A Comparative Study of Stable Isotope Tracing of Hepatic Metabolism Using [U-13C]Propionate and [U-13C]Glutamine

Clayton Olinger

Authors:  Clayton Olinger, Mukundan Ragavan, Anthony Giacalone, and Matthew Merritt

Faculty Mentor:  Dr. Matthew E. Merritt

College:  College of Medicine

Abstract

The use of isotopic tracers is an effective way to assess metabolic turnover, as well as various pathophysiologic perturbations of normal metabolic flux. The isolation and the perfusion of mice livers allow for measurements of hepatic citric acid cycle flux, as well as gluconeogenesis by tracking these isotopes in 2H and 13C NMR spectroscopy. Perfusions are performed by making an incision in the abdomen of the mice, allowing for a thin tube to be put into the portal vein of the liver. This liver is removed, cleaned and placed in a glass column where it is be perfused with a Krebs buffer, mixed fatty acids, lactate and pyruvate. These perfusions allow the liver to be metabolically active outside of the mouse. [U-13C]propionate, a labeled odd-chain fatty acid, has been used in the past to measure the oxidative capacity of the liver. Using NMR spectroscopy, we probe hepatic metabolism using [U-13C]glutamine and compare the results to the [U-13C]propionate method. Two different concentrations of [U-13C]glutamine are evaluated for its ability to report on gluconeogenesis and citric acid cycle turnover. Even at the lower concentration of 100 µM, glutamine can accurately report citric acid flux and gluconeogenesis activity in perfused mice livers.

Poster Pitch

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Poster

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Clayton Olinger
Clayton Olinger (@guest_1760)
1 year ago

Hello All,

Thank you for coming to my page to check out some of the research we are doing in the department of biochemistry. If you have any questions, need me to clarify anything, or just have any other other comments about the material in the video or in the poster, don’t be afraid to comment.

Go gators!

Clayton Olinger
Clayton Olinger (@guest_2996)
1 year ago

Here is the zoom link I will be on from 3-4 if you’d rather ask a question through there.

https://us04web.zoom.us/j/864389891

Ivanna di Lorenzo
Ivanna di Lorenzo (@guest_3552)
1 year ago

Good job on your presentation and poster! It looks amazing!

Clayton Olinger
Clayton Olinger (@guest_6068)
Reply to  Ivanna di Lorenzo
1 year ago

Thanks Ivanna!