Expression dynamics, relationships, and transcriptional regulations of diverse transcripts in mouse spermatogenic cells
Expression dynamics, relationships, and transcriptional regulations of diverse transcripts in mouse spermatogenic cells
复制标题
小鼠生精细胞中不同转录物的表达动态、关系和转录调控。
DOI：
10.1080/15476286.2016.1218588
复制
发表时间：
2016-01-01
期刊：
影响因子：
4.1
通讯作者：
Han, Chunsheng
中科院分区：
文献类型：
--
作者：
Lin, Xiwen;Han, Miao;Han, Chunsheng
在后生动物的所有组织中，精巢的转座体表现出最高的多样性和特异性。然而，其组成和动态在精子发生过程中尚未完全了解。在此，我们在小鼠生精细胞中鉴定了20，639种信使RNA（messagerRNAs，mRNAs）、7，168种长链非编码RNA（longnoncodingRNAs，lncRNAs）和15，101种环状RNA（circRNAs），并发现其中许多在睾丸中特异表达。lncRNA比mRNA具有更高的睾丸特异性。在所有阶段，mRNA通常比lncRNA更丰富，线性转录物比circRNA更丰富。我们发现circRNA和piRNA的产生是高度调节的，而不是随机过程。基于一个小规模的功能筛选实验的结果，使用培养的小鼠精原干细胞，许多进化上保守的lncRNA可能在精子发生中发挥作用。睾丸特异性m/lncRNA基因的启动子中富含典型类型的转录因子结合位点。CREM和RFX 2是精子发生的2个关键TF，通过使用ChIP芯片分析和RFX 2敲除生精细胞的RNA-seq进一步验证了CREM和RFX 2的靶基因。我们的研究结果有助于目前的生精细胞的转录组的复杂性的理解，并提供了一个有价值的资源，许多候选基因可能会被选择为进一步的功能研究。
Among all tissues of the metazoa, the transcritpome of testis displays the highest diversity and specificity. However, its composition and dynamics during spermatogenesis have not been fully understood. Here, we have identified 20,639 message RNAs (mRNAs), 7,168 long non-coding RNAs (lncRNAs) and 15,101 circular RNAs (circRNAs) in mouse spermatogenic cells, and found many of them were specifically expressed in testes. lncRNAs are significantly more testis-specific than mRNAs. At all stages, mRNAs are generally more abundant than lncRNAs, and linear transcripts are more abundant than circRNAs. We showed that the productions of circRNAs and piRNAs were highly regulated instead of random processes. Based on the results of a small-scale functional screening experiment using cultured mouse spermatogonial stem cells, many evolutionarily conserved lncRNAs are likely to play roles in spermatogenesis. Typical classes of transcription factor binding sites are enriched in the promoters of testis-specific m/lncRNA genes. Target genes of CREM and RFX2, 2 key TFs for spermatogenesis, were further validated by using ChIP-chip assays and RNA-seq on RFX2-knockout spermatogenic cells. Our results contribute to the current understanding of the transcriptomic complexity of spermatogenic cells and provide a valuable resource from which many candidate genes may be selected for further functional studies.