title: Epigenetic switching by the metabolism-sensing factors in the generation of orexin neurons from mouse embryonic stem cells.
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.
authors.affiliation: Laboratory of Cellular Biochemistry, Department of Animal Resource Sciences/Veterinary Medical Sciences, The University of Tokyo, Tokyo 113-8657, Japan.
authors.forename: Koji
authors.initials: K
authors.lastname: Hayakawa
authors.forename: Mitsuko
authors.initials: M
authors.lastname: Hirosawa
authors.forename: Yasuyuki
authors.initials: Y
authors.lastname: Tabei
authors.forename: Daisuke
authors.initials: D
authors.lastname: Arai
authors.forename: Satoshi
authors.initials: S
authors.lastname: Tanaka
authors.forename: Noboru
authors.initials: N
authors.lastname: Murakami
authors.forename: Shintaro
authors.initials: S
authors.lastname: Yagi
authors.forename: Kunio
authors.initials: K
authors.lastname: Shiota
mesh_terms: D000107:Acetylation; D000818:Animals; D002454:Cell Differentiation; D003562:Cytidine / Q000031:analogs & derivatives / Q000494:pharmacology; D019175:DNA Methylation; D015257:DNA-Cytosine Methylases / Q000037:antagonists & inhibitors / Q000378:metabolism; D003841:Deoxycytidine / Q000494:pharmacology; D053595:Embryonic Stem Cells / Q000187:drug effects / Q000502:physiology*; D044127:Epigenesis, Genetic / Q000502:physiology*; D005260:Female; D006031:Glycosylation; D006595:Hexosamines / Q000494:pharmacology; D056572:Histone Deacetylase Inhibitors / Q000494:pharmacology; D006657:Histones / Q000378:metabolism; D006877:Hydroxamic Acids / Q000494:pharmacology; D047908:Intracellular Signaling Peptides and Proteins / Q000235:genetics / Q000378:metabolism*; D051379:Mice; D008810:Mice, Inbred C57BL; D009474:Neurons / Q000378:metabolism*; D009479:Neuropeptides / Q000235:genetics / Q000378:metabolism*; D000068797:Orexins; D011499:Protein Processing, Post-Translational; D017422:Sequence Analysis, DNA; D056564:Sirtuin 1 / Q000037:antagonists & inhibitors / Q000378:metabolism; D018874:Spheroids, Cellular; D024363:Transcription Initiation Site
chemical_list: D006595:Hexosamines; D056572:Histone Deacetylase Inhibitors; D006657:Histones; D006877:Hydroxamic Acids; D047908:Intracellular Signaling Peptides and Proteins; D009479:Neuropeptides; D000068797:Orexins; D003841:Deoxycytidine; C012589:trichostatin A; D003562:Cytidine; C009131:pyrimidin-2-one beta-ribofuranoside; D015257:DNA-Cytosine Methylases; C470545:Sirt1 protein, mouse; D056564:Sirtuin 1; C002022:N-acetylmannosamine
keywords: DNA Methylation; Embryonic Stem Cell; Epigenetics; Histone Acetylase; Histone Acetylation; Mgea5; Neurogenesis; O-GlcNAcylation; Ogt; Sirt1