A novel meiosis-related lncRNA, Rbakdn, contributes to spermatogenesis by stabilizing Ptbp2
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ABSTRACT:
PROVIDER: PRJNA761683 | ENA |
REPOSITORIES: ENA
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Project description:Spermatocyte meiosis is the cornerstone of mammalian production. Thousands of long noncoding RNAs (lncRNAs) have been reported to be functional in various cellular processes, but the function of lncRNAs in meiosis remains largely unknown. Here, we profiled lncRNAs in spermatocytes at stage I of meiosis and identified a testis-specific lncRNA, Rbakdn, as a vital regulator of meiosis. Rbakdn is dynamically expressed during meiosis I, and Rbakdn knockdown inhibits meiosis in vitro. Furthermore, Rbakdn knockdown in testes in mice by intratesticular injection disturbs meiosis, reduces testicular volume, and increases apoptosis of spermatocytes, resulting in vacuolation of the seminiferous tubules. Rbakdn can bind to Ptbp2, an RNA-binding protein that is important in the regulation of the alternative splicing of many genes in spermatogenesis. Rbakdn knockdown leads to a decrease in Ptbp2 through the ubiquitination degradation pathway, indicating that Rbakdn maintains the stability of Ptbp2. In conclusion, our study identified an lncRNA, Rbakdn, with a crucial role in meiosis.
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Project description:Male germ cell meiosis is essential for generating haploid spermatozoa in mice. Here, we investigate the essential role of DIS3 in male germ cell meiosis in mice. Conditional inactivation of DIS3 in spermatocytes with Stra8-cre transgenic mice have severely impaired meiotic progression, which results in defective meiosis and spermatogenesis. RNA-seq analysis reveals that Dis3 deficiency causes significant dysregulation of the expression of transcripts in mutant testes. Meiosis-associated genes are significantly decreased in the absence of DIS3. Therefore, we show that DIS3 ribonuclease plays a critical role in germ cell meiosis during spermatogenesis in mice.
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Project description:Alternative splicing (AS) plays key roles in the specialization of cell functions in different tissues and stages of development. In spermatogenic cells, the complexity of AS isoforms exceeds that of most whole tissues, but the regulation, dynamics, and functions of AS in spermatogenesis have remained poorly defined. We previously demonstrated that the RNA binding protein Ptbp2 is essential for cell survival during spermatogenesis; however, the underlying mechanisms were not explored. Here, we demonstrate that Ptbp2 is a critical AS regulator in spermatogenic cells, and controls a functionally-related network of genes involved in germ cell adhesion and protein trafficking to and from the plasma membrane. In parallel, we identify distinct AS programs in different stages of spermatogenesis, and demonstrate an important role for Ptbp2 in stage-specific AS regulation. Collectively, the data provide new insights into the temporal dynamics and importance of AS in mammalian germ cell development, and demonstrate a central role for Ptbp2 in its regulation.
2017-06-20 | GSE79104 | GEO
Project description:Alternative splicing (AS) plays key roles in the specialization of cell functions in different tissues and stages of development. In spermatogenic cells, the complexity of AS isoforms exceeds that of most whole tissues, but the regulation, dynamics, and functions of AS in spermatogenesis have remained poorly defined. We previously demonstrated that the RNA binding protein Ptbp2 is essential for cell survival during spermatogenesis, however the underlying mechanisms were not explored. To investigate this, we generated paired-end RNA-Seq data from wild type (WT) and Ptbp2 conditional knockout (cKO) testes at postnatal day 25 (p25) using Illumina ScriptSeq™ v2 RNA-Seq library preparation kits and sequenced at the CWRU Sequencing Core. Our resulting analyses demonstrate that Ptbp2 is a critical AS regulator in spermatogenic cells, where it controls a functionally-related network of genes involved in germ cell adhesion and protein trafficking to and from the plasma membrane. In parallel, we identified distinct AS programs in different stages of spermatogenesis, and defined a role for Ptbp2 in stage-specific AS regulation. Collectively, the data provide new insights into the importance of AS in mammalian germ cell development and demonstrate a central role for Ptbp2 in its regulation.
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