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SRX8047025: GSM4451165: JD-23; Taeniopygia guttata; RNA-Seq
1 ILLUMINA (NextSeq 500) run: 37.2M spots, 2.8G bases, 1Gb downloads

Submitted by: NCBI (GEO)
Study: DNA methylation regulates transcription factor specific neurodevelopmental but not sexually dimorphic gene expression dynamics in zebra finch telencephalon [RNA-Seq]
show Abstracthide Abstract
Song learning in zebra finches is a prototypical example of a complex learned behavior, yet knowledge on the underlying molecular processes is limited. Therefore, we characterized transcriptomic (RNA sequencing) and epigenomic (RRBS, reduced representation bisulfite sequencing; immunofluorescence) dynamics in matched zebra finch telencephalon samples of both sexes from 1 day post hatching (1 dph) to adulthood, spanning the critical period for song learning (20 dph and 65 dph). We identified extensive transcriptional neurodevelopmental changes during postnatal telencephalon development. DNA (hydroxy)methylation was very low, yet increased over time, particularly in song control nuclei. Only a small fraction of the massive differential expression in the developing zebra finch telencephalon could be explained by differential CpG and CpH DNA methylation. However, a strong association between DNA methylation and age dependent gene expression was found for various transcription factors (i.a. OTX2, AR and FOS) involved in neurodevelopment. Additionally, genomic regions featured by age dependent differential methylation in differentially expressed genes were significantly enriched for specific transcription factor binding motifs. Incomplete dosage compensation was found to be largely responsible for sexually dimorphic gene expression, with dosage compensation increasing throughout life. In conclusion, our results indicate that DNA methylation regulates neurodevelopmental gene expression dynamics through steering transcription factor activity, but does not explain sexually dimorphic gene expression patterns in zebra finch telencephalon. Overall design: For each sex per time point (1 day post hatch (dph), 20dph, 65dph and adult), 3 independent biological replicates were sequenced, resulting in a total of 24 samples. To increase power, each of these biological replicates consisted of a pool of RNA of 3 telencephalons in equal proportions.
Sample: JD-23
SAMN14525593 • SRS6418464 • All experiments • All runs
Library:
Instrument: NextSeq 500
Strategy: RNA-Seq
Source: TRANSCRIPTOMIC
Selection: cDNA
Layout: SINGLE
Construction protocol: Brains from male and female juvenile and adult zebra finches (age: 1, 20, 65 (± 3 days) and adults (> 170) dph) were used, with birds from one nest randomly assigned to different age groups. Birds were rapidly decapitated, and the brain immediately removed and handled under ribonuclease-free conditions. The telencephalon was separated from the rest of the brain. The brain tissue was rapidly frozen on dry ice and stored at -80°C until further processing. Telencephalon samples were lysed in QiazolTM lysis reagent (Qiagen 79306) using a TissueLyser instrument (Qiagen 85300). RNA was extracted using the RNeasy mini kit (Qiagen 74106), according to the manufacturer's instructions. Quality control and library preparation for RNA sequencing was performed by the NXTGNT sequencing facility of Ghent University (Ghent, Belgium). RNA samples were quantified, and quality controlled using the Quant-iTTM Ribogreen® RNA Assay (Invitrogen R11491) kit and Agilent RNA 6000 Nano (Agilent 5067-1511; on Agilent Bioanalyzer). Upon DNase treatment, library preparation (Truseq stranded mRNA, Illumina RS-122-2101 and RS-122-2102) was performed on 1µg of each sample. As recommended by Illumina, fragmentation was performed for 8 minutes at 94°C. Samples were pooled and subsequently multiplex identifiers were added to each pool. To increase uniformity of the protocol, library preparation was performed using the IP-star® Compact Automated System (Diagenode B03000002). Bioanalyzer QC after library preparation (Agilent, High Sensitivity DNA kit) demonstrated no relevant batch effects. Single-end 75 bp sequencing was performed three times for each of the pooled libraries on the Illumina NextSeq500 (over four lanes).
Experiment attributes:
GEO Accession: GSM4451165
Links:
Runs: 1 run, 37.2M spots, 2.8G bases, 1Gb
Run# of Spots# of BasesSizePublished
SRR1147059737,204,7252.8G1Gb2021-04-07

ID:
10488920

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