三角帆蚌不同蚌龄外套膜细胞的增殖能力及其矿化功能

研究了淡水珍珠贝——三角帆蚌(Hyriopsis cumingii)在不同蚌龄下外套膜组织的细胞增殖及生物矿化相关因子活性。选择5组不同蚌龄三角帆蚌(0.5龄、1龄、2龄、3龄、4龄),通过流式细胞技术分析了外套膜细胞的增殖指数(proliferation index,PrI)和胞内Ca~(2+)浓度,并应用实时荧光定量PCR检测了与生物矿化相关的碳酸酐酶(carbonic anhydrase, CA)、碱性磷酸酶(alkaline phosphatase, ALP)基因的表达并测定其酶活性。结果表明, 2龄蚌的外套膜细胞PrI及胞内Ca~(2+)浓度显著高于其他蚌龄(P0.05);生物矿化相关基因在不同蚌龄的外套膜组织中表达量不同, 1龄三角帆蚌CA基因表达量和CA酶活性最高(P0.05), ALP基因表达量和ALP酶活在0.5龄三角帆蚌中显著高于其他蚌龄(P0.05)。本研究旨为深入探讨三角帆蚌外套膜细胞增殖能力及人工育珠过程中供体蚌的选择奠定基础。

Proliferative capacity and biomineralization of mantle cells in Hyriopsis cumingii of different ages

The mantle is an indispensable tissue for pearl cultivation, but the biological activity (cell proliferation and biomineralization) of donor mussel mantle of different ages has not been systematically explored. In this study, was used to investigate the proliferative activity and biomineralization capacity in mantle cells in different years. In this experiment, five groups of were selected by age, at six months and one, two, three, and four years. Flow cytometry was used to detect the cell cycle of mantle cells of different ages and to calculate the cell proliferation index. Fluo-3/AM, a calcium fluorescence probe, combines with intracellular Ca²⁺ to produce fluorescence. The average fluorescence intensity of mantle cells of different ages was measured by flow cytometry fluorescein isothiocyanate (FITC) channel, which represented intracellular calcium concentration. Quantitative real-time PCR (qRT-PCR) was used to analyze expression of biomineralization related genes. Biomineralization-related genes included carbonic anhydrase gene (). The enzyme activities of ALP and CA were determined chemically and by ELISA, respectively. The aim of this study was to determine the suitable age of with fast proliferation and strong biomineralization ability of mantle cells. The results showed that:(a) Under the same environment, the proliferation index of two-year-old mantle cells was the largest (i.e., fastest proliferation). (b) The average fluorescence intensity of the two-year-old was the highest, in other words, the concentration of calcium ion in the mantle cells of the two-year-old gene in six-month-old mussels was significantly higher than that in older mussels (<0.05), followed by the one-year-old mussels and the two-year-old mussels. There was no significant difference between the three and four-year-old mussels, with the lowest gene expression level (<0.05). (d) The expression level of was different in mantle tissues in mussels of different ages. The highest expression level (<0.05) was found in one-year-old mussels, followed by the two-year-old mussels, and the lowest expression level (>0.05) was found in six-month-old, three and four-year-old mussels. The activity of CA was different in mantle tissue of different mussels and the activity of CA was the highest in one-year-old mussels (<0.05). Finally, it was concluded that the proliferation of mantle cells of the two-year-old mussels was the strongest and the intracellular calcium concentration was the highest. This study provides a foundation for further research on the proliferation of mantle cells in and the selection of donor mussel age for pearl culture.