Isotopomics: a top-down systems biology approach for understanding dynamic metabolism in rats using [1,2-(13)C(2)] acetate.
Isotope labeled tracers are commonly used to quantify the turnover rates of various metabolic intermediates and yield information regarding physiological regulation. Studies often only consider either one nutritional state (fasted or fed) and/or one question (e.g., measure of lipid or protein turnover). In this article, we consider a novel application combining the global approach of metabonomics with widespread stable isotope labeling as a way of being able to map metabolism in open mammalian systems, an approach we call "isotopomics". A total of 45 15-week-old male Zucker rats were administrated different amounts (from 0.5 to 8 mmol/kg) of sodium [1,2-(13)C(2)] acetate. Plasma samples taken at 1, 4, and 24 h were analyzed with (13)C nuclear magnetic resonance (NMR) and gas chromatography/mass spectrometry (GC/MS) to measure (13)C isotopic enrichment of 39 plasma metabolites across a wide range of compound classes (amino acids, short-chain fatty acids, lactate, glucose, and free fatty acids). Isotopic enrichment from 0.1-7.1 mole percent excess (MPE) for the highest dose could be reliably measured in 16 metabolites, and the kinetics of their (13)C isotopic enrichment are reported. Clustering metabolites based on (13)C kinetic curves enabled highlighting of time dependent patterns of (13)C distribution through the key metabolic pathways. These kinetic and quantitative data were reported into a biochemical map. This type of isotopomic approach for mapping dynamic metabolism in an open system has great potential for advancing our mechanistic knowledge of how different interventions and diseases can impact the metabolic response of animals and humans.
Godin
Jean-Philippe
JP
Nestlé Research Centre, Vers-Chez-les-Blanc, 1000 Lausanne 26, Switzerland. Jean-Philippe.Godin@rdls.nestle.com
Ross
Alastair B
AB
Rezzi
Serge
S
Poussin
Carine
C
Martin
Francois-Pierre
FP
Fuerholz
Andreas
A
Cléroux
Marilyn
M
Mermoud
Anne-France
AF
Tornier
Lionel
L
Arce Vera
Francia
F
Pouteau
Etienne
E
Ramadan
Ziad
Z
Kochhar
Sunil
S
Fay
Laurent-Bernard
LB
D000818:Animals; D002247:Carbon Isotopes / Q000378:metabolism; D008401:Gas Chromatography-Mass Spectrometry / Q000379:methods*; D007700:Kinetics; D009682:Magnetic Resonance Spectroscopy / Q000379:methods*; D008297:Male; D055432:Metabolomics; D015999:Multivariate Analysis; D051381:Rats; D019346:Sodium Acetate / Q000097:blood / Q000378:metabolism*; D013997:Time Factors
D002247:Carbon Isotopes; D019346:Sodium Acetate