E-GEOD-43717 - Transcription profiling by high throughput sequencing of purified Sertoli cells, spermatogonia, spermatocytes, spermatids and spermatozoa from mice

Status
Released on 20 June 2013, last updated on 28 June 2016
Organism
Mus musculus
Samples (5)
Protocols (2)
Description
Understanding the extent of genomic transcription and its functional relevance is a central goal in genomics research. However, detailed genome€�wide investigations of transcriptome complexities in major mammalian organs  and  their  underlying  cellular sources, transcriptional mechanisms, and functional relevance have been scarce. Here we first show, using extensive RNA€�seq data, that transcription of both functional and nonfunctional genomic elements is substantially more widespread in the testis than in other organs across representative mammals. By scrutinizing the transcriptomes of all main testicular cell types in the mouse, we then reveal that meiotic  spermatocytes and especially post€�meiotic round spermatids have remarkably diverse transcriptomes, which explains the high transcriptome  complexity of the testis as a whole. The widespread transcriptional activity in spermatocytes and spermatids encompasses protein €�coding genes and long noncoding RNA genes but also poorly conserved intergenic sequences, suggesting that much of it is not of immediate functional relevance.  Rather, our analyses of genome€�wide epigenetic data show that this prevalent transcription, which apparently promoted the birth of new genes during evolution, results from a highly permissive chromatin state during and after meiosis that may ultimately facilitate the replacement of histones by protamines during late spermatogenesis. To study the cellular source and mechanisms of high transcriptome complexity in the mammalian testis, we generated strand-specific deep coverage RNA€�Seq data for purified sertoli cells, spermatogonia, spermatocytes, spermatids and spermatozoa as well as for brain, liver and the whole testis from the mouse. We prepared 8 sequencing libraries for the polyadenylated RNA fraction of each sample and sequenced each library in 3 lanes of the Illumina Genome Analyser IIx platform, yielding a total of >60 millions strand-specific reads of 76 base pairs per sample. In addition, we generated ChIP-Seq data for the H3K4me2 modification as well as RRBS data for brain, liver, testis, spermatocytes and spermatids. RNA-seq, ChIP-seq and RRBS data were generated from the same individual or pool of individuals, in the case of purified cells. RNA€�Seq data for purified sertoli cells, spermatogonia, spermatocytes, spermatids and spermatozoa
Experiment type
RNA-seq of coding RNA 
Contacts
Magali Soumillon <mag.soumillon@gmail.com>, Anamaria Necsulea, Andreas Gnirke, Bernard de Massy, David Brawand, Henrik Kaessmann, Hongcang Gu, Manuela Weier, Maria Kokkinaki, Martin Dym, Pauline Barthes, Serge Nef, Tarjei S Mikkelsen, Xiaolan Zhang
Citation
Cellular source and mechanisms of high transcriptome complexity in the mammalian testis. Soumillon M, Necsulea A, Weier M, Brawand D, Zhang X, Gu H, Barthes P, Kokkinaki M, Nef S, Gnirke A, Dym M, de Massy B, Mikkelsen TS, Kaessmann H. , Europe PMC 23791531
MINSEQE
Exp. designProtocolsVariablesProcessedSeq. reads
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