IQUIFIB   02644
INSTITUTO DE QUIMICA Y FISICOQUIMICA BIOLOGICAS "PROF. ALEJANDRO C. PALADINI"
Unidad Ejecutora - UE
congresos y reuniones científicas
Título:
Importance of the Sphingolipid Metabolism in the Acquisition of Final MDCK Differentiation
Autor/es:
PESCIO LUCILA; NICOLÁS OCTAVIO FAVALE; NORMA STERIN-SPEZIALE
Lugar:
Boston
Reunión:
Congreso; Experimental Biology 2015; 2015
Institución organizadora:
American Society of Biochemistry and Molecular Biology (ASBMB)
Resumen:
The final stage of epithelial cell differentiation involves the outgrowth of primary cilium and the development of mature apical membrane. We have demonstrated that glycosphingolipid (GSL) synthesis is essential for MDCK cell differentiation induced by hypertonicity. Sphingolipid metabolism includes ¨de novo ¨and the ¨salvage pathway¨. The aim of this study was to evaluate the role of sphingolipid metabolism in MDCK cell differentiation.Confluent MDCK cells subjected to hypertonic media for 72h in the presence or absence of serine palmitoil transferase (SPT), ceramide synthases (CerS), glucosylceramide synthase (GCerS), sphingomyelin synthase (SMS) and acid sphingomyelinase (aSMase) inhibitors. Cilium was revealed by immunofluorescence for acetylated tubulin and anti-gp135 as marker of apical membrane. Hypertonicity-induced cilium formation was impaired by D-PDMP and D609 also affected apical membrane maturation. aSMase inhibition evoked a highest impairment of cell differentiation. Both SPT and CerS synthase inhibition altered cell phenotype. Results demonstrate that, MDCK cells cannot differentiate in the presence of D-PDMP and D609, indicating that GCer and SM synthesis are essential for primary cilium formation. Results also showed that both SM and GSL involved in MDCK differentiation must come from the salvage pathway, having endolysosomal ceramide as direct substrate for their synthesis. Supported by grants from the CONICET and the University of Buenos Aires.