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RNA-seq reveals an abnormal transcriptome as a result of DDX20 loss in spermatogonia.

ABSTRACT: RNA-seq reveals an abnormal transcriptome as a result of DDX20 loss in spermatogonia.

PROVIDER: PRJNA900146 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR22252975_1.fastq.gz	Fastqsanger.gz
	SRR22252975_2.fastq.gz	Fastqsanger.gz
	SRR22252976_1.fastq.gz	Fastqsanger.gz
	SRR22252976_2.fastq.gz	Fastqsanger.gz
	SRR22252977_1.fastq.gz	Fastqsanger.gz

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RNA-seq reveals an abnormal transcriptome as a result of DDX20 loss in spermatogonia.

Project description:To investigate the role of DDX20 in spermatogenesis, we generated Ddx20 flox/flox Ddx4-Cre mice to make a germ cell-specific Ddx20 knockout, and isolated spermatogonia from four-day-old mouse testes by THY1 magnetic beads.We then performed gene expression profiling analysis using data obtained from RNA-seq of THY1 + spermatogonia.

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SRSF10 is essential for progenitor spermatogonia expansion by regulating alternative splicing

Project description:Alternative splicing expands the proteome complexity and plays essential roles in tissue development and human diseases. However, how spermatogenesis is regulated by alternative splicing remains largely unknown. Here, using germ cell-specific knockout mouse model, we demonstrate that the splicing factor Srsf10 is essential for spermatogenesis and male fertility. Depletion of Srsf10 in germ cells had little effect on the formation of SSCs but impeded the expansion of progenitor spermatogonia, leading to the failure of spermatogonia differentiation and meiosis initiation. This was evidenced by the decreased expression of progenitor cell markers in bulk RNA-seq, and much less progenitor and differentiating spermatogonia in single-cell RNA-seq data. Furthermore, the expression of genes involved in cell cycle was abnormal in all subtypes of spermatogonia identified in single-cell RNA-seq data. Notably, using isolated spermatogonia, we found that Srsf10 depletion disturbed the alternative splicing of hundreds of genes, which were preferentially associated with cell cycle, mitotic cell cycle checkpoint and germ cell development, including Dazl, Kit, Ret, Sycp1, Nasp and Bora. These data suggest that SRSF10 is critical for the expansion of progenitor spermatogonia by regulating alternative splicing, expanding our understanding of the novel mechanism underlying spermatogenesis.

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Single-cell Gene Expression Analysis Reveals Transcriptional Diversity among Human Spermatogonia

Project description:Two individual human OCT4-positive spermatogonia were selected for global transcriptional capture using shallow RNA-seq. Successful cDNA synthesis and amplification was confirmed as reads mapped primarily to exons. It has to be noted though, that a comparision of cell to cell data is not possible as coverage of the whole transcriptome was too low, approximately <1.0x coverage. Nonetheless, RNA-seq data provided first information of highly abundant transcripts in individual human spermatogonia. Spermatogonia-specific expression of selected candidate genes with high RPKM values including DDX5, EEF1A1 and TSPY1 was confirmed by immunohistochemistry.

2017-01-15 | GSE91063 | GEO

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