The Notch response inhibitor DAPT enhances neuronal differentiation in embryonic stem cell‐derived embryoid bodies independently of sonic hedgehog signaling

Crawford, Timothy Q.; Roelink, Henk

doi:10.1002/dvdy.21083

Cited by 90 publications

(85 citation statements)

References 67 publications

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“…S4), since the paucity of supporting cells in the organ of Corti makes reliable transcription factor ChIP currently impractical. NPCs, like supporting cells, rely on active Notch signaling for maintenance of their progenitor state (Ohtsuka et al, 2001;Hatakeyama et al, 2004;Crawford and Roelink, 2007;Borghese et al, 2010). Consistent with this, we observed more than 5-fold (P<0.005) upregulation of Atoh1 and a complete loss of Hes5 mRNA in NPCs following DAPT treatment (Fig.…”

Section: Atoh1 Is Upregulated In Prosensory Progenitors Not Just In supporting

confidence: 86%

Selection of cell fate in the organ of Corti involves the integration of Hes/Hey signaling at the Atoh1 promoter

2016

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Determination of cell fate within the prosensory domain of the developing cochlear duct relies on the temporal and spatial regulation of the bHLH transcription factor Atoh1. Auditory hair cells and supporting cells arise in a wave of differentiation that patterns them into discrete rows mediated by Notch-dependent lateral inhibition. However, the mechanism responsible for selecting sensory cells from within the prosensory competence domain remains poorly understood. We show in mice that rather than being upregulated in rows of cells, Atoh1 is subject to transcriptional activation in groups of prosensory cells, and that highly conserved sites for Hes/Hey repressor binding in the Atoh1 promoter are needed to select the hair cell and supporting cell fate. During perinatal supporting cell transdifferentiation, which is a model of hair cell regeneration, we show that derepression is sufficient to induce Atoh1 expression, suggesting a mechanism for priming the 3′ Atoh1 autoregulatory enhancer needed for hair cell expression.

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Section: Atoh1 Is Upregulated In Prosensory Progenitors Not Just In supporting

confidence: 86%

Selection of cell fate in the organ of Corti involves the integration of Hes/Hey signaling at the Atoh1 promoter

2016

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“…32 Finally, DAPT supplementation significantly reduces notch-dependant transcription within PEG hydrogels, and efficiently prevents glial differentiation within both PEG hydrogel and monolayer cultures. This result is consistent with previous studies showing that constitutive notch activation promotes gliogenesis in peripheral and central nervous system progenitors 62 and that notch inhibition induces a severe neurogenic phenotype in zebrafish, 37 increases neuronal differentiation in neuralized embryoid bodies, 38 and accelerates neuronal differentiation in monolayer cultures of human embryonic stem cell-derived neural stem cells. 10 Interruption of notch signaling is thought to be the mechanism by which DAPT supplementation prevents glial differentiation; however, further mechanistic studies will be needed to confirm that other g-secretase substrates such as E-cadherin and ErbB-4 do not also play a role.…”

Section: Cell Composition In the Presence Of Bfgf Egf And Daptsupporting

confidence: 93%

“…DAPT supplementation may be an effective approach given that, even in cell dense cultures, DAPT can inhibit the ligand-dependent, g-secretase-mediated cleavage and activation of notch receptors that is required for the onset of gliogenesis. 10,11,37,38 Materials and Methods…”

Section: Introductionmentioning

confidence: 99%

Control of Neural Cell Composition in Poly(Ethylene Glycol) Hydrogel Culture with Soluble Factors

Mooney

Haeger

Lawal

et al. 2011

Tissue Engineering Part A

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Poly(ethylene glycol) (PEG) hydrogels are being developed as cell delivery vehicles that have great potential to improve neuronal replacement therapies. Current research priorities include (1) characterizing neural cell growth within PEG hydrogels relative to standard culture systems and (2) generating neuronal-enriched populations within the PEG hydrogel environment. This study compares the percentage of neural precursor cells (NPCs), neurons, and glia present when dissociated neural cells are seeded within PEG hydrogels relative to standard monolayer culture. Results demonstrate that PEG hydrogels enriched the initial cell population for NPCs, which subsequently gave rise to neurons, then to glia. Relative to monolayer culture, PEG hydrogels maintained an increased percentage of NPCs and a decreased percentage of glia. This neurogenic advantage of PEG hydrogels is accentuated in the presence of basic fibroblast growth factor and epidermal growth factor, which more potently increase NPC and neuronal expression markers when applied to cells cultured within PEG hydrogels. Finally, this work demonstrates that glial differentiation can be selectively eliminated upon supplementation with a g-secretase inhibitor. Together, this study furthers our understanding of how the PEG hydrogel environment influences neural cell composition and also describes select soluble factors that are useful in generating neuronal-enriched populations within the PEG hydrogel environment.

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“…Two different approaches were undertaken: a pharmacological treatment with the ␥-secretase inhibitor DAPT, and Notch1 gene silencing. DAPT is a well known Notch1 pharmacological inhibitor (Sastre et al, 2001;Geling et al, 2002;Crawford and Roelink, 2007), preventing Notch1 intracellular cleavage, which is required for NICD nuclear translocation and signaling activation. DAPT exerts a strong inhibitory effect on Notch1 activation, but it has poor selectivity, because of the high number of ␥-secretase substrates (Lleó , 2008).…”

Section: Discussionmentioning

confidence: 99%

Nuclear Factor κB-Dependent Neurite Remodeling Is Mediated by Notch Pathway

Bonini¹,

Ferrari-Toninelli²,

Uberti³

et al. 2011

J. Neurosci.

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In this study, we evaluated whether a cross talk between nuclear factor B (NF-B) and Notch may take place and contribute to regulate cell morphology and/or neuronal network in primary cortical neurons. We found that lack of p50, either induced acutely by inhibiting p50 nuclear translocation or genetically in p50 Ϫ/Ϫ mice, results in cortical neurons characterized by reduced neurite branching, loss of varicosities, and Notch1 signaling hyperactivation. The neuronal morphological effects found in p50 Ϫ/Ϫ cortical cells were reversed after treatment with the ␥-secretase inhibitor DAPT (N-[N-(3,5-difluorophenacetyl)-1-alanyl 1]-S-phenylglycine t-butyl ester) or Notch RNA interference. Together, these data suggested that morphological abnormalities in p50 Ϫ/Ϫ cortical neurons were dependent on Notch pathway hyperactivation, with Notch ligand Jagged1 being a major player in mediating such effect. In this line, we demonstrated that the p50 subunit acts as transcriptional repressor of Jagged1. We also found altered distribution of Notch1 and Jagged1 immunoreactivity in the cortex of p50 Ϫ/Ϫ mice compared with wild-type littermates at postnatal day 1. These data suggest the relevance of future studies on the role of Notch/NF-B cross talk in regulating cortex structural plasticity in physiological and pathological conditions.

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The Notch response inhibitor DAPT enhances neuronal differentiation in embryonic stem cell‐derived embryoid bodies independently of sonic hedgehog signaling

Cited by 90 publications

References 67 publications

Selection of cell fate in the organ of Corti involves the integration of Hes/Hey signaling at the Atoh1 promoter

Selection of cell fate in the organ of Corti involves the integration of Hes/Hey signaling at the Atoh1 promoter

Control of Neural Cell Composition in Poly(Ethylene Glycol) Hydrogel Culture with Soluble Factors

Nuclear Factor κB-Dependent Neurite Remodeling Is Mediated by Notch Pathway

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