背景:在生命早期阶段,涉及头足类发育的基因及其与孵化和存活的关联已得到广泛研究。然而,很少有研究调查了东亚普通章鱼(章鱼sinensis)的副转录组。
目的:本研究旨在通过RNA测序(RNA-seq)鉴定与中华绒螯蟹胚胎发育和孵化相关的基因,并验证与不同胚胎阶段最相关的基因。
方法:使用孵化后和孵化后25天(dph)的RNA样品构建cDNA文库。将来自单个样品的干净读数与参考O.sinensis数据库进行比对,以鉴定0-和25-dph副细菌文库之间的差异表达基因(DEGs)。实时定量聚合酶链反应(RT-qPCR)用于补充胚胎发生发育阶段的RNA-seq数据。
结果:总共注释了12,597个转录本,并在0-和25-dphO之间鉴定了5,468个DEG。在25-dph副翼中,包括2,715个上调和2,753个下调的转录本。几个关键的DEGs与跨膜运输有关,脂质生物合成,单加氧酶活性,脂质运输,神经肽信号,转录调节,在孵化后发育过程中,蛋白质-半胱氨酸S-棕榈酰转移酶活性。RT-qPCR分析进一步显示SLC5A3A,受精后20和/或30天(dpf)胚胎中的ABCC12和NPC1转录物明显高于10-dpf胚胎(p<0.05)。
结论:转录组谱为了解胚胎发育提供了分子靶标,孵化,和中华副甲的存活,并提高章鱼的生产。
BACKGROUND: The genes involved in cephalopod development and their association with hatching and survival during early life stages have been extensively studied. However, few studies have investigated the paralarvae transcriptome of the East Asian common octopus (Octopus sinen sis).
OBJECTIVE: This study aimed to identify the genes related to embryonic development and hatching in O. sinensis using RNA sequencing (RNA-seq) and verify the genes most relevant to different embryonic stages.
METHODS: RNA samples from hatched and 25 days post-hatching (dph) O. sinensis paralarvae were used to construct cDNA libraries. Clean reads from individual samples were aligned to the reference O. sinensis database to identify the differentially expressed genes (DEGs) between the 0- and 25-dph paralarvae libraries. Real-time quantitative polymerase chain reaction (RT-qPCR) was used to supplement the RNA-seq data for embryogenic developmental stages.
RESULTS: A total of 12,597 transcripts were annotated and 5,468 DEGs were identified between the 0- and 25-dph O. sinensis paralarvae, including 2,715 upregulated and 2,753 downregulated transcripts in the 25-dph paralarvae. Several key DEGs were related to transmembrane transport, lipid biosynthesis, monooxygenase activity, lipid transport, neuropeptide signaling, transcription regulation, and protein-cysteine S-palmitoyltransferase activity during the post-hatching development of O. sinensis paralarvae. RT-qPCR analysis further revealed that SLC5A3A, ABCC12, and NPC1 transcripts in 20 and/or 30 days post-fertilization (dpf) embryos were significantly higher (p < 0.05) than those in 10-dpf embryos.
CONCLUSIONS: Transcriptome profiles provide molecular targets to understand the embryonic development, hatching, and survival of O. sinensis paralarvae, and enhance octopus production.