Morphine delays neural stem cells differentiation by facilitating Nestin overexpression
| Autor: | Adrián García-Concejo, Fernando León-Lobera, Ada Jimenez-Gonzalez, Raquel E. Rodríguez |
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| Rok vydání: | 2017 |
| Předmět: |
0301 basic medicine
Homeobox protein NANOG Embryo Nonmammalian Neurogenesis Biophysics Transcription factor complex macromolecular substances Biology Biochemistry Animals Genetically Modified Histones Nestin 03 medical and health sciences Mice SOX2 Neural Stem Cells Genes Reporter Animals Humans Molecular Biology reproductive and urinary physiology SOX Transcription Factors Binding Sites Morphine Naloxone fungi Nanog Homeobox Protein Gene Expression Regulation Developmental Acetylation DNA Methylation Zebrafish Proteins Molecular biology Neural stem cell Up-Regulation 030104 developmental biology nervous system embryonic structures DNA methylation CpG Islands Stem cell E1A-Associated p300 Protein Octamer Transcription Factor-3 Protein Processing Post-Translational |
| Zdroj: | Biochimica et biophysica acta. General subjects. 1862(3) |
| ISSN: | 0304-4165 |
| Popis: | Background Morphine is used as an analgesic although it causes important secondary effects. These effects are triggered by several mechanisms leading to the dysregulation of gene expression. Here we aimed to study these alterations on neural stem cells (NSC) during CNS development. Methods AB strain and tg nestin:GFP zebrafish embryos, zebrafish primary neuron culture and mouse embryonic stem cells were used to assess the effect of morphine by qPCR, time lapse microscopy and western blot. ChIP-qPCR and bisulfite conversion assay were performed to determine the changes exerted by morphine in a Nestin candidate enhancer. Results Morphine increases GFP in nestin:GFP embryos and overexpresses the NSC marker Nestin. Morphine also exerts a hyperacetylation effect on H3K27 and decreases DNA methylation within a region located 18 Kb upstream nestin transcription starting site. Here, a binding site for the transcription factor complex Sox2/Oct4/Nanog was predicted. These factors are also upregulated by morphine. Besides, morphine increases the histone acetyl transferase p300. The inhibition of p300 activity decreases Nestin. Conclusions Morphine facilitates Nestin increase by several mechanisms which include hyperacetylation of H3K27, decreased DNA methylation and the overexpression of the transcription factors sox2, oct4 and nanog. It has also been demonstrated that nestin levels depend on p300 activity. The facilitated Nestin expression delays the normal differentiation of neural stem cells. General significance The present work provides novel evidence of the effects induced by morphine in the normal differentiation of NSCs, altering Nestin through changes on p300, H3K27ac, DNA methylation and Oct4, Sox2, and Nanog. |
| Databáze: | OpenAIRE |
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