Activation of Hes1 and Msx1 in Transgenic Mouse Embryonic Stem Cells Increases Differentiation into Neural Crest Derivatives

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.

Neural Stem Cell-Derived Exosomes Regulate Neural Stem Cell Differentiation Through miR-9-Hes1 Axis

“…Our results are supported by others' studies that investigate the effects of Hes1 on human NSC differentiation (Yang et al, 2020). Thus, our finding, together with others' observations (Kobayashi and Kageyama, 2010;Mendez-Maldonado et al, 2018;Yang et al, 2020), indicates that the involvement of Notch signaling and Hes1 in NSC regulation is highly time-and condition-dependent. During CNS development, Hes1 acts as a key neural fate determinant in early stage and then functions as an anti-neural regulator in post-natal stage (Kobayashi and Kageyama, 2010;Mendez-Maldonado et al, 2018).…”

“…Thus, our finding, together with others’ observations ( Kobayashi and Kageyama, 2010 ; Mendez-Maldonado et al, 2018 ; Yang et al, 2020 ), indicates that the involvement of Notch signaling and Hes1 in NSC regulation is highly time- and condition-dependent. During CNS development, Hes1 acts as a key neural fate determinant in early stage and then functions as an anti-neural regulator in post-natal stage ( Kobayashi and Kageyama, 2010 ; Mendez-Maldonado et al, 2018 ). Furthermore, Hes1 has been reported to negatively regulate Notch signaling in a feedback manner during CNS development ( Kobayashi and Kageyama, 2010 ; Boareto et al, 2017 ).…”

Nasal Septum Deviation as the Consequence of BMP-Controlled Changes to Cartilage Properties

“…Overall, canonical WNT signaling apparent by nuclear NPBC staining was increased in the mutant septum, particularly at the site of deviation, whereas cytoplasmic NPBC staining was observed in BMP7 ctrl mice. Loss of BMP7 was associated with a reduction in MSX1, a gene downstream of BMP signaling that promotes chondrogenic differentiation of NCC ( Méndez-Maldonado et al, 2018 ), at both P14 ( p < 0.05) and P30 ( p < 0.001) timepoints ( Figure 5C ). Gremlin 1 (GREM1), a BMP antagonist and inhibitor of chondrocyte hypertrophy ( Zhong et al, 2016a ), was significantly decreased once septum deviation was established.…”

The crucial role of model systems in understanding the complexity of cell signaling in human neurocristopathies

“…Several studies have generated NCCs in vitro with a variety of approaches that include stromal‐derived inducing activity for neural commitment and subsequent stimulation with different concentrations of BMP4 to obtain peripheral neurons, glia, and smooth muscle cells from the NC, starting with mouse or primate ESCs (Mizuseki et al, 2003). In recent work, we observed increases in Tuj1+ neurons, smooth muscle cells expressing smooth muscle actin ( sma ), and chondrocytes positive for col2a1 when transgenic mouse ESCs overexpressing Msx1 and Hes1 were differentiated to NCC/NCCs with the stromal cell inducing activity method (Méndez‐Maldonado et al, 2018). A second method to obtain NCCs from mouse ESCs is through differentiation via embryoid body (EB) formation.…”