|
|
||||||||
Correspondence to:
Nicholas D. Oakes
We describe a method for assessing tissue-specific plasma free fatty acid (FFA) utilization in vivo using a non-ß-oxidizable FFA analog, [9,10-3H]-(R)-2-bromopalmitate (3H-R-BrP). Ideally 3H-R-BrP would be transported in plasma, taken up by tissues and activated by the enzyme acyl-CoA synthetase (ACS) like native FFA, but then 3H-labeled metabolites would be trapped. In vitro we found that 2-bromopalmitate and palmitate compete equivalently for the same ligand binding sites on albumin and intestinal fatty acid binding protein, and activation by ACS was stereoselective for the R-isomer. In vivo, oxidative and non-oxidative FFA metabolism was assessed in anesthetized Wistar rats by infusing, over 4 min, a mixture of 3H-R-BrP and [U-14C] palmitate (14C-palmitate). Indices of total FFA utilization (R*f) and incorporation into storage products (Rfs') were defined, based on tissue concentrations of 3H and 14C, respectively, 16 min after the start of tracer infusion. R*f, but not Rfs', was substantially increased in contracting (sciatic nerve stimulated) hindlimb muscles compared with contralateral non-contracting muscles. The contraction-induced increases in R*f were completely prevented by blockade of ß-oxidation with etomoxir. These results verify that 3H-R-BrP traces local total FFA utilization, including oxidative and non-oxidative metabolism. Separate estimates of the rates of loss of 3H activity indicated effective 3H metabolite retention in most tissues over a 16-min period, but appeared less effective in liver and heart.
In conclusion, simultaneous use of 3H-R-BrP and [14C]palmitate tracers provides a new useful tool for in vivo studies of tissue-specific FFA transport, utilization and metabolic fate, especially in skeletal muscle and adipose tissue.Oakes, N. D., A. Kjellstedt, G-B. Forsberg, T. Clementz, G. Camejo, S. M. Furler, E. W. Kraegen, M. Ölwegård-Halvarsson, A. B. Jenkins, and B. Ljung. Development and initial evaluation of a novel method for assessing tissue-specific plasma free fatty acid utilization in vivo using (R)-2-bromopalmitate tracer. J. Lipid Res. 1999. 40: 11551169.
Supplementary key words:
metabolism, turnover, analog, muscle, liver, heart, adipose tissue, kinetics
Copyright © 1999 by Lipid Research, Inc.
Methods
Development and initial evaluation of a novel method for assessing tissue-specific plasma free fatty acid utilization in vivo using (R)-2-bromopalmitate tracer
Nicholas D. Oakesa,
Ann Kjellstedta,
Gun-Britt Forsberga,
Tony Clementza,
Germán Camejoa,
Stuart M. Furlerb,
Edward W. Kraegenb,
Maria Ölwegård-Halvarssona,
Arthur B. Jenkinsb, and
Bengt Ljunga
a Department of Pharmacology, Astra-Hässle AB, S-431 83 Mölndal, Sweden
b Garvan Institute of Medical Research, Sydney, New South Wales, Australia
![]()
CiteULike
Complore
Connotea
Del.icio.us
Digg
Reddit
Technorati What's this?
This article has been cited by other articles:
![]() |
C. Koutsari, D. A. Dumesic, B. W. Patterson, S. B. Votruba, and M. D. Jensen Plasma Free Fatty Acid Storage in Subcutaneous and Visceral Adipose Tissue in Postabsorptive Women Diabetes, May 1, 2008; 57(5): 1186 - 1194. [Abstract] [Full Text] [PDF] |
||||
![]() |
E. Diakogiannaki, S. Dhayal, C. E Childs, P. C Calder, H. J Welters, and N. G Morgan Mechanisms involved in the cytotoxic and cytoprotective actions of saturated versus monounsaturated long-chain fatty acids in pancreatic {beta}-cells J. Endocrinol., August 1, 2007; 194(2): 283 - 291. [Abstract] [Full Text] [PDF] |
||||
![]() |
N. M. Borradaile, X. Han, J. D. Harp, S. E. Gale, D. S. Ory, and J. E. Schaffer Disruption of endoplasmic reticulum structure and integrity in lipotoxic cell death J. Lipid Res., December 1, 2006; 47(12): 2726 - 2737. [Abstract] [Full Text] [PDF] |
||||
![]() |
H. A. Parkes, E. Preston, D. Wilks, M. Ballesteros, L. Carpenter, L. Wood, E. W. Kraegen, S. M. Furler, and G. J. Cooney Overexpression of acyl-CoA synthetase-1 increases lipid deposition in hepatic (HepG2) cells and rodent liver in vivo Am J Physiol Endocrinol Metab, October 1, 2006; 291(4): E737 - E744. [Abstract] [Full Text] [PDF] |
||||
![]() |
K. Fosgerau, C. Fledelius, K. E Pedersen, J. B Kristensen, J. R Daugaard, M. A Iglesias, E. W Kraegen, and S. M Furler Oral administration of glucose promotes intracellular partitioning of fatty acid toward storage in white but not in red muscle. J. Endocrinol., September 1, 2006; 190(3): 651 - 658. [Abstract] [Full Text] [PDF] |
||||
![]() |
N. D. Oakes, P. Thalen, E. Aasum, A. Edgley, T. Larsen, S. M. Furler, B. Ljung, and D. Severson Cardiac metabolism in mice: tracer method developments and in vivo application revealing profound metabolic inflexibility in diabetes Am J Physiol Endocrinol Metab, May 1, 2006; 290(5): E870 - E881. [Abstract] [Full Text] [PDF] |
||||
![]() |
A. D. Hafstad, G. H. Solevag, D. L. Severson, T. S. Larsen, and E. Aasum Perfused hearts from Type 2 diabetic (db/db) mice show metabolic responsiveness to insulin Am J Physiol Heart Circ Physiol, May 1, 2006; 290(5): H1763 - H1769. [Abstract] [Full Text] [PDF] |
||||
![]() |
M. A. Iglesias, S. M. Furler, G. J. Cooney, E. W. Kraegen, and J.-M. Ye AMP-Activated Protein Kinase Activation by AICAR Increases Both Muscle Fatty Acid and Glucose Uptake in White Muscle of Insulin-Resistant Rats In Vivo Diabetes, July 1, 2004; 53(7): 1649 - 1654. [Abstract] [Full Text] [PDF] |
||||
![]() |
B. D. Hegarty, S. M. Furler, N. D. Oakes, E. W. Kraegen, and G. J. Cooney Peroxisome Proliferator-Activated Receptor (PPAR) Activation Induces Tissue-Specific Effects on Fatty Acid Uptake and Metabolism in Vivo--A Study Using the Novel PPAR{alpha}/{gamma} Agonist Tesaglitazar Endocrinology, July 1, 2004; 145(7): 3158 - 3164. [Abstract] [Full Text] [PDF] |
||||
![]() |
B. P. Atshaves, S. M. Storey, and F. Schroeder Sterol carrier protein-2/sterol carrier protein-x expression differentially alters fatty acid metabolism in L cell fibroblasts J. Lipid Res., September 1, 2003; 44(9): 1751 - 1762. [Abstract] [Full Text] [PDF] |
||||
![]() |
B. Teusink, P. J. Voshol, V. E.H. Dahlmans, P. C.N. Rensen, H. Pijl, J. A. Romijn, and L. M. Havekes Contribution of Fatty Acids Released From Lipolysis of Plasma Triglycerides to Total Plasma Fatty Acid Flux and Tissue-Specific Fatty Acid Uptake Diabetes, March 1, 2003; 52(3): 614 - 620. [Abstract] [Full Text] [PDF] |
||||
![]() |
B. D. Hegarty, G. J. Cooney, E. W. Kraegen, and S. M. Furler Increased Efficiency of Fatty Acid Uptake Contributes to Lipid Accumulation in Skeletal Muscle of High Fat-Fed Insulin-Resistant Rats Diabetes, May 1, 2002; 51(5): 1477 - 1484. [Abstract] [Full Text] [PDF] |
||||
![]() |
M. Cnop, J. C. Hannaert, A. Hoorens, D. L. Eizirik, and D. G. Pipeleers Inverse Relationship Between Cytotoxicity of Free Fatty Acids in Pancreatic Islet Cells and Cellular Triglyceride Accumulation Diabetes, August 1, 2001; 50(8): 1771 - 1777. [Abstract] [Full Text] [PDF] |
||||
| HOME | HELP | FEEDBACK | SUBSCRIPTIONS | ARCHIVE | SEARCH | TABLE OF CONTENTS |
| All ASBMB Journals | Journal of Biological Chemistry |
| Molecular and Cellular Proteomics | ASBMB Today |