Preparation of (13)C-labeled ceramide by acetic acid bacteria and its incorporation in mice.

Journal of Lipid Research
Hiroyuki FukamiYasukazu Tanaka

Abstract

We prepared 2-hydroxypalmitoyl-sphinganine (dihydroceramide) labeled with a stable isotope by culturing acetic acid bacteria with (13)C-labeled acetic acid. The GC/MS spectrum of the trimethylsilyl derivative of (13)C-labeled dihydroceramide gave molecular ions with an increased mass of 12-17 Da over that of nonlabeled dihydroceramide. The fragment ions derived from both sphinganine base and 2-hydroxypalmitate were confirmed to be labeled with the stable isotope in the spectrum. Therefore, (13)C-labeled dihydroceramide can be an extremely useful tool for analyzing sphingolipid metabolism. The purified [(13)C]dihydroceramide was administered orally to mice for 12 days, and the total sphingoid base fractions in various tissues were analyzed by GC/MS. The spectrum patterns specific to (13)C-labeled sphingoids were detected in the tissues tested. Sphinganine pools in skin epidermis, liver, skeletal muscle, and synapse membrane in brain were replaced by [(13)C]sphinganine at about 4.5, 4.0, 1.0, and 0.3%, respectively. Moreover, about 1.0% of the sphingosine pool in the liver was replaced by [(13)C]sphingosine, implying that exogenous dihydroceramide can be converted to sphingosine. These results clearly indicate that ingested dihyd...Continue Reading

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Citations

Apr 9, 2013·Bioscience, Biotechnology, and Biochemistry·Chinatsu KawadaYoshihiro Nomura
Oct 7, 2014·Biochimica Et Biophysica Acta·S Rodriguez-CuencaA Vidal-Puig
Mar 7, 2018·American Journal of Physiology. Endocrinology and Metabolism·Ying ChenDavid G McLaren
Oct 21, 2020·Canadian Journal of Microbiology·Ethendhar RajendiranVanu Ramprasath
Jul 11, 2020·Journal of Lipid Research·Min-Ting LeeElizabeth L Johnson
Feb 9, 2021·Journal of Lipid Research·Min-Ting LeeElizabeth L Johnson

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