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Hyperpolarized [1- 13 C]Acetyl-l-Carnitine Probes Tricarboxylic Acid Cycle Activity In Vivo .
ACS Sensors 2023 August 15
Mitochondrial oxidative phosphorylation (OXPHOS) is sensitive to a variety of biological factors, and dysregulated OXPHOS is observed during the development of numerous pathological conditions. ATP production via OXPHOS is intrinsically dependent on the availability of acetyl-coenzyme A (CoA), which can enter the tricarboxylic acid (TCA) cycle to drive the oxidative pathway. Acetyl-l-carnitine (ALCAR) is an interchangeable endogenous source of acetyl-CoA, and therefore, ALCAR-derived probes are uniquely positioned for the assessment of OXPHOS. In this report, we develop hyperpolarized (HP) [1-13 C]ALCAR as a noninvasive probe to investigate cardiac TCA cycle activity in vivo . We initially synthesized the isotopically labeled substrate and demonstrated that the 13 C nucleus maintained a suitable T 1 value (50.1 ± 0.8 s at 3 T) and polarization levels (21.3 ± 5.3%) to execute in vivo metabolic measurements. HP [1-13 C]ALCAR was employed for cardiac analyses of OXPHOS in rats under fed and fasted conditions. [5-13 C]Glutamate was successfully detected, and the metabolite was used to analyze the TCA cycle activity in both nutritional states. These assessments were compared to analogous experiments with the HP [1-13 C]pyruvate. Our report represents the first study to demonstrate that HP methods using [1-13 C]ALCAR enable direct analyses of mitochondrial function and TCA cycle activity, which are fundamental to cardiac cell homeostasis.
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