Unknown,Transcriptomics,Genomics,Proteomics

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The Multicopy Gene Sly Represses the Sex Chromosomes in the Male Mouse Germline after Meiosis

ABSTRACT: Studies of mice with Y chromosome long arm deficiencies suggest that the male-specific region (MSYq) encodes information required for sperm differentiation and postmeiotic sex chromatin repression (PSCR). Several genes have been identified on MSYq, but because they are present in more than 40 copies each, their functions cannot be investigated using traditional gene targeting. Here, we generate transgenic mice producing small interfering RNAs that specifically target the transcripts of the MSYq-encoded multicopy gene Sly (Sycp3-like Y-linked). Microarray analyses performed on these Sly-deficient males and on MSYq-deficient males show a remarkable up-regulation of sex chromosome genes in spermatids. Two sets of samples were examined: (1) Total RNA isolated from separated round spermatids of young adult males (8 weeks postpartum) with Yq deletions or shSLY targeted knockdown, compared to age- and strain-matched controls. (2) Total RNA isolated from whole testis of juvenile males (17 days post partum) with shSLY targeted knockdown, compared to age- and strain-matched controls.

ORGANISM(S): Mus musculus

SUBMITTER: Peter Ellis

PROVIDER: E-GEOD-29198 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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The multicopy gene Sly represses the sex chromosomes in the male mouse germline after meiosis.

Cocquet Julie J Ellis Peter J I PJ Yamauchi Yasuhiro Y Mahadevaiah Shantha K SK Affara Nabeel A NA Ward Monika A MA Burgoyne Paul S PS

PLoS biology 20091117 11

Studies of mice with Y chromosome long arm deficiencies suggest that the male-specific region (MSYq) encodes information required for sperm differentiation and postmeiotic sex chromatin repression (PSCR). Several genes have been identified on MSYq, but because they are present in more than 40 copies each, their functions cannot be investigated using traditional gene targeting. Here, we generate transgenic mice producing small interfering RNAs that specifically target the transcripts of the MSYq- ...[more]

PMID: 19918361

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Project description:Understanding the extent of genomic transcription and its functional relevance is a central goal in genomics research. However, detailed genomewide 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 RNAseq 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 postmeiotic 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 genomewide 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 RNASeq 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. RNASeq data for purified sertoli cells, spermatogonia, spermatocytes, spermatids and spermatozoa

Dataset Information

The Multicopy Gene Sly Represses the Sex Chromosomes in the Male Mouse Germline after Meiosis

Publications

The multicopy gene Sly represses the sex chromosomes in the male mouse germline after meiosis.

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