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Vitamin D3 Enriches Ceramide Content in Exosomes Released by Embryonic Hippocampal Cells
Authors:Carmela Conte  Samuela Cataldi  Cataldo Arcuri  Alessandra Mirarchi  Andrea Lazzarini  Mercedes Garcia-Gil  Tommaso Beccari  Francesco Curcio  Elisabetta Albi
Affiliation:1.Department of Pharmaceutical Sciences, University of Perugia, 06126 Perugia, Italy; (C.C.); (S.C.); (T.B.);2.Department of Medicine and Surgery, University of Perugia, 06126 Perugia, Italy; (C.A.); (A.M.);3.Crabion S.R.L., 06100 Perugia, Italy;4.Department of Biology, Interdepartmental Research Center “Nutraceuticals and Food for Health”, University of Pisa, 56127 Pisa, Italy;5.Department of Medicine, Institute of Clinical Pathology, University of Udine, 33100 Udine, Italy;
Abstract:The release of exosomes can lead to cell–cell communication. Nutrients such as vitamin D3 and sphingolipids have important roles in many cellular functions, including proliferation, differentiation, senescence, and cancer. However, the specific composition of sphingolipids in exosomes and their changes induced by vitamin D3 treatment have not been elucidated. Here, we initially observed neutral sphingomyelinase and vitamin D receptors in exosomes released from HN9.10 embryonic hippocampal cells. Using ultrafast liquid chromatography tandem mass spectrometry, we showed that exosomes are rich in sphingomyelin species compared to whole cells. To interrogate the possible functions of vitamin D3, we established the optimal conditions of cell treatment and we analyzed exosome composition. Vitamin D3 was identified as responsible for the vitamin D receptor loss, for the increase in neutral sphingomyelinase content and sphingomyelin changes. As a consequence, the generation of ceramide upon vitamin D3 treatment was evident. Incubation of the cells with neutral sphingomyelinase, or the same concentration of ceramide produced in exosomes was necessary and sufficient to stimulate embryonic hippocampal cell differentiation, as vitamin D3. This is the first time that exosome ceramide is interrogated for mediate the effect of vitamin D3 in inducing cell differentiation.
Keywords:ceramide   exosome   sphingomyelin   embryonic cell differentiation   vitamin D
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