HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS		QUICK SEARCH:   [advanced] Author: Keyword(s): Year:  Vol:  Page:

This Article Full Text Full Text (PDF) All Versions of this Article: 290/5/E1048    most recent 00340.2005v1 Alert me when this article is cited Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Citing Articles Citing Articles via HighWire Citing Articles via Web of Science (8) Citing Articles via Google Scholar Google Scholar Articles by Bederman, I. R. Articles by Previs, S. F. Search for Related Content PubMed PubMed Citation Articles by Bederman, I. R. Articles by Previs, S. F.

INNOVATIVE METHODOLOGY

Novel application of the "doubly labeled" water method: measuring CO₂ production and the tissue-specific dynamics of lipid and protein in vivo

Departments of ¹Nutrition, ²Mathematics, and ³Medicine, Case Western Reserve University, Cleveland, Ohio

Submitted 27 July 2005 ; accepted in final form 14 December 2005

The partitioning of whole body carbon flux between fat and lean compartments affects body composition. We hypothesized that it is possible to simultaneously determine whole body carbon (energy) balance and the dynamics of lipids and proteins in specific tissues in vivo. Growing C57BL/6J mice fed a high-fat low-carbohydrate diet were injected with a bolus of "doubly labeled" water (i.e., ²H₂O and H₂¹⁸O). The rate of CO₂ production was determined from the difference between the elimination rates of ²H and ¹⁸O from body water. The rates of synthesis and degradation of triglycerides extracted from epididymal fat pads and of proteins extracted from heart muscle were determined by mathematically modeling the ²H labeling of triglyceride-bound glycerol and protein-bound alanine, respectively. We found that mice were in positive carbon balance (20% retention per day) and accumulated lipid in epididymal fat pads (9 µmol triglyceride accumulated per day). This is consistent with the fact that mice were studied during a period of growth. Modeling the ²H labeling of triglycerides revealed a substantial rate of lipid breakdown during this anabolic state (equivalent to 25% of the newly synthesized triglyceride). We found equal rates of protein synthesis and breakdown in heart muscle (10% of the pool per day), consistent with the fact that the heart muscle mass did not change. In total, these findings demonstrate a novel application of the doubly labeled water method. Utilization of this approach, especially in unique rodent models, should facilitate studies aimed at quantifying the efficacy of interventions that modulate whole body carbon balance and lipid flux while in parallel determining their impact on (cardiac) muscle protein turnover. Last, the simplicity of administering doubly labeled water and collecting samples allows this method to be used in virtually any laboratory setting.

metabolic regulation; carbon-energy balance; triglyceride turnover; protein turnover; stable isotope tracer kinetics

This article has been cited by other articles:

				H. G. Gasier, S. E. Riechman, M. P. Wiggs, S. F. Previs, and J. D. Fluckey A comparison of 2H2O and phenylalanine flooding dose to investigate muscle protein synthesis with acute exercise in rats Am J Physiol Endocrinol Metab, July 1, 2009; 297(1): E252 - E259. [Abstract] [Full Text] [PDF]

				C. K. Nye, R. W. Hanson, and S. C. Kalhan Glyceroneogenesis Is the Dominant Pathway for Triglyceride Glycerol Synthesis in Vivo in the Rat J. Biol. Chem., October 10, 2008; 283(41): 27565 - 27574. [Abstract] [Full Text] [PDF]

				C. L. Yuan, N. Sharma, D. A. Gilge, W. C. Stanley, Y. Li, M. Hatzoglou, and S. F. Previs Preserved protein synthesis in the heart in response to acute fasting and chronic food restriction despite reductions in liver and skeletal muscle Am J Physiol Endocrinol Metab, July 1, 2008; 295(1): E216 - E222. [Abstract] [Full Text] [PDF]

				E. Belloto, F. Diraison, A. Basset, G. Allain, P. Abdallah, and M. Beylot Determination of protein replacement rates by deuterated water: validation of underlying assumptions Am J Physiol Endocrinol Metab, May 1, 2007; 292(5): E1340 - E1347. [Abstract] [Full Text] [PDF]