RRC ID 56066
Author Méndez-Maldonado K, Vega-López G, Caballero-Chacón S, Aybar MJ, Velasco I.
Title Activation of Hes1 and Msx1 in Transgenic Mouse Embryonic Stem Cells Increases Differentiation into Neural Crest Derivatives.
Journal Int J Mol Sci
Abstract The neural crest (NC) comprises a multipotent cell population that produces peripheral neurons, cartilage, and smooth muscle cells, among other phenotypes. The participation of Hes1 and Msx1 when expressed in mouse embryonic stem cells (mESCs) undergoing NC differentiation is unexplored. In this work, we generated stable mESCs transfected with constructs encoding chimeric proteins in which the ligand binding domain of glucocorticoid receptor (GR), which is translocated to the nucleus by dexamethasone addition, is fused to either Hes1 (HGR) or Msx1 (MGR), as well as double-transgenic cells (HGR+MGR). These lines continued to express pluripotency markers. Upon NC differentiation, all lines exhibited significantly decreased Sox2 expression and upregulated Sox9, Snai1, and Msx1 expression, indicating NC commitment. Dexamethasone was added to induce nuclear translocation of the chimeric proteins. We found that Collagen IIa transcripts were increased in MGR cells, whereas coactivation of HGR+MGR caused a significant increase in Smooth muscle actin (α-Sma) transcripts. Immunostaining showed that activation in HGR+MGR cells induced higher proportions of β-TUBULIN III⁺, α-SMA⁺ and COL2A1⁺ cells. These findings indicate that nuclear translocation of MSX-1, alone or in combination with HES-1, produce chondrocyte-like cells, and simultaneous activation of HES-1 and MSX-1 increases the generation of smooth muscle and neuronal cells.
Volume 19(12)
Published 2018-12-13
DOI 10.3390/ijms19124025
PII ijms19124025
PMID 30551562
PMC PMC6321090
MeSH Actins / genetics Animals Cell Differentiation Cell Nucleus / metabolism Cells, Cultured Chondrocytes / cytology* Chondrocytes / metabolism Collagen Type II / genetics Dexamethasone / pharmacology MSX1 Transcription Factor / genetics* MSX1 Transcription Factor / metabolism Mice Mice, Transgenic Mouse Embryonic Stem Cells / cytology* Mouse Embryonic Stem Cells / metabolism Myocytes, Smooth Muscle / cytology* Myocytes, Smooth Muscle / metabolism NIH 3T3 Cells Neural Crest / cytology* Neural Crest / metabolism Promoter Regions, Genetic Protein Transport / drug effects Receptors, Glucocorticoid / genetics* Recombinant Fusion Proteins / metabolism SOX9 Transcription Factor / metabolism SOXB1 Transcription Factors / metabolism Snail Family Transcription Factors / metabolism Transcription Factor HES-1 / genetics* Transcription Factor HES-1 / metabolism
IF 4.183
Times Cited 1
Human and Animal Cells MC3T3-G2/PA6(RCB1127)