucrose Penetration and Functionality of Frozen Trout Muscle Affected by Tumbling Conditions

ABSTRACT The effect of tumbling (vacuum pressure: 0, 50, 70 kPa; speed: 7, 14 rpm; time: 30, 60 min) on sucrose penetration into trout muscle was studied when trout chunks were tumbled with 4% (w/w) powdered sucrose. In addition, the cryoprotective action of sucrose in those tumbled muscle was evaluated at -20°C for 24 wk. Sucrose content, expressed as mg/cm2 of exposed area, increased linearly from the center to the periphery of muscle chunks. Vacuum during tumbling and extended tumbling time increased sucrose content of the center portion. Through 24 wk of frozen storage, all tumbling treatments had lower thaw loss and higher gel hardness and cohesiveness than the non-tumbled treatment. Tumbling with 70 kPa vacuum at 7 rpm for 30 min was optimum for sucrose penetration and muscle functionality.

Expression of Four Glutamine Synthetase Genes in the Early Stages of Development of Rainbow Trout (Oncorhynchus mykiss) in Relationship to Nitrogen Excretion

The incorporation of ammonia into glutamine, catalyzed by glutamine synthetase, is thought to be important in the detoxification of ammonia in animals. During early fish development, ammonia is continuously formed as yolk proteins and amino acids are catabolized. We followed the changes in ammonia and urea-nitrogen content, ammonia and urea-nitrogen excretion, glutamine synthetase activity, and mRNA expression of four genes coding for glutamine synthetase (Onmy-GS01-GS04) over 3-80 days post fertilization and in adult liver and skeletal muscle of the rainbow trout (Oncorhynchus mykiss). Both ammonia and urea-nitrogen accumulate before hatching, although the rate of ammonia excretion is considerably higher relative to urea-nitrogen excretion. All four genes were expressed during early development, but only Onmy-GS01 and -GS02 were expressed at appreciable levels in adult liver, and expression was very low in muscle tissue. The high level of expression of Onmy-GS01 and -GS03 prior to hatching corresponded to a linear increase in glutamine synthetase activity. We propose that the induction of glutamine synthetase genes early in development and the subsequent formation of the active protein are preparatory for the increased capacity of the embryo to convert the toxic nitrogen end product, ammonia, into glutamine, which may then be utilized in the ornithine-urea cycle or other pathways.