Yi Li, Zou Bing, Xie Liping, Zhang Rongqing
Ministry of Education Key Laboratory of Protein Sciences, School of Life Sciences, Tsinghua University, Beijing 100084, P. R. China.
ACS Omega. 2022 Jun 30;7(27):23624-23633. doi: 10.1021/acsomega.2c02230. eCollection 2022 Jul 12.
Matrix proteins play critical roles in regulating the prismatic and nacreous layer formation in the shell. However, due to the dearth of experiments, their specific roles during shell formation are still unclear. In this study, a new method to detect the content of Sr in the nacreous layer (DCSr-NL), which can semiquantitatively measure the nacreous growth rate, has been proposed. experiments show that during crystallization, the Sr element can replace Ca partially, resulting in isomorphism. experiments show that the best labeling conditions are when the Sr/Ca in seawater is 0.3, at 24 °C, and at 4 days of culture. Although a surface morphological difference in the inner layer of nacre is seldom detected by scanning electron microscopy (SEM), knockdown of the classical gene nacrein or unknown gene NU9, combined with DCSr-NL, shows that both significantly decrease the nacreous layer formation rate. The knockdown of the classical gene Pif177 or unknown genes NU3 or MRPN affects the surface morphology and decreases the nacreous layer formation rate. In general, thanks to DCSr-NL, we can efficiently analyze the growth rate of the nacre with or without morphological changes by SEM, and it is of considerable significance for exploring the target gene's function in forming the nacre .
基质蛋白在调节贝壳棱柱层和珍珠层形成过程中发挥着关键作用。然而，由于实验缺乏，它们在贝壳形成过程中的具体作用仍不清楚。在本研究中，提出了一种检测珍珠层中锶含量的新方法（DCSr-NL），该方法可以半定量地测量珍珠层的生长速率。实验表明，在结晶过程中，锶元素可以部分替代钙，从而导致同晶型现象。实验表明，最佳标记条件是海水 Sr/Ca 为 0.3、温度为 24℃且培养 4 天。尽管通过扫描电子显微镜（SEM）很少检测到珍珠层内层的表面形态差异，但经典基因 nacrein 或未知基因 NU9 的敲低，结合 DCSr-NL，表明两者均显著降低珍珠层形成速率。经典基因 Pif177 或未知基因 NU3 或 MRPN 的敲低会影响表面形态并降低珍珠层形成速率。总体而言，借助 DCSr-NL，我们可以通过 SEM 有效分析有无形态变化时珍珠层的生长速率，这对于探索目标基因在珍珠形成中的功能具有相当重要的意义。
