Unknown,Transcriptomics,Genomics,Proteomics

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Gonadal DNA methylomes of half-smooth tongue sole Cynoglossus semilaevis

ABSTRACT: Environmental sex determination (ESD) occurs in divergent, phylogenetically unrelated taxa, and in some species co-occurs with genetic sex determination (GSD) mechanisms. Although epigenetic regulation in response to environmental effects has long been proposed to be associated with ESD, a systemic analysis on epigenetic regulation of ESD is still lacking. Using half-smooth tongue sole (Cynoglossus semilaevis) as a model – a marine fish which has both ZW chromosomal GSD and temperature-dependent ESD – we investigated the role of DNA methylation in transition from GSD to ESD by comparing gonadal DNA methylomes of parental females, parental pseudo-males, F1 females, F1 pseudo-males and normal males. To assess the gonadal DNA methylome patterns across different sexual types of tongue sole, we carried out BS-seq on bisulfite converted DNA extracted from adult gonads of parental females, parental pseudo-males, and F1 pseudo-males and females from a cross between a parental pseudo-male and a normal female. We also sampled normal male individuals as a control for the normal male DNA methylation pattern. For each of the five samples, two biological replicates were utilized, with each replicate being pooled by five fish. The phenotype and genotype of each selected fish was identified by the histological analysis and PCR validation using the W chromosome specific marker. DNA were isolated from five pooled gonads of the same replicate, then 5 ?g DNA was used to do the bisulfite conversion and BS-seq. The bisulfite conversion of sample DNA was carried out using a modified NH4HSO3-based protocol (Hayatsu et al. 2006). The paired-end library construction and sequencing were carried out using Illumina HiSeq 2000, according to the manufacturer’s instructions (Illumina). We also mixed 25 ng cl857 Sam7 Lambda DNA in each sample to use as conversion quality control for each library.

ORGANISM(S): Cynoglossus semilaevis

SUBMITTER: Hong Cao

PROVIDER: E-GEOD-41129 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Project description:The parasitoid wasp Nasonia vitripennis is an emerging genetic model for functional analysis of DNA methylation. Here, we characterize genome-wide methylation at a base-pair resolution, and compare these results to gene expression across five developmental stages and to methylation patterns reported in other insects. An accurate assessment of DNA methylation across the genome is accomplished using bisulfite sequencing of adult females from a highly inbred line. One-third of genes show extensive methylation over the gene body, yet methylated DNA is not found in non-coding regions and rarely in transposons. Methylated genes occur in small clusters across the genome. Methylation demarcates exon-intron boundaries, with elevated levels over exons, primarily in the 5’ regions of genes. It is also elevated near the sites of translational initiation and termination, with reduced levels in 5’ and 3’ UTRs. Methylated genes have higher median expression levels and lower expression variation across development stages than non-methylated genes. There is no difference in frequency of differential splicing between methylated and non-methylated genes, and as yet no established role for methylation in regulating alternative splicing in Nasonia. Phylogenetic comparisons indicate that many genes maintain methylation status across long evolutionary time scales. Nasonia methylated genes are more likely to be conserved in insects, but even those that are not conserved show broader expression across development than comparable non-methylated genes. Finally, examination of duplicated genes shows that those paralogs that have lost methylation in the Nasonia lineage following gene duplication evolve more rapidly, show decreased median expression levels, and increased specialization in expression across development. Methylation of Nasonia genes signals constitutive transcription across developmental stages, whereas non-methylated genes show more dynamic developmental expression patterns. We speculate that loss of methylation may result in increased developmental specialization in evolution and acquisition of methylation may lead to broader constitutive expression. Whole-genome bisulfite sequencing of 24-hour adult female Nasonia vitripennis whole body samples using Iilumina GAIIx and HiSeq2000.

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Gonadal DNA methylomes of half-smooth tongue sole Cynoglossus semilaevis

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