应用激光共聚焦显微技术研究Ca^2＋在三角帆蚌组织内的积累与分布  

Study of Ca^(2+) deposition and distribution in cells from different tissues with laser scanning confocal microscopy in Hyriopsis cumingii

作　　者：李文娟[1] 施志仪[1,2] 郝莹莹[1] 叶显峰 

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 [2]上海市高校水产养殖学E-研究院,上海海洋大学水产与生命学院,上海201306 [3]安徽省芜湖市繁昌县农委,安徽繁昌241200

出　　处：《水产学报》2011年第2期214-220,共7页Journal of Fisheries of China

基　　金：上海市博士后科研资助计划面上项目(09R21413200);上海市科委基础重大项目(06aj14003);上海市重点学科水生生物学建设项目资助(S30701)

摘　　要：为了探讨淡水贝类Ca2+吸收和转运信号传递机理,采用激光共聚焦技术研究了三角帆蚌不同组织细胞在静息状态下细胞内游离Ca2+浓度,以及水体Ca2+浓度对外套膜组织细胞内钙沉积的影响。试验选取10只健康的三角帆蚌,分别获得外套膜外膜、内膜、斧足、腮及内脏团上表皮组织细胞,短期培养后用Fluo-3/AM荧光探针孵育细胞1 h,观察细胞内Ca2+荧光强度。研究结果表明,蚌不同组织细胞内Ca2+荧光强度存在显著差异,外套膜外膜细胞内Ca2+荧光强度最高,内脏团上表皮细胞的最低(P<0.05);育珠三角帆蚌在相同Ca2+浓度的孵育水平下,外套膜细胞内Ca2+荧光强度比非育珠蚌都有增加的趋势,其中在1.25 mmol/L添加组中,两组外套膜内膜细胞内Ca2+荧光强度差异达到显著水平(P<0.05);不同Ca2+孵育水平对细胞内Ca2+荧光强度有显著影响(P<0.05),随着Ca2+在孵育液中浓度的升高,外套膜细胞内Ca2+荧光强度显著增强(P<0.05),表明外套膜是从外界吸收Ca2+的主要组织,蚌体珍珠的培育增强了其对Ca2+的沉积,并且水体Ca2+浓度在1.25～3.00 mmol/L有助于蚌体内Ca2+的贮藏,这对进一步开展淡水蚌类育珠学研究提供了基础理论,为珍珠培育的生产实践工作提供了依据。The Ca2＋metabolism is one of vital factors relative to growth of shell and pearl in freshwater mussel.In order to probe the Ca2＋ absorption and transportation in freshwater shellfish,Ca2＋ concentration of cells（under resting state）from different tissues and effect of Ca2＋ concentration of culture medium on Ca2＋deposits within the cells from outer mantle were researched by technology of laser scanning confocal microscopy in Hyriopsis cumingii.The total of healthy 10 H.cumingii with pearl and without pearl were selected,and then the cells from their mantle,gill,foot and epidermis of visceral mass were isolated.After short-term culture,the cells were incubated using DMEM with different Ca2＋ concentration（0,0.5,1.25,3 mmo1/L）and fluorescence probe of Fluo-3/AM for 1 h.The green fluorescence intensity was acquired to analyze the Ca2＋ levels in these cells.The results showed that there were significant differences among intracellular Ca2＋ fluorescent itensity of 5 kinds of cells（P〈0.05）：intracellular calcium ions fluorescent intensity of the outer mantle cells was highest（P〈0.05）,while intracellular Ca2＋fluorescent intensity of visceral mass epidermal cells was the lowest（P〈0.05）.The intracellular Ca2＋fluorescence intensity of mantle cells in mussel with pearl mussel had increasing trends compared to that in mussels without pearl under the same Ca2＋ level,and the difference was significant between the two groups of inner mantle with 1.25 mmol/L Ca2＋level（P〈0.05）.Influence of different Ca2＋ incubation levels on intracellular Ca2＋ fluorescence intensity were significant（P〈0.05）.With increasing Ca2＋ concentration in incubating medium,intracellular Ca2＋ fluorescence intensity of mantle cells was significantly enhanced（P〈0.05）.The present research indicated that mantle was main tissue on absorbing Ca2＋ from external water;pearl cultivation strengthened calcium deposition in body of H.cumingii;calcium ion concentration of 1.25-3 mmol/L in water w

关 键 词：三角帆蚌 细胞培养 激光共聚焦技术 Ca2+沉积 

分 类 号：Q952[生物学—动物学] S917[农业科学—水产科学]