IQUIFIB   02644
INSTITUTO DE QUIMICA Y FISICOQUIMICA BIOLOGICAS "PROF. ALEJANDRO C. PALADINI"
Unidad Ejecutora - UE
congresos y reuniones científicas
Título:
COORDINATION BETWEEN SPHINGOLIPIDS AND PROTEINS REQUIRED FOR PRIMARY CILIOGENESIS
Autor/es:
MARíA S TULINO; NICOLáS O FAVALE; BRUNO J SANTACREU; JOSE LUIS E MONTI; LUCILA G PESCIO; CARLOS H PAVáN; NORMA B STERIN-SPEZIALE
Lugar:
Buenos Aires
Reunión:
Congreso; Reunión conjunta de sociedades de biociencias; 2017
Institución organizadora:
SAIB-SAIC-SAI-SAA-SAB-SAB-SAFE-SAFIS-SAH-SAP
Resumen:
The final stage in the process of epithelial celldifferentiation isthe outgrowth of a primary cilium from the apicalsurface. We havepreviously described that extracellular hypertonicityinduces thedifferentiation of MDCK cells by the regulation of thesphingolipidmetabolism. The aim of this work was to investigatethe ciliogenesisinduced by hypertonicity in MDCK cells and the role ofglycosphingolipidsin the process. Confluent MDCK cells were culturedunderisotonicity (150 mM NaCl, control) or hypertonicity(300 mM NaCl)for 48 h in the presence or absence of D-PDMP, aglucosylceramidesynthase inhibitor. The presence of primary cilium wasdetected inMDCK cells stable expressing á-tubulin?GFP subjected to hypertonicity.For the immunofluorescence assays, cells were stainedwithacetylated tubulin to visualize primary cilium, andco-stained withIFT20 or Rab8, proteins required for vesicle-mediatedtraffickingpathway from the Golgi to the base of the cilium.Isotonicity-culturedcells did not show cilium formation and IFT20 and Rab8appeareddiffusely distributed. Cells cultured underhypertonicity displayed primarycilium and the number of ciliated cells significantlydecreasedwhen the cells were subjected to hypertonicity in thepresence ofD-PDMP. Cells cultured under hypertonicity showedIFT20 with aGolgi ? like distribution and perinuclear Rab8, butwhen cells weretreated with D-PDMP the distribution of these proteinswas diffuse.A proteomic analysis performed by mass spectrometry ofIFT20+vesicles immune-isolated from cells cultured underhypertonicity revealedthe presence of galectin 3, a protein with animportant rolein apical sorting that associates with the primarycilium. In conjunction,these results suggest that hypertonicity inducesciliogenesis bydisplaying a protein machinery that is correctlyassembled in thepresence of glycosphingolipids.