Transcription-independent heritability of induced histone modifications in the mouse preimplantation embryo

• Published in: PloS one 2009-06, Vol.4 (6), p.e6086
• Main Authors: VerMilyea, Matthew D, O'Neill, Laura P, Turner, Bryan M
• Format: Article
• Language: English
• Subjects: Acetates; Acetic acid; Acetylation; Analysis; Animals; Biomedical research; Blastocyst - cytology; Blastocyst - drug effects; Blastocyst - metabolism; CDX2 protein; Cell Biology/Gene Expression; Cell culture; Cell cycle; Cell fate; Chromatin; Developmental Biology/Developmental Molecular Mechanisms; Developmental Biology/Embryology; Developmental Biology/Molecular Development; DNA binding proteins; DNA methylation; Embryo; Embryonic development; Embryos; Enzymes; Epigenesis, Genetic; Epigenetic inheritance; Epigenetics; Epilepsy; Gene expression; Genes; Genetics and Genomics/Epigenetics; Glyceraldehyde-3-phosphate dehydrogenase; Heritability; Histone deacetylase; Histones; Histones - chemistry; Histones - metabolism; HOXB1 protein; Immunoprecipitation; Inhibitors; Lysine; Methylation; Mice; Mice, Inbred BALB C; Microscopy, Fluorescence - methods; Mitosis; Models, Biological; Models, Genetic; Molecular Biology/Histone Modification; Oct-4 protein; Post-translation; Promoter Regions, Genetic; Quantitative analysis; Stem cells; Time Factors; Transcription; Transcription (Genetics); Transcription, Genetic; Valproic acid; Valproic Acid - pharmacology
• ISSN: 1932-6203; 1932-6203
• DOI: 10.1371/journal.pone.0006086

Enzyme-catalyzed, post-translational modifications of core histones have been implicated in the complex changes in gene expression that drive early mammalian development. However, until recently the small number of cells available from the preimplantation embryo itself has prevented quantitative analysis of histone modifications at key regulator genes. The possible involvement of histone modifications in the embryo's response to extracellular signals, or as determinants of cell fate or lineage progression, remains unclear. Here we describe the use of a recently-developed chromatin immunoprecipitation technique (CChIP) to assay histone modification levels at key regulator genes (Pou5f1, Nanog, Cdx2, Hoxb1, Hoxb9) as mouse embryos progress from 8-cell to blastocyst in culture. Only by the blastocyst stage, when the embryonic (Inner Cell Mass) and extra-embryonic (Trophoblast) lineages are compared, do we see the expected association between histone modifications previously linked to active and silent chromatin, and transcriptional state. To explore responses to an environmental signal, we exposed embryos to the histone deacetylase inhibitor, anti-epileptic and known teratogen valproic acid (VPA), during progression from 8-cell to morula stage. Such treatment increased H4 acetylation and H3 lysine 4 methylation at the promoters of Hoxb1 and Hoxb9, but not the promoters of Pou5f1, Nanog,Cdx2 or the housekeeping gene Gapdh. Despite the absence of detectable Hoxb transcription, these VPA-induced changes were heritable, following removal of the inhibitor, at least until the blastocyst stage. The selective hyperacetylation of Hoxb promoters in response to a histone deacetylase inhibitor, suggests that Hox genes have a higher turnover of histone acetates than other genes in the preimplantation embryo. To explain the heritability, through mitosis, of VPA-induced changes in histone modification at Hoxb promoters, we describe how an epigenetic feed-forward loop, based on cross-talk between H3 acetylation and H3K4 methylation, might generate a persistently increased steady-state level of histone acetylation in response to a transient signal.