Pattern of substrate utilization in vascular smooth muscle using 13C isotopomer analysis of glutamate.
Although vascular smooth muscle (VSM) derives the majority of its energy from oxidative phosphorylation, controversy exists concerning which substrates are utilized by the tricarboxylic acid (TCA) cycle. We used 13C isotopomer analysis of glutamate to directly measure the entry of exogenous [13C]glucose and acetate and unlabeled endogenous sources into the TCA cycle via acetyl-CoA. Hog carotid artery segments denuded of endothelium were superfused with 5 mM [1-13C]glucose and 0-5 mM [1,2-13C]acetate at 37 degreesC for 3-12 h. We found that both resting and contracting VSM preferentially utilize [1,2-13C]acetate compared with [1-13C]glucose and unlabeled substrates. The entry of glucose into the TCA cycle (30-60% of total entry via acetyl-CoA) exhibited little change despite alterations in contractile state or acetate concentrations ranging from 0 to 5 mM. We conclude that glucose and nonglucose substrates are important oxidative substrates for resting and contracting VSM. These are the first direct measurements of relative substrate entry into the TCA cycle of VSM during activation and may provide a useful method to measure alterations in VSM metabolism under physiological and pathophysiological conditions.
Allen
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Department of Physiology, University of Missouri, Columbia, Missouri 65212, USA.
Hardin
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D000085:Acetates / Q000378:metabolism; D000105:Acetyl Coenzyme A / Q000378:metabolism; D000818:Animals; D002247:Carbon Isotopes; D002952:Citric Acid Cycle / Q000502:physiology; D005947:Glucose / Q000378:metabolism; D018698:Glutamic Acid / Q000378:metabolism*; D066298:In Vitro Techniques; D009131:Muscle, Smooth, Vascular / Q000378:metabolism* / Q000502:physiology; D009994:Osmolar Concentration; D013552:Swine; D014661:Vasoconstriction / Q000502:physiology
D000085:Acetates; D002247:Carbon Isotopes; D018698:Glutamic Acid; D000105:Acetyl Coenzyme A; D005947:Glucose