三角帆蚌不同蚌龄外套膜细胞的增殖能力及其矿化功能
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作者单位:

1. 上海海洋大学, 农业部淡水水产种质资源重点实验室, 上海 201306;
2. 上海海洋大学, 水产种质资源发掘与利用教育部重点实验室, 上海 201306;
3. 上海海洋大学, 水产科学国家级实验教学示范中心, 上海 201306

作者简介:

曹玉香(1993-),女,硕士研究生,从事水生动物细胞分子生物技术研究.E-mail:yxcao77@126.com

通讯作者:

中图分类号:

S917

基金项目:

国家自然科学基金项目(31201991);国家重点研发计划项目(2018YFD0901406).


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

1. Key Laboratory of Freshwater Aquatic Genetic Resources, Ministry of Agriculture;Shanghai Ocean University, Shanghai 201306, China;
2. Key Laboratory of Exploration and Utilization of Aquatic Genetic Resources, Ministry of Education;Shanghai Ocean University, Shanghai 201306, China;
3. National Demonstration Centre for Experimental Fisheries Science Education, Shanghai Ocean University, Shanghai 201306, China

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    摘要:

    研究了淡水珍珠贝——三角帆蚌()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Ca2+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Ca2+浓度显著高于其他蚌龄(<0.05),<0.05)。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。

    Abstract:

    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.

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曹玉香,陆阿利,李文娟,施志仪,陶家康.三角帆蚌不同蚌龄外套膜细胞的增殖能力及其矿化功能[J].中国水产科学,2020,27(5):516-523
CAO Yuxiang, LU Ali, LI Wenjuan, SHI Zhiyi, TAO Jiakang. Proliferative capacity and biomineralization of mantle cells in Hyriopsis cumingii of different ages[J]. Journal of Fishery Sciences of China,2020,27(5):516-523

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  • 在线发布日期: 2020-05-15
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