RESUMO
Epithelial cell shape change is a pivotal driving force for morphogenesis of complex three-dimensional architecture. However, molecular mechanisms triggering shape changes of epithelial cells in the course of growth and differentiation have not been entirely elucidated. Grhl3 plays a crucial role as a downstream transcription factor of Wnt/ß-catenin in epidermal differentiation. Here, we show Grhl3 induced large, mature epidermal cells, enriched with actomyosin networks, from embryoid bodies in vitro. Such epidermal cells were apparently formed by the simultaneous activation of canonical and non-canonical Wnt signaling pathways. A nuclear transcription factor, GRHL3 is localized in the cytoplasm and cell membrane during epidermal differentiation. Subsequently, such extranuclear GRHL3 is essential for the membrane-associated expression of VANGL2 and CELSR1. Cytoplasmic GRHL3, thereby, allows epidermal cells to acquire mechanical properties for changes in epithelial cell shape. Thus, we propose that cytoplasmic localization of GRHL3 upon epidermal differentiation directly triggers epithelial morphogenesis.
Assuntos
Diferenciação Celular , Forma Celular , Citoplasma/metabolismo , Proteínas de Ligação a DNA/metabolismo , Epiderme/metabolismo , Células Epiteliais/citologia , Células Epiteliais/metabolismo , Morfogênese , Fatores de Transcrição/metabolismo , Alelos , Animais , Biomarcadores/metabolismo , Polaridade Celular , Citoesqueleto/metabolismo , Corpos Embrioides/metabolismo , Células Epidérmicas/metabolismo , Camundongos , Camundongos Transgênicos , Modelos Biológicos , Mutação/genética , Células NIH 3T3 , Neurulação , Via de Sinalização WntRESUMO
To understand genetic programs controlling mammalian central nervous system (CNS) development, we have identified one transgene-inserted mutation, which showed embryonic lethality during neurulation. Determination of the transgene integration site and rescue experiments revealed that the Brd2 gene, whose products specifically bind acetylated histone H4 and can mediate transcription, was the cause of this mutation. Expression studies with specific markers demonstrated that cell cycle progression was accelerated and neuronal differentiation as well as cell cycle exit were impaired in Brd2-deficient neruoepithelial cells. To investigate whether Brd2 regulates neuronal differentiation through a E2F1 transcriptional factor, which directly binds Brd2 and controls genes expression for cell cycle progression and exit, we analyzed Brd2;E2F1 double mutant phenotypes and, consequently found that abnormalities in neuronal differentiation and cell cycle progression due to Brd2-deficiency were restored by removing the E2F1 gene. These findings suggest that Brd2 is required for cell cycle exit and neuronal differentiation of neuroepithelial cells through the E2F1 pathway during mouse CNS development.
Assuntos
Ciclo Celular/fisiologia , Sistema Nervoso Central/crescimento & desenvolvimento , Fator de Transcrição E2F1/metabolismo , Células Neuroepiteliais/citologia , Neurogênese/fisiologia , Neurônios/citologia , Proteínas Serina-Treonina Quinases/fisiologia , Alelos , Animais , Ciclo Celular/genética , Diferenciação Celular , Sistema Nervoso Central/metabolismo , Proteínas Cromossômicas não Histona , Fator de Transcrição E2F1/genética , Camundongos , Camundongos Transgênicos , Neurogênese/genética , Proteínas Serina-Treonina Quinases/genética , Fatores de TranscriçãoRESUMO
Heparan sulfate (HS) proteoglycans modulate the activity of multiple growth factors on the cell surface and extracellular matrix. However, it remains unclear how the HS chains control the movement and reception of growth factors into targeted receiving cells during mammalian morphogenetic processes. Here, we found that HS-deficient Ext2 null mutant mouse embryos fail to respond to fibroblast growth factor (FGF) signaling. Marker expression analyses revealed that cell surface-tethered HS chains are crucial for local retention of FGF4 and FGF8 ligands in the extraembryonic ectoderm. Fine chimeric studies with single-cell resolution and expression studies with specific inhibitors for HS movement demonstrated that proteolytic cleavage of HS chains can spread FGF signaling to adjacent cells within a short distance. Together, the results show that spatiotemporal expression of cell surface-tethered HS chains regulate the local reception of FGF-signaling activity during mammalian embryogenesis.