Preparation of 13C-labeled ceramide by acetic acid bacteria and its incorporation in mice
| Autor: | Mikiya Kishi, Machiko Iwamoto, Yasukazu Tanaka, Takayuki Kaga, Hiroyuki Fukami, Hatsue Waki, Hideki Tachimoto |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 2010 |
| Předmět: |
Male
Ceramide brain Administration Oral skin epidermis QD415-436 Ceramides Biochemistry Gas Chromatography-Mass Spectrometry chemistry.chemical_compound Acetic acid Mice dihydroceramide Endocrinology Methods stable isotope Animals Acetobacter Acetic acid bacteria Acetic Acid Carbon Isotopes Chromatography biology Epidermis (botany) Sphingosine Cell Biology biology.organism_classification Sphingolipid Mice Inbred C57BL chemistry Isotope Labeling sphingolipid Gas chromatography–mass spectrometry |
| Zdroj: | Journal of Lipid Research, Vol 51, Iss 11, Pp 3389-3395 (2010) |
| ISSN: | 0022-2275 |
| Popis: | 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 dihydroceramide can be incorporated into various tissues, including brain, and metabolized to other sphingolipids. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|