Key Laboratory of Exploration and Utilization of Aquatic Genetic Resources, Ministry of Education, Shanghai Ocean University, Shanghai, 201306, China.
Jiangsu Key Laboratory of Marine Biotechnology, Jiangsu Ocean University, Lianyungang, 222005, China.
Mar Biotechnol (NY). 2020 Oct;22(5):696-705. doi: 10.1007/s10126-020-09993-0. Epub 2020 Sep 4.
The razor clam, Sinonovacula constricta, is an important economic marine shellfish, and its larval development involves obvious morphological and physiological changes. MicroRNA plays a key role in the physiological changes of the organism through regulating targeted mRNA. This study performed miRNA-mRNA sequencing for eight different developmental stages of S. constricta using Illumina sequencing. A total of 2156 miRNAs were obtained, including 2069 known miRNAs and 87 novel miRNAs. In addition, target genes were predicted for key miRNAs differentially expressed between adjacent development samples by integrating the mRNA transcriptome. Further analysis revealed that the differentially expressed genes were enriched in complement activation, alternative pathways, translation, and negative regulation of monocyte molecular protein-1 production. KEGG pathway annotation showed significant enrichment in the regulation of the ribosome, phagosome, tuberculosis and fluid shear stress, and atherosclerosis. Ten mRNAs and ten miRNAs that are related to larval metamorphosis were identified using real-time PCR. Furthermore, the double luciferase experiment validated the negative regulatory relationship between miR-133 and peroxisome proliferator-activated receptor-γ (PPAR-γ). These results indicated that the target genes regulated by these differentially expressed miRNAs may play an important regulatory role in the metamorphosis development of S. constricta.
菲律宾蛤仔是一种重要的经济贝类，其幼虫发育过程涉及明显的形态和生理变化。miRNA 通过调控靶向 mRNA 在生物体的生理变化中发挥关键作用。本研究利用 Illumina 测序对菲律宾蛤仔的 8 个不同发育阶段进行了 miRNA-mRNA 测序。共获得 2156 个 miRNA，包括 2069 个已知 miRNA 和 87 个新 miRNA。此外，通过整合 mRNA 转录组，对相邻发育样本中差异表达的关键 miRNA 的靶基因进行了预测。进一步分析表明，差异表达基因富集在补体激活、替代途径、翻译和单核细胞分子蛋白 1 产生的负调控。KEGG 通路注释显示，核糖体、吞噬体、结核和流体切应力以及动脉粥样硬化的调控显著富集。通过实时 PCR 鉴定出 10 个与幼虫变态相关的 mRNAs 和 10 个 miRNAs。此外，双荧光素酶实验验证了 miR-133 与过氧化物酶体增殖物激活受体-γ（PPAR-γ）之间的负调控关系。这些结果表明，这些差异表达 miRNA 调控的靶基因可能在菲律宾蛤仔变态发育中发挥重要的调控作用。
