Changes in ceramide metabolism are essential in Madin-Darby canine kidney cell differentiation

Ceramides (Cers) and complex sphingolipids with defined acyl chain lengths play important roles in numerous cell processes. Six Cer synthase (CerS) isoenzymes (CerS1–6) are the key enzymes responsible for the production of the diversity of molecular species. In this study, we investigated the change...

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Published in:Journal of lipid research 2017-07, Vol.58 (7), p.1428-1438
Main Authors: Pescio, Lucila Gisele, Santacreu, Bruno Jaime, Lopez, Vanina Gisela, Paván, Carlos Humberto, Romero, Daniela Judith, Favale, Nicolás Octavio, Sterin-Speziale, Norma Beatriz
Format: Article
Language:English
Subjects:
Amitriptyline
Animals
Cell Differentiation
Ceramide
ceramide synthase
Ceramides - metabolism
Dogs
Gene Expression Regulation, Enzymologic
glycolipids
Glycosphingolipids
Hypertonicity
Isoenzymes
Lipid metabolism
Madin Darby Canine Kidney Cells
mass spectrometry
Metabolism
Oxidoreductases - genetics
Species
Sphingolipids
Sphingomyelin phosphodiesterase
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Summary:Ceramides (Cers) and complex sphingolipids with defined acyl chain lengths play important roles in numerous cell processes. Six Cer synthase (CerS) isoenzymes (CerS1–6) are the key enzymes responsible for the production of the diversity of molecular species. In this study, we investigated the changes in sphingolipid metabolism during the differentiation of Madin-Darby canine kidney (MDCK) cells. By MALDI TOF TOF MS, we analyzed the molecular species of Cer, glucosylceramide (GlcCer), lactosylceramide (LacCer), and SM in nondifferentiated and differentiated cells (cultured under hypertonicity). The molecular species detected were the same, but cells subjected to hypertonicity presented higher levels of C24:1 Cer, C24:1 GlcCer, C24:1 SM, and C16:0 LacCer. Consistently with the molecular species, MDCK cells expressed CerS2, CerS4, and CerS6, but with no differences during cell differentiation. We next evaluated the different synthesis pathways with sphingolipid inhibitors and found that cells subjected to hypertonicity in the presence of amitriptyline, an inhibitor of acid sphingomyelinase, showed decreased radiolabeled incorporation in LacCer and cells did not develop a mature apical membrane. These results suggest that hypertonicity induces the endolysosomal degradation of SM, generating the Cer used as substrate for the synthesis of specific molecular species of glycosphingolipids that are essential for MDCK cell differentiation.
ISSN:0022-2275
1539-7262
DOI:10.1194/jlr.M076349