Transcriptomic analysis of dead end knockout testis reveals germ cell and gonadal somatic factors in Atlantic salmon
Transcriptomic analysis of dead end knockout testis reveals germ cell and gonadal somatic factors in Atlantic salmon
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DOI：
10.1186/s12864-020-6513-4
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发表时间：
2020-01-30
期刊：
影响因子：
4.4
通讯作者：
Wargelius, Anna
中科院分区：
文献类型：
--
作者：
Kleppe, Lene;Edvardsen, Rolf Brudvik;Wargelius, Anna
背景可持续挑战目前正在阻碍三文鱼产量的增加。使用不育鲑鱼可以解决性早熟和基因导入问题，从养殖的逃逸到野生种群。最近，不育鲑鱼是通过敲除生殖细胞特异性死胡同(DND)而产生的。可以采用几种方法来抑制DND功能，包括基因敲除、敲除或免疫。由于针对体内已存在的基因产物开发成功的治疗方法具有挑战性，因此正在探索替代靶点。生殖细胞被性腺体细胞包围，并依赖于性腺体细胞。靶向性腺体细胞生存所必需的基因可能是不孕症治疗的良好替代靶点。我们的目的是鉴定和表征大西洋鲑鱼中新的生殖细胞和性腺体细胞因子。结果首次分析了无生殖细胞(GCF)/DND基因敲除和野生型(WT)鲑鱼睾丸的RNA测序数据，寻找在生殖细胞或性腺体细胞中优先表达的基因。为了排除性腺外表达的基因，我们的数据集与可用的多组织转录组数据合并。我们鉴定了389个性腺特异基因，其中194个在生殖细胞中优先表达，11个仅在性腺体细胞中表达。有趣的是，在11个性腺体细胞转录本中，有5个代表了编码分泌转化生长因子-β因子的基因；GSDF、inha、node和两个类似Bmp6的基因，都是代表性的疫苗靶标。其中，GSDF和INHA的转录水平最高。进一步证实GSDF和INHA的表达是性腺特异性的，其空间表达分别局限于卵巢和睾丸的颗粒细胞和支持细胞。最后，我们发现inha在卵巢和睾丸组织中的表达随着青春期的增加而增加，而GSDF在卵巢和睾丸组织中的表达在青春期分别保持不变或下降。结论本研究提供了鲑鱼睾丸组织中生殖细胞和非生殖细胞的转录组数据。我们提供了一个新的和已知的生殖细胞和性腺体细胞特异性转录本的列表，并表明两个高度活跃的性腺体细胞分泌的转化生长因子-β因子，GSDF和INHA，位于颗粒细胞和支持细胞中。
Background Sustainability challenges are currently hampering an increase in salmon production. Using sterile salmon can solve problems with precocious puberty and genetic introgression from farmed escapees to wild populations. Recently sterile salmon was produced by knocking out the germ cell-specific dead end (dnd). Several approaches may be applied to inhibit Dnd function, including gene knockout, knockdown or immunization. Since it is challenging to develop a successful treatment against a gene product already existing in the body, alternative targets are being explored. Germ cells are surrounded by, and dependent on, gonadal somatic cells. Targeting genes essential for the survival of gonadal somatic cells may be good alternative targets for sterility treatments. Our aim was to identify and characterize novel germ cell and gonadal somatic factors in Atlantic salmon. Results We have for the first time analysed RNA-sequencing data from germ cell-free (GCF)/dnd knockout and wild type (WT) salmon testis and searched for genes preferentially expressed in either germ cells or gonadal somatic cells. To exclude genes with extra-gonadal expression, our dataset was merged with available multi-tissue transcriptome data. We identified 389 gonad specific genes, of which 194 were preferentially expressed within germ cells, and 11 were confined to gonadal somatic cells. Interestingly, 5 of the 11 gonadal somatic transcripts represented genes encoding secreted TGF-beta factors; gsdf, inha, nodal and two bmp6-like genes, all representative vaccine targets. Of these, gsdf and inha had the highest transcript levels. Expression of gsdf and inha was further confirmed to be gonad specific, and their spatial expression was restricted to granulosa and Sertoli cells of the ovary and testis, respectively. Finally, we show that inha expression increases with puberty in both ovary and testis tissue, while gsdf expression does not change or decreases during puberty in ovary and testis tissue, respectively. Conclusions This study contributes with transcriptome data on salmon testis tissue with and without germ cells. We provide a list of novel and known germ cell- and gonad somatic specific transcripts, and show that the expression of two highly active gonadal somatic secreted TGF-beta factors, gsdf and inha, are located within granulosa and Sertoli cells.