维生素D_3对三角帆蚌外套膜细胞内Ca^(2+)浓度的影响  

Effect of VD_3 on Intracellular Calcium Ion Concentration in Mantle Cells of Hyriopsis cumingii:An in Vitro Study

作　　者：郝莹莹[1] 施志仪[1] 李文娟[1] 靳雨丽[1] 

机构地区：[1]上海海洋大学农业部水产种质资源与养殖生态重点开放实验室,上海201306

出　　处：《动物学杂志》2011年第3期98-103,共6页Chinese Journal of Zoology

基　　金：上海市科委基础重大项目(No.06aj14003);上海市重点学科水生生物学建设项目(No.S30701);上海市博士后科研资助计划面上项目(No.09R21413200)

摘　　要：采用胰酶消化法获得三角帆蚌(Hyriopsis cumingii)外套膜细胞,用Fluo-3/AM荧光标记技术和激光共聚焦扫描显微镜(confocal laser scanning microscope,CLSM)检测外套膜内外表皮细胞在静息状态下细胞的游离Ca2+浓度及不同浓度维生素D3(VD3)孵育后的外套膜细胞的Ca2+的动态情况。结果表明,未添加VD3(1.25 mmol/L Ca2+)时,外套膜外表皮细胞的Ca2+荧光强度显著高于内表皮细胞内的Ca2+(P<0.05)。在添加不同浓度VD3后,内表皮细胞与外表皮细胞内Ca2+浓度变化趋势一致,表现为细胞内Ca2+沉积量的变化是随着VD3浓度的增大而增大,其中,对照组与添加50 IU/L组差异不显著(P>0.05);当添加浓度为100 IU/L时,与对照组和50 IU/L组差异显著(P<0.05);当添加组浓度达到500IU/L时,与前3组有显著差异(P<0.05);当添加的浓度增大到1 000 IU/L后与500 IU/L组差异不显著(P>0.05),但与其他组均存在显著差异(P<0.05)。本实验为外套膜细胞的钙代谢机制研究提供了实验依据。The mantle cells from Hyriopsis cumingii were obtained by using trypsin digestion.The Ca2＋ concentrations in inner mantle cells and outer mantle cells under the resting condition and the dynamics of Ca2＋concentration in the mantle cells incubated with various concentrations of VD3 were measured by Fluo-3/AM fluorescent labeling and CLSM.The results showed that the average Ca2＋ fluorescence intensity of outer mantle cells was significantly higher than that of inner mantle cells（P0.05）.After adding different concentrations of VD3,the average Ca2＋ fluorescence intensity was enhanced with the increased concentration of VD3 in culture medium,and inner mantle cells and outer mantle cells showed the same change.The difference between the control and 50 IU/L VD3 groups,or between 500 IU/L and 1 000 IU/L groups was not significant（P0.05）,but the difference between the control and another three groups（100 IU/L,500 IU/L and 1 000 IU/L）,or between the 50 IU/L group and another three groups（100 IU/L,500 IU/L and 1 000 IU/L） was significant（P0.05）.The present research may provide theoretical basis for promoting the pearl industry.

关 键 词：三角帆蚌 外套膜细胞Ca2+浓度 维生素D3(VD3) 激光共聚焦扫描显微镜 

分 类 号：S917.4[农业科学—水产科学]