title: MeCP2 dependent heterochromatin reorganization during neural differentiation of a novel Mecp2-deficient embryonic stem cell reporter line.
abstract: The X-linked Mecp2 is a known interpreter of epigenetic information and mutated in Rett syndrome, a complex neurological disease. MeCP2 recruits HDAC complexes to chromatin thereby modulating gene expression and, importantly regulates higher order heterochromatin structure. To address the effects of MeCP2 deficiency on heterochromatin organization during neural differentiation, we developed a versatile model for stem cell in vitro differentiation. Therefore, we modified murine Mecp2 deficient (Mecp2(-/y)) embryonic stem cells to generate cells exhibiting green fluorescent protein expression upon neural differentiation. Subsequently, we quantitatively analyzed heterochromatin organization during neural differentiation in wild type and in Mecp2 deficient cells. We found that MeCP2 protein levels increase significantly during neural differentiation and accumulate at constitutive heterochromatin. Statistical analysis of Mecp2 wild type neurons revealed a significant clustering of heterochromatin per nuclei with progressing differentiation. In contrast we found Mecp2 deficient neurons and astroglia cells to be significantly impaired in heterochromatin reorganization. Our results (i) introduce a new and manageable cellular model to study the molecular effects of Mecp2 deficiency, and (ii) support the view of MeCP2 as a central protein in heterochromatin architecture in maturating cells, possibly involved in stabilizing their differentiated state.
authors.affiliation: Department of Biology, Technische Universität Darmstadt, Darmstadt, Germany.
authors.forename: Bianca
authors.initials: B
authors.lastname: Bertulat
authors.forename: Maria Luigia
authors.initials: ML
authors.lastname: De Bonis
authors.forename: Floriana
authors.initials: F
authors.lastname: Della Ragione
authors.forename: Anne
authors.initials: A
authors.lastname: Lehmkuhl
authors.forename: Manuela
authors.initials: M
authors.lastname: Milden
authors.forename: Christian
authors.initials: C
authors.lastname: Storm
authors.forename: K Laurence
authors.initials: KL
authors.lastname: Jost
authors.forename: Simona
authors.initials: S
authors.lastname: Scala
authors.forename: Brian
authors.initials: B
authors.lastname: Hendrich
authors.forename: Maurizio
authors.initials: M
authors.lastname: D'Esposito
authors.forename: M Cristina
authors.initials: MC
authors.lastname: Cardoso
mesh_terms: D000818:Animals; D002460:Cell Line; D053595:Embryonic Stem Cells / Q000166:cytology* / Q000378:metabolism; D017353:Gene Deletion; D018507:Gene Expression Regulation, Developmental; D006570:Heterochromatin / Q000378:metabolism*; D006801:Humans; D051783:Methyl-CpG-Binding Protein 2 / Q000032:analysis / Q000235:genetics / Q000378:metabolism*; D051379:Mice; D055495:Neurogenesis; D009474:Neurons / Q000166:cytology* / Q000378:metabolism; D015518:Rett Syndrome / Q000235:genetics / Q000378:metabolism
chemical_list: D006570:Heterochromatin; D051783:Methyl-CpG-Binding Protein 2