The transcription factor Pou3f1 promotes neural fate commitment via activation of neural lineage genes and inhibition of external signaling pathways

Abstract

The neural fate commitment of pluripotent stem cells requires the repression of extrinsic inhibitory signals and the activation of intrinsic positive transcription factors. However, how these two events are integrated to ensure appropriate neural conversion remains unclear. In this study, we showed that Pou3f1 is essential for the neural differentiation of mouse embryonic stem cells (ESCs), specifically during the transition from epiblast stem cells (EpiSCs) to neural progenitor cells (NPCs). Chimeric analysis showed that Pou3f1 knockdown leads to a markedly decreased incorporation of ESCs in the neuroectoderm. By contrast, Pou3f1-overexpressing ESC derivatives preferentially contribute to the neuroectoderm. Genome-wide ChIP-seq and RNA-seq analyses indicated that Pou3f1 is an upstream activator of neural lineage genes, and also is a repressor of BMP and Wnt signaling. Our results established that Pou3f1 promotes the neural fate commitment of pluripotent stem cells through a dual role, activating internal neural induction programs and antagonizing extrinsic neural inhibitory signals.DOI: http://dx.doi.org/10.7554/eLife.02224.001

Dynamic Heterogeneity of Brachyury in Mouse Epiblast Stem Cells Mediates Distinct Response to Extrinsic Bone Morphogenetic Protein (BMP) Signaling

“…Whole-mount in Situ Hybridization-Whole-mount in situ hybridizations were performed as described previously (69). The probes for Oct4, Nanog, and Brachyury were PCR-amplified from mouse cDNA.…”

Is this a brain which I see before me? Modeling human neural development with pluripotent stem cells

“…Similarly, Smad-interacting protein 1 (Sip1; also known as Zeb2) (Chng et al, 2010) and Smad7 (Ozair et al, 2013) were found to act as key intrinsic neural inducers from PSCs, by directly inhibiting the downstream effectors of BMP and Activin signaling. Oct6 (also known as Pou3f1) is another positive inducer of neural fate identified in PSC models, which acts through repression of downstream targets of BMP and Wnt signaling (Zhu et al, 2014).…”

Histone deacetylation promotes mouse neural induction by restricting Nodal-dependent mesendoderm fate

“…Chimeric embryos analysis was performed as described previously 45 . Briefly, GFP-labelled WT mESCs (HDAC1 lox/lox ) and HDAC1 KO mESCs (HDAC1 D/D ) were injected into E2.5 mouse blastocysts, respectively, and then transferred into the uteri of day 2.5 pseudopregnant female mice.…”

“…For gene knockdown in mESCs, the corresponding short hairpin RNAs (shRNAs) and control shRNA against Luciferase gene were cloned into the pLKO.1 constructs (Addgene). For the overexpression of HDAC1, the complementary DNA was inserted into pCDH-EF1-Puro vector (Systems Biosciences), the production and infection of lentivirus supernatant was performed as described previously 45 . Briefly, after infection of lentivirus for 24 h, stable mESCs were selected by puromycin (2 mg ml À 1 , Sigma).…”

“…To this, GFP-labelled wild-type (WT) or HDAC1 knockout (KO) mESCs were injected into the blastocyst, and embryos were transferred into pseudopregnant mice (see methods for details) 45 . Chimeric embryos were collected at E8.5-E9.0, and the contribution of GFP þ cells was quantified in the chimeras generated from WT or HDAC1 KO mESCs, respectively.…”