Citrate release by perfused rat hearts: a window on mitochondrial cataplerosis.
Cytosolic citrate is proposed to play a crucial role in substrate fuel selection in the heart. However, little is known about factors regulating the transfer of citrate from the mitochondria, where it is synthesized, to the cytosol. Further to our observation that rat hearts perfused under normoxia release citrate whose (13)C labeling pattern reflects that of mitochondrial citrate (B. Comte, G. Vincent, B. Bouchard, and C. Des Rosiers. J. Biol. Chem. 272: 26117-26124, 1997), we report here data indicating that this citrate release is a specific process reflecting the mitochondrial efflux of citrate, a process referred to as cataplerosis. Indeed, measured rates of citrate release, which vary between 2 and 21 nmol/min, are modulated by the nature and concentration of exogenous substrates feeding acetyl-CoA (fatty acid) and oxaloacetate (lactate plus pyruvate) for the mitochondrial citrate synthase reaction. Such release rates that represent at most 2% of the citric acid cycle flux are in agreement with the activity of the mitochondrial tricarboxylate transporter whose participation is also substantiated by 1) parallel variations in citrate release rates and tissue levels of citrate plus malate, the antiporter, and 2) a lowering of the citrate release rate by 1,2, 3-benzenetricarboxylic acid, a specific inhibitor of the transporter. Taken together, the results from the present study indicate that citrate cataplerosis is modulated by substrate supply, in agreement with the role of cytosolic citrate in fuel partitioning, and occurs, at least in part, through the mitochondrial tricarboxylate transporter.
Vincent
G
G
Department of Biochemistry, University of Montreal, Montreal, Quebec, Canada H3C 3J7.
Comte
B
B
Poirier
M
M
Rosiers
C D
CD
D000105:Acetyl Coenzyme A / Q000378:metabolism; D000818:Animals; D002210:Caprylates / Q000494:pharmacology; D019343:Citric Acid / Q000378:metabolism*; D004734:Energy Metabolism; D008401:Gas Chromatography-Mass Spectrometry; D005947:Glucose / Q000008:administration & dosage; D066298:In Vitro Techniques; D007201:Indicator Dilution Techniques; D007328:Insulin / Q000008:administration & dosage; D007770:L-Lactate Dehydrogenase / Q000378:metabolism; D019344:Lactic Acid / Q000008:administration & dosage; D008297:Male; D008929:Mitochondria, Heart / Q000378:metabolism*; D009206:Myocardium / Q000378:metabolism*; D062907:Oxaloacetic Acid / Q000378:metabolism; D019289:Pyruvic Acid / Q000008:administration & dosage; D051381:Rats; D017207:Rats, Sprague-Dawley
D002210:Caprylates; D007328:Insulin; D019343:Citric Acid; D062907:Oxaloacetic Acid; D019344:Lactic Acid; D000105:Acetyl Coenzyme A; D019289:Pyruvic Acid; D007770:L-Lactate Dehydrogenase; D005947:Glucose; C031492:octanoic acid