Fishery College, Guangdong Ocean University, Zhanjiang, 524025, China.
Pearl Breeding and Processing Engineering Technology Research Centre of Guangdong Province, Zhanjiang, 524088, China.
Mar Biotechnol (NY). 2023 Jun;25(3):428-437. doi: 10.1007/s10126-023-10216-5. Epub 2023 May 29.
MicroRNAs (miRNAs) are small noncoding RNAs that regulate gene expression via the recognition of their target messenger RNAs. MiR-10a-3p plays an important role in the process of ossification. In this study, we obtained the precursor sequence of miR-10a-3p in the pearl oyster Pinctada fucata martensii (Pm-miR-10a-3p) and verified its sequence by miR-RACE technology, and detected its expression level in the mantle tissues of the pearl oyster P. f. martensii. Pm-nAChRsα and Pm-NPY were identified as the potential target genes of Pm-miR-10a-3p. After the over-expression of Pm-miR-10a-3p, the target genes Pm-nAChRsα and Pm-NPY were downregulated, and the nacre microstructure became disordered. The Pm-miR-10a-3p mimic obviously inhibited the luciferase activity of the 3' untranslated region of the Pm-NPY gene. When the interaction site was mutated, the inhibitory effect disappeared. Our results suggested that Pm-miR-10a-3p participates in nacre formation in P. f. martensii by targeting Pm-NPY. This study can expand our understanding of the mechanism of biomineralization in pearl oysters.
微小 RNA（miRNAs）是通过识别其靶信使 RNA 来调节基因表达的小非编码 RNA。miR-10a-3p 在骨化过程中发挥着重要作用。本研究在马氏珠母贝（Pinctada fucata martensii）中获得了 miR-10a-3p 的前体序列（Pm-miR-10a-3p），并通过 miR-RACE 技术验证了其序列，检测了其在马氏珠母贝(Pinctada fucata martensii) 套膜组织中的表达水平。鉴定了 Pm-nAChRsα 和 Pm-NPY 为 Pm-miR-10a-3p 的潜在靶基因。过表达 Pm-miR-10a-3p 后，靶基因 Pm-nAChRsα 和 Pm-NPY 下调，珍珠贝的珍珠层微观结构变得紊乱。Pm-miR-10a-3p 模拟物明显抑制了 Pm-NPY 基因 3'非翻译区的荧光素酶活性。当相互作用位点发生突变时，抑制作用消失。我们的结果表明，Pm-miR-10a-3p 通过靶向 Pm-NPY 参与马氏珠母贝的珍珠层形成。本研究可以扩展我们对珍珠贝生物矿化机制的理解。
