Musacchia Francesco, Vasilev Filip, Borra Marco, Biffali Elio, Sanges Remo, Santella Luigia, Chun Jong Tai
Department of Biology and Evolution of Marine Organisms, Stazione Zoologica Anton Dohrn, Napoli, Italy.
The Molecular Biology and Bioinformatics Unit, Stazione Zoologica Anton Dohrn, Napoli, Italy.
PLoS One. 2017 Sep 5;12(9):e0184090. doi: 10.1371/journal.pone.0184090. eCollection 2017.
Starfish have been instrumental in many fields of biological and ecological research. Oocytes of Astropecten aranciacus, a common species native to the Mediterranean Sea and the East Atlantic, have long been used as an experimental model to study meiotic maturation, fertilization, intracellular Ca2+ signaling, and cell cycle controls. However, investigation of the underlying molecular mechanisms has often been hampered by the overall lack of DNA or protein sequences for the species. In this study, we have assembled a transcriptome for this species from the oocytes, eggs, zygotes, and early embryos, which are known to have the highest RNA sequence complexity. Annotation of the transcriptome identified over 32,000 transcripts including the ones that encode 13 distinct cyclins and as many cyclin-dependent kinases (CDK), as well as the expected components of intracellular Ca2+ signaling toolkit. Although the mRNAs of cyclin and CDK families did not undergo significant abundance changes through the stages from oocyte to early embryo, as judged by real-time PCR, the transcript encoding Mos, a negative regulator of mitotic cell cycle, was drastically reduced during the period of rapid cleavages. Molecular phylogenetic analysis using the homologous amino acid sequences of cytochrome oxidase subunit I from A. aranciacus and 30 other starfish species indicated that Paxillosida, to which A. aranciacus belongs, is not likely to be the most basal order in Asteroidea. Taken together, the first transcriptome we assembled in this species is expected to enable us to perform comparative studies and to design gene-specific molecular tools with which to tackle long-standing biological questions.
海星在生物和生态研究的许多领域都发挥了重要作用。橙黄阳遂足(Astropecten aranciacus)是一种原产于地中海和东大西洋的常见海星,其卵母细胞长期以来一直被用作研究减数分裂成熟、受精、细胞内Ca2+信号传导和细胞周期控制的实验模型。然而,由于该物种总体上缺乏DNA或蛋白质序列,对其潜在分子机制的研究常常受到阻碍。在本研究中,我们从卵母细胞、卵子、受精卵和早期胚胎中组装了该物种的转录组,已知这些细胞具有最高的RNA序列复杂性。转录组注释识别出超过32000个转录本,包括编码13种不同细胞周期蛋白和同样数量的细胞周期蛋白依赖性激酶(CDK)的转录本,以及细胞内Ca2+信号传导工具包的预期成分。尽管通过实时PCR判断,细胞周期蛋白和CDK家族的mRNA在从卵母细胞到早期胚胎的各个阶段没有发生显著的丰度变化,但编码有丝分裂细胞周期负调节因子Mos的转录本在快速分裂期间急剧减少。使用橙黄阳遂足和其他30种海星物种的细胞色素氧化酶亚基I的同源氨基酸序列进行的分子系统发育分析表明,橙黄阳遂足所属的柱体海星目不太可能是海星纲中最基部的目。综上所述,我们在该物种中组装的首个转录组有望使我们能够进行比较研究,并设计基因特异性分子工具来解决长期存在的生物学问题。