Comparative Analysis of Testis Transcriptomes from Triploid and Fertile Diploid Cyprinid Fish
Comparative Analysis of Testis Transcriptomes from Triploid and Fertile Diploid Cyprinid Fish
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三倍体与可育二倍体鲤鱼睾丸转录组的比较分析
DOI：
10.1095/biolreprod.114.125609
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发表时间：
2015-04-01
影响因子：
3.6
通讯作者：
Liu, Shaojun
中科院分区：
文献类型：
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作者：
Xu, Kang;Wen, Ming;Liu, Shaojun
鱼类的繁殖力是鱼类育种的关键因素。RNA-seq广泛用于高通量测序，并提供了一种快速方法来检查生物过程的分子机制。为了探索生育力相关的分子机制，我们获得了二倍体和三倍体鲤科鱼类的睾丸转录组，并测试了睾丸中的差异表达基因（DEG）。在三倍体和二倍体鱼中共检测到6730个差异表达的转录本。在这些转录本中，与二倍体鱼相比，2428个转录本在三倍体鱼中表现出降低的表达，4302个转录本在三倍体鱼中过表达。功能分析表明，部分基因相关的生殖，发育和运动过程，以及轴丝，在三倍体鱼相对于二倍体鱼的差异表达。途径分析表明，泛醌和其他萜类醌生物合成途径和凋亡途径的基因表达水平的变化在三倍体雄鱼不育中起着核心作用。一系列与精子鞭毛组装和运动相关的基因（DNAH、DNAL 1、IFTs和DNAAF 1）以及睾丸特异性候选标记（Tcte 1、Tekt 1、Tekt 4、Spag 17、Spag 5、Spag 9a、Spag 1b和Spef 2）在三倍体鱼睾丸中表达水平较低。我们验证了这些DEG在三倍体鱼使用定量PCR定量表达的8个代表性的基因。此外，276个假定的转录因子，6染色质重塑因子，和35个转录辅因子在三倍体鱼相比，二倍体表现出差异表达。本研究提供了深入了解导致雄性三倍体鱼不育的调控机制。
The fertility of fish is a key factor in fish breeding. RNA-seq is widely used in high-throughput sequencing and provides a rapid method to examine the molecular mechanisms underlying a biological process. To probe fertility-related molecular mechanisms, we obtained testis transcriptomes from diploid and triploid cyprinid fish and tested for differentially expressed genes (DEGs) in the testis. A total of 6730 transcripts were differentially expressed between the triploid and diploid fish. In these transcripts, 2428 transcripts showed reduced expression and 4302 transcripts were overexpressed in triploid fish compared to the diploid fish. Functional analyses revealed that partial genes related to reproductive, developmental, and locomotion processes, and the axoneme, were differentially expressed in triploid fish relative to diploid fish. Pathway analysis indicated that variations in the gene expression levels of the "ubiquinone and other terpenoid-quinone biosynthesis pathway'' and the "apoptotic pathway'' played a central role in the sterility of triploid male fish. A series of genes (DNAHs, DNAL1, IFTs, and DNAAF1) associated with sperm flagellar assembly and motility, and testis-specific candidate markers (Tcte1, Tekt1, Tekt4, Spag17, Spag5, Spag9a, Spag1b, and Spef2), had low expression levels in the testis of triploid fish. We validated these DEGs in triploid fish using quantitative PCR to quantify expression of eight representative genes. Furthermore, 276 putative transcription factors, 6 chromatin remodeling factors, and 35 transcription cofactors exhibited differential expression in triploid compared to diploid fish. This study provides insight into the regulatory mechanisms causing sterility in male triploid fish.