Modulation of alpha 6/beta 1 integrin expression during differentiation of F9 murine embryonal carcinoma cells to parietal endoderm.
In vitro differentiation of the murine embryonal carcinoma (EC) cell line F9 parallels that of the early blastocyst, where visceral (VE) and parietal endoderm (PE) diverge from a common precursor, the primitive endoderm. This differentiation pathway is induced by retinoic acid (RA) and dibutyryl cyclicAMP (dcAMP) and is accompanied by progressive and dramatic changes in cell morphology and functions. Within 7 days of treatment the cells organize their cytoskeleton and synthesize large amounts of extracellular matrix proteins, becoming fully differentiated migratory cells; all these changes are likely to involve integrins expression and organization. We have investigated the changes in beta 1 integrin expression, its maturation, and organization on the cell surface in association with alpha 6, during the transition from undifferentiated F9 stem cells to migrating PE cells. By Western blotting and immunoprecipitation we showed a gradual decrease in the amount of the beta 1 subunit on the cell surface and a parallel progressive accumulation of immature protein, indicating that the control of beta 1 expression during F9 cells differentiation occurs first at post-translational level and then at the level of transcription. Moreover, the induction of differentiation produces a marked decrease of alpha 6B and its association to a high molecular weight protein, while alpha 6A level increases. By immunofluorescence we found that upon differentiation there is a relocation of the beta 1 and alpha 6B integrin subunits from cell-cell contacts to focal contacts where they colocalize with vinculin. On the contrary alpha 6A, weakly present in F9 stem cells, is present in the focal contacts of PE cells and along the stress fibers. We suggest different roles for the two alpha 6 isoforms.
Morini
M
M
Unità Transgenici, Università di Genova, Italy.
Piccini
D
D
Barbieri
O
O
Astigiano
S
S
D000818:Animals; D003994:Bucladesine / Q000494:pharmacology; D002451:Cell Compartmentation; D002454:Cell Differentiation / Q000187:drug effects / Q000235:genetics; D002465:Cell Movement; D003599:Cytoskeleton / Q000737:chemistry; D054278:Embryonal Carcinoma Stem Cells; D004707:Endoderm / Q000166:cytology*; D039121:Integrin alpha6beta1; D019012:Integrin beta1 / Q000032:analysis; D016023:Integrins / Q000096:biosynthesis* / Q000235:genetics; D007365:Intercellular Junctions / Q000737:chemistry / Q000502:physiology; D051379:Mice; D014411:Neoplastic Stem Cells / Q000166:cytology*; D015388:Organelles / Q000737:chemistry; D014212:Tretinoin / Q000494:pharmacology; D014407:Tumor Cells, Cultured; D016596:Vinculin / Q000032:analysis
D039121:Integrin alpha6beta1; D019012:Integrin beta1; D016023:Integrins; D016596:Vinculin; D014212:Tretinoin; D003994:Bucladesine