RRC ID 44415
Author Hayakawa K, Hirosawa M, Tabei Y, Arai D, Tanaka S, Murakami N, Yagi S, Shiota K.
Title Epigenetic switching by the metabolism-sensing factors in the generation of orexin neurons from mouse embryonic stem cells.
Journal J. Biol. Chem.
Abstract The orexin system plays a central role in the integration of sleep/wake and feeding behaviors in a broad spectrum of neural-metabolic physiology. Orexin-A and orexin-B are produced by the cleavage of prepro-orexin, which is encoded on the Hcrt gene. To date, methods for generating other peptide neurons could not induce orexin neurons from pluripotent stem cells. Considering that the metabolic status affects orexin expression, we supplemented the culture medium with a nutrient factor, ManNAc, and succeeded in generating functional orexin neurons from mouse ES cells. Because DNA methylation inhibitors and histone deacetylase inhibitors could induce Hcrt expression in mouse ES cells, the epigenetic mechanism may be involved in this orexin neurogenesis. DNA methylation analysis showed the presence of a tissue-dependent differentially methylated region (T-DMR) around the transcription start site of the Hcrt gene. In the orexin neurons induced by supplementation of ManNAc, the T-DMR of the Hcrt gene was hypomethylated in association with higher H3/H4 acetylation. Concomitantly, the histone acetyltransferases p300, CREB-binding protein (CBP), and Mgea5 (also called O-GlcNAcase) were localized to the T-DMR in the orexin neurons. In non-orexin-expressing cells, H3/H4 hypoacetylation and hyper-O-GlcNAc modification were observed at the T-DMRs occupied by O-GlcNAc transferase and Sirt1. Therefore, the results of the present study suggest that the glucose metabolite, ManNAc, induces switching from the inactive state by Ogt-Sirt1 to the active state by Mgea5, p300, and CBP at the Hcrt gene locus.
Volume 288(24)
Pages 17099-110
Published 2013-6-14
DOI 10.1074/jbc.M113.455899
PII M113.455899
PMID 23625921
PMC PMC3682516
MeSH Acetylation Animals Cell Differentiation Cytidine / analogs & derivatives Cytidine / pharmacology DNA Methylation DNA-Cytosine Methylases / antagonists & inhibitors DNA-Cytosine Methylases / metabolism Deoxycytidine / pharmacology Embryonic Stem Cells / drug effects Embryonic Stem Cells / physiology* Epigenesis, Genetic / physiology* Female Glycosylation Hexosamines / pharmacology Histone Deacetylase Inhibitors / pharmacology Histones / metabolism Hydroxamic Acids / pharmacology Intracellular Signaling Peptides and Proteins / genetics Intracellular Signaling Peptides and Proteins / metabolism* Mice Mice, Inbred C57BL Neurons / metabolism* Neuropeptides / genetics Neuropeptides / metabolism* Orexins Protein Processing, Post-Translational Sequence Analysis, DNA Sirtuin 1 / antagonists & inhibitors Sirtuin 1 / metabolism Spheroids, Cellular Transcription Initiation Site
IF 4.011
Times Cited 22
WOS Category BIOCHEMISTRY & MOLECULAR BIOLOGY
Resource
Human and Animal Cells