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  • Neuronal developmental gene and miRNA signatures induced by histone deacetylase inhibitors in human embryonic stem cells.

Neuronal developmental gene and miRNA signatures induced by histone deacetylase inhibitors in human embryonic stem cells.

Cell death & disease (2015-05-08)
K Meganathan, S Jagtap, S P Srinivasan, V Wagh, J Hescheler, J Hengstler, M Leist, A Sachinidis
ABSTRACT

Human embryonic stem cells (hESCs) may be applied to develop human-relevant sensitive in vitro test systems for monitoring developmental toxicants. The aim of this study was to identify potential developmental toxicity mechanisms of the histone deacetylase inhibitors (HDAC) valproic acid (VPA), suberoylanilide hydroxamic acid (SAHA) and trichostatin A (TSA) relevant to the in vivo condition using a hESC model in combination with specific differentiation protocols and genome-wide gene expression and microRNA profiling. Analysis of the gene expression data showed that VPA repressed neural tube and dorsal forebrain (OTX2, ISL1, EMX2 and SOX10)-related transcripts. In addition, VPA upregulates axonogenesis and ventral forebrain-associated genes, such as SLIT1, SEMA3A, DLX2/4 and GAD2. HDACi-induced expression of miR-378 and knockdown of miR-378 increases the expression of OTX2 and EMX2, which supports our hypothesis that HDACi targets forebrain markers through miR-378. In conclusion, multilineage differentiation in vitro test system is very sensitive for monitoring molecular activities relevant to in vivo neuronal developmental toxicity. Moreover, miR-378 seems to repress the expression of the OTX2 and EMX2 and therefore could be a regulator of the development of neural tube and dorsal forebrain neurons.

MATERIALS
Product Number
Brand
Product Description

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Monoclonal Anti-MAP2 antibody produced in mouse, clone HM-2, purified from hybridoma cell culture
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Chloroform, anhydrous, ≥99%, contains 0.5-1.0% ethanol as stabilizer
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Chloroform, anhydrous, contains amylenes as stabilizer, ≥99%
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Chloroform, ≥99%, PCR Reagent, contains amylenes as stabilizer
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Methanol, anhydrous, 99.8%
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Methanol, purification grade, 99.8%
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Methanol, suitable for NMR (reference standard)
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DAPI, for nucleic acid staining
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Anti-β-Tubulin III antibody produced in rabbit, affinity isolated antibody, buffered aqueous solution
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Methanol solution, contains 0.50 % (v/v) triethylamine
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SAHA, ≥98% (HPLC)
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Phenylacetic acid, ≥99%, FCC, FG
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Valproic acid sodium salt, 98%
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Triton X-100, laboratory grade
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Trichostatin A , Vetec, reagent grade, from Streptomyces sp., ≥98%