Morphine delays neural stem cells differentiation by facilitating Nestin overexpression

• Published in: Biochimica et biophysica acta. General subjects Vol. 1862; no. 3; pp. 474 - 484
• Main Authors: Jimenez-Gonzalez, Ada, García-Concejo, Adrián, León-Lobera, Fernando, Rodriguez, Raquel E.
• Format: Journal Article
• Language: English
• Published: Netherlands Elsevier B.V 01.03.2018
• Subjects: Acetylation - drug effects; Animals; Animals, Genetically Modified; Binding Sites; Central nervous system; CpG Islands - drug effects; Development; DNA Methylation - drug effects; E1A-Associated p300 Protein - physiology; Embryo, Nonmammalian - drug effects; Epigenetics; Gene Expression Regulation, Developmental - drug effects; Genes, Reporter; Histones - metabolism; Humans; Mice; Morphine; Morphine - pharmacology; Naloxone - pharmacology; Nanog Homeobox Protein - biosynthesis; Nanog Homeobox Protein - genetics; Nestin - biosynthesis; Nestin - genetics; Neural Stem Cells - drug effects; Neural Stem Cells - metabolism; Neurogenesis - drug effects; Octamer Transcription Factor-3 - biosynthesis; Octamer Transcription Factor-3 - genetics; Protein Processing, Post-Translational - drug effects; SOX Transcription Factors - biosynthesis; SOX Transcription Factors - genetics; Stem cell; Up-Regulation - drug effects; Zebrafish Proteins - biosynthesis; Zebrafish Proteins - genetics; Stem cell; Central nervous system; CNS; Morphine; OPRM1; ChIP; ES; HATi; CBP; GFP; Development; Epigenetics; HAT; NSC; HPF
• ISSN: 0304-4165; 1872-8006
• DOI: 10.1016/j.bbagen.2017.10.016

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. 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. 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. 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. 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. [Display omitted] •Morphine increases nestin through several mechanisms delaying NSCs differentiation.•The exposure to morphine upregulates sox2, oct4 and nanog.•nestin overexpression after morphine exposure is p300-dependent.•Morphine induces hyperacetylation and demethylation of a nestin candidate enhancer.