Determination of muscle protein synthesis rates in fish using 2H2O and 2H NMR analysis of alanine

Marques, Cátia; Viegas, Filipa; Rito, João; Jones, John G.; Viegas, Iván

doi:10.1016/j.ab.2016.07.011

Cited by 5 publications

(5 citation statements)

References 21 publications

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“…The same protocol recommends formate as an internal standard as opposed to TSP, however, endogenous formate was observed in preliminary plasma samples. Pyrazine was therefore chosen as an internal standard at a concentration of 0.33 mM [ 31 , 32 ]. The pH was adjusted to 7.4.…”

Section: Methodsmentioning

confidence: 99%

Determining bacterial and host contributions to the human salivary metabolome

Gardner

Parkes

et al. 2019

Journal of Oral Microbiology

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Background: Salivary metabolomics is rapidly advancing. Aim and methods: To determine the extent to which salivary metabolites reflects host or microbial metabolic activity whole-mouth saliva (WMS), parotid saliva (PS) and plasma collected contemporaneously from healthy volunteers were analysed by 1 H-NMR spectroscopy. Spectra underwent principal component analysis and k-means cluster analysis and metabolite quantification. WMS samples were cultured on both sucrose and peptide-enriched media. Correlation between metabolite concentration and bacterial load was assessed. Results: WMS contained abundant short-chain fatty acids (SCFAs), which were minimal in PS and plasma. WMS spectral exhibited greater inter-individual variation than those of PS or plasma (6.7 and 3.6 fold, respectively), likely reflecting diversity of microbial metabolomes. WMS bacterial load correlated strongly with SCFA levels. Additional WMS metabolites including amines, amino acids and organic acids were positively correlated with bacterial load. Lactate, urea and citrate appeared to enter WMS via PS and the circulation. Urea correlated inversely with WMS bacterial load. Conclusions: Oral microbiota contribute significantly to the WMS metabolome. Several WMS metabolites (lactate, urea and citrate) are derived from the host circulation. WMS may be particularly useful to aid diagnosis of conditions reflective of dysbiosis. WMS could also complement other gastrointestinal fluids in future metabolomic studies. ARTICLE HISTORY

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Section: Methodsmentioning

confidence: 99%

Determining bacterial and host contributions to the human salivary metabolome

Gardner

Parkes

et al. 2019

Journal of Oral Microbiology

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“…In vitro studies, namely the infusion of adipocytes with 14 C-glucose (Bou et al, 2016) have also contributed to a better understanding this subject. Instead of following the metabolic fate of a single labeled substrate, the stable isotope deuterium ( 2 H), presented as deuterated water ( 2 H2O) in fish tanks, rapidly equilibrates with fish body water (Viegas et al, 2011) and gets widely incorporated in newly synthesized metabolites such as alanine (Marques et al, 2016), glucose or glycogen (Viegas et al, 2015). Similarly, 2 H gets incorporated into different sites of the triacylglycerol (TAG) molecule following well-defined metabolic steps from which estimations for de novo lipogenesis (DNL) and glycerol turnover can be derived (Viegas et al, 2016).…”

Section: Introductionmentioning

confidence: 99%

Impact of dietary starch on extrahepatic tissue lipid metabolism in farmed European (Dicentrarchus labrax) and Asian seabass (Lates calcarifer)

Viegas

Trenkner

Rito

et al. 2019

Comparative Biochemistry and Physiology Part A: Molecular &

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In aquaculture, there is high interest in substituting marine-derived with vegetablebased ingredients as energy source. Farmed carnivorous fish under high carbohydrate diets tend to increase adiposity but it remains unclear if this happens by increased lipid retention/accumulation, promotion of lipogenic pathways, or both. In order to determine the response of extrahepatic tissue to dietary starch, European (Dicentrarchus labrax) and Asian (Lates calcarifer) seabass were fed a control (low starch; LS) or experimental (high starch; HS) diet, for at least 21 days and then transferred for 6 days to saltwater enriched with deuterated water 2 H2O. Incorporation of 2 H-labelling follows well-defined metabolic steps, and analysis of triacylglycerols (TAG) 2 H-enrichment by 2 H-NMR allowed evaluation of de novo lipogenesis (DNL) in muscle and visceral adipose tissue (VAT). Fractional synthetic rates for TAGbound fatty acids and glycerol were quantified separately providing a detailed lipogenic profile. The FA profile differed substantially between muscle and VAT in both species, but their lipogenic fluxes revealed even greater differences. In European seabass, HS promoted DNL of TAG-bound FA, in muscle and VAT. High 2 Henrichment also found in muscle TAG-bound glycerol was indicative of its role on lipid cycling. In Asian seabass, HS had no effect on muscle FA composition and lipogenic flux, with no 2 H-enriched TAG being detected. VAT on the other hand revealed a strong enhancement of DNL in HS-fed fish along with high TAG-bound glycerol cycling. This study consolidated the use of 2 H2O as tracer for fish lipid metabolism in different tissues, under different dietary conditions and suitable to use in different fish models.

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“…Until recently, however, a deuterated ‘heavy’ water ( 2 H 2 O) technique that linearly equilibrates biosynthetic pathways, to enable steady state assessment of metabolism between sink tissues and plasma was developed in other teleosts 267,268 . 2 H 2 O eliminates issues with tank water and unlabelled precursor dilution, and enables the segregation of glucose into gluconeogenic and glycogenic sources 271 .…”

Section: Research Themes In Salmon Metabolomicsmentioning

confidence: 99%

“…263 Metabolic flux can thus be used to aid the development of novel aquafeed with the target to sustain growth, improve fillet quality, promote animal health and wellbeing 249,264 as previously demonstrated with dietary starch, 248,249,265,266 glycerol 243 and proteins in other teleosts. [266][267][268] For several reasons however, metabolic flux analysis remain unexploited in salmonid aquaculture nutrition, potentially due to the recent adoption of metabolomics in aquaculture research. 264,269 Furthermore, the unpredictable nature of organ systems supplied by a single blood stream limits modelling of a specific tissue site in a living fish due to cross-metabolism of a labelled tracer.…”

Section: Areas For Improvement In Salmonid Metabolomicsmentioning

confidence: 99%

“…243 Using teleost D. labrax, the application of 2 H 2 O has been extensively demonstrated to practically study sink-specific biosynthetic pathways for lipids, 248 carbohydrates 271,272 and proteins 267,268 and maybe adapted in salmonid nutritional studies using 2 H-NMR and high-resolution MS metabolomics. 267,271 Interestingly, innovative nanoscale secondary ion mass spectrometry (NanoSIMS) and microfluidic NMR devices have emerged and will likely facilitate the uptake of metabolic flux analysis in salmon aquaculture research at tissue and cellular levels. 34,74,273 Biochemical characterisation of farmed salmon will help to mitigate the long-held belief that wild fish is of superior quality to farmed counterparts.…”

Section: Areas For Improvement In Salmonid Metabolomicsmentioning

confidence: 99%

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Metabolomics in salmonid aquaculture research: Applications and future perspectives

Lulijwa

Young

2021

Reviews in Aquaculture

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Salmonids are the third major farmed finfish species after carps and tilapines, and thus contribute to global food fish production. However, as the aquaculture industry continues to grow, several challenges have emerged. Left unchecked, current problems will hinder aquaculture development. Metabolomics is one of the powerful biotechnological tools that will contribute to solving some of the pressing problems. This review aims to summarise findings and identify gaps from studies that used metabolomics in farmed salmon research. We extracted methodological information for comparison, highlighted analytical platform usages and identified most studied salmonids and sample types. We reviewed key articles to highlight the latest research themes.From the identified research gaps, future perspectives regarding potential use of metabolomics to solve issues in ecotoxicology, thermotolerance, nutrition, post-harvest quality, health and disease, and husbandry practices are provided. The survey also highlighted improvements in execution of metabolomics protocols in aquaculture.

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Determination of muscle protein synthesis rates in fish using 2H2O and 2H NMR analysis of alanine

Cited by 5 publications

References 21 publications

Determining bacterial and host contributions to the human salivary metabolome

Determining bacterial and host contributions to the human salivary metabolome

Impact of dietary starch on extrahepatic tissue lipid metabolism in farmed European (Dicentrarchus labrax) and Asian seabass (Lates calcarifer)

Metabolomics in salmonid aquaculture research: Applications and future perspectives

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