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SRX273741: GSM1133393: 16-cell.BSseq; Danio rerio; Bisulfite-Seq
2 ILLUMINA (Illumina HiSeq 2000) runs: 368.9M spots, 74.5G bases, 49.7Gb downloads

Submitted by: Gene Expression Omnibus (GEO)
Study: Sperm but not oocyte DNA methylome is inherited by zebrafish early embyros
show Abstracthide Abstract
5-methylcytosine is a major epigenetic modification sometimes called "the fifth nucleotide". However, our knowledge of how offspring inherit the DNA methylome from parents is limited. We generated nine single-base resolution DNA methylomes including zebrafish gametes and early embryos. The oocyte methylome is significantly hypo-methylated compared to sperm. Strikingly, the paternal DNA methylation pattern is maintained throughout early embryogenesis. The maternal DNA methylation pattern is maintained until the 16-cell stage. Then, the oocyte methylome is gradually discarded through cell division, and progressively reprogrammed to a pattern similar to that of the sperm methylome. The passive demethylation rate and the de novo methylation rate are similar in the maternal DNA. By the midblastula stage, the embryo?s methylome is virtually identical to the sperm methylome. Moreover, inheritance of the sperm methylome facilitates the epigenetic regulation of embryogenesis. Therefore, besides DNA sequences, sperm DNA methylome is also inherited in zebrafish early embryos. Overall design: hMeDIP-seq is performed in sperm,2-cell,16-cell,1k-cell and a input control sample 4 new samples are added to GSE44075. RNA-seq is performed in sperm, egg,1k-cell, germring samples .hMeDIP-seq is performed in sperm,2-cell,16-cell,1k-cell and a input control sample 10 new samples are added to GSE44075. Bisulfite-seq is performed for nine samples : sperm, egg,16-cell,32-cell,64-cell,128-cell,1k-cell , Germring and testis. TAB-seq is performed for one sample , 32-cell.
Sample: 16-cell.BSseq
SAMN02117045 • SRS418158 • All experiments • All runs
Organism: Danio rerio
Library:
Instrument: Illumina HiSeq 2000
Strategy: Bisulfite-Seq
Source: GENOMIC
Selection: RANDOM
Layout: PAIRED
Construction protocol: MethylC-Seq:Genomic DNA (>100ng) spiked with 0.5% unmethylated cl857 Sam7 Lambda DNA (Promega) was sonicated into 100-500bp fragments with Covaris S2 (Applied Biosystems). Fragmented DNA was then subjected to end repair with the end repair enzyme mix (NEB), dA tailing with the Klenow 3´-5´ exo- (NEB) and ligation of methylated multiplexing adapters (Illumina) using T4 DNA ligase (NEB). Adapter-ligated DNA of 280-500bp was purified by 2%agarose gel electrophoresis, and bisulfite conversion was performed using the EZ DNA methylation-Gold kit (Zymo Research) according to the instruction manual. Sperms were released by gently but repeatedly disrupting the testis with a pipette tip in HBSS. Unfertilized oocytes were collected by squeezing the abdomen of females about 5 minutes after spawning. Embryos were grown in embryo medium at 28°C, staged according to standard morphological criteria..Zebrafish embryos were chilled on ice, washed by pre-chilled PBS buffer three times to avoid the contamination and homogenized in 10 vol buffer N (10 mM HEPES, pH 7.5, 250 mM sucrose, 50 mMNaCl, 5 mM MgCl2, 10 µg/ml cytochalasin B, 1 mMdithiothreitol and protease inhibitors) with ten strokes of a loose fitting pestle in a glass homogenizer, and lysates were decanted for 20 min on ice. Then lysates were filtered with 70µm cell strainer(BD Biosciences) and centrifuged through 1 M sucrose at 1000 g for 30 min. Pelleted nuclei from embryos were subjected to genomic DNA extraction by standard phenol precipitation method.
Experiment attributes:
GEO Accession: GSM1133393
Links:
External link:
Runs: 2 runs, 368.9M spots, 74.5G bases, 49.7Gb
Run# of Spots# of BasesSizePublished
SRR837878200,176,06540.4G27.7Gb2013-05-10
SRR837879168,697,24634.1G22.1Gb2013-05-10

ID:
388470

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