A role for DNA methylation in gastrulation and somite patterning.
DNA methylation constitutes an important epigenetic factor in the control of genetic information. In this study, we analyzed expression of the DNA methyltransferase gene and examined DNA methylation patterns during early development of the zebrafish. Maternal transcripts of the zebrafish DNA methyltransferase gene (MTase) are ubiquitously present at high levels in early embryos with overall levels decreasing after the blastula stage. At 24 h, methyltransferase mRNA is predominantly found in the brain, neural tube, eyes, and differentiating somites. Expression of MTase in the somites is highest in the anterior cells of the somites. Despite the high levels of MTase mRNA in blastula-stage embryos, we observe DNA hypomethylation at the blastula and gastrula stages compared to sperm or older embryos. Zebrafish embryos treated with 5-azacytidine (5-azaC) and 5-aza-2-deoxycytidine (5-azadC), nucleotide analogs known to induce cellular differentiation and DNA hypomethylation in mammalian cells, exhibit DNA hypomethylation and developmental perturbations. These defects are specifically observed in embryos treated at the beginning of the blastula period, just prior to midblastula transition. The most common phenotype is the loss of tail and abnormal patterning of somites. Head development is also affected in some embryos. Histological and in situ hybridization analyses reveal whole or partial loss of a differentiated notochord and midline muscle in treated embryos. When examined during gastrulation, 5-azaC-treated embryos have a shortened and thickened axial mesoderm. We propose that DNA methylation is required for normal gastrulation and subsequent patterning of the dorsal mesoderm.
Martin
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CC
Department of Cellular and Molecular Medicine, Loeb Health Research Institute, Ottawa Hospital, University of Ottawa, 725 Parkdale Avenue, Ottawa, Ontario, K1Y 4E9, Canada.
Laforest
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L
Akimenko
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MA
Ekker
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M
D000014:Abnormalities, Drug-Induced / Q000196:embryology / Q000235:genetics; D000595:Amino Acid Sequence; D000818:Animals; D001374:Azacitidine / Q000031:analogs & derivatives / Q000633:toxicity; D019521:Body Patterning / Q000187:drug effects / Q000235:genetics / Q000502:physiology*; D019175:DNA Methylation / Q000187:drug effects; D000077209:Decitabine; D005775:Gastrula / Q000378:metabolism*; D018507:Gene Expression Regulation, Developmental; D015971:Gene Expression Regulation, Enzymologic; D006801:Humans; D008780:Methyltransferases / Q000235:genetics; D008969:Molecular Sequence Data; D009132:Muscles / Q000002:abnormalities / Q000187:drug effects / Q000196:embryology; D009672:Notochord / Q000002:abnormalities / Q000187:drug effects / Q000196:embryology; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D017386:Sequence Homology, Amino Acid; D019170:Somites / Q000378:metabolism*; D015027:Zebrafish / Q000196:embryology* / Q000235:genetics / Q000378:metabolism*