泥蚶血细胞的形态结构特征及部分免疫功能  

Morphological,structural characteristics and phagocytic and enzymatic activities of haemocytes in blood clam Tegillarca granosa

作　　者：朱泽闻[1] 徐立蒲 吴信忠[1] 张振国 吴刘记[1] 娄绘芳[1] 

机构地区：[1]浙江大学动物科学学院,浙江杭州310029 [2]北京市水产技术推广总站,北京100025

出　　处：《水产学报》2011年第10期1494-1504,共11页Journal of Fisheries of China

基　　金：国家自然科学基金项目(39970581;30170741);浙江省自然科学基金项目(20080123)

摘　　要：利用光镜、透射电镜、扫描电镜及流式细胞仪技术对泥蚶血细胞形态结构以及吞噬活性进行了研究。根据细胞大小、形态结构、颗粒特点,光镜下可将血细胞区分为红色颗粒细胞(占89.67%)、嗜碱性颗粒细胞(占7.05%)、透明细胞(占3.28%)3种主要类型。健康泥蚶血淋巴中血细胞平均密度为(3.29±0.82)×106/mL。透射电镜下观察到颗粒细胞Ⅰ型、颗粒细胞Ⅱ型、无颗粒细胞;扫描电镜观察下,细胞表面光滑,部分细胞表面有小的突起物。用流式细胞仪检测出P1(13.13%)亚群和P2(78.83%)两个细胞亚群以及没有固定形态的细胞(近8%)。利用APIZYM试剂盒对健康泥蚶血细胞及血清中的19种酶进行检测,在血细胞中检测到11种酶,在血清中检测到了13种酶,经酵母聚糖刺激后,碱性磷酸酶、酸性磷酸酶、类脂脂酶、N-乙酰葡萄糖胺酶在血细胞和血清中的水平大幅升高。体外吞噬实验观察显示,红色颗粒细胞对酵母聚糖具有明显的吞噬能力,嗜碱性颗粒细胞和透明细胞吞噬作用不显著;在经嗜水气单胞杆菌刺激后,嗜碱性颗粒细胞出现大量聚集现象,透明细胞能吸附颗粒物。吞噬活性研究表明,温度、盐度对泥蚶血细胞的吞噬活性有显著的影响,在泥蚶适宜生长温度(13～30℃)区间和最适盐度(20.0～26.2)×10-12区间,血细胞吞噬活性达到最高值。Light,electron microscopic and flow cytometer studies were conducted to characterize morphology and structure and phagocytosis of haemocytes in blood clam Tegillarca granosa.Based on cell size,and microstructure and granular types of cell,blood clam haemocytes were divided into red granulocyte（89.67%）,basophil granulocyte（7.05%）and hyalinocyte（3.28%）respectively under light microscopy.The haemocyte densities in healthy blood clam haemolymph were assessed（3.29±0.82）×106 cell/mL.Three kinds of haemocytes were respectively recognized by transmission electron microscope：granulocyte type Ⅰ,granulocyte type Ⅱ and agranulocyte.It was observed under scanning electron microscopy that haemocytes appear smooth-surface and some of them have knot-like projections in cell surface.APIZYM assays on enzymatic activities of 19 enzymes in haemolymph were performed and,of these,13 and 10 enzymes were detected respectively in serum and cell.There were marked increases in the levels of ALP,ACP,ELIP（C8）and AGLA in serum and haemocytes after being stimulated by zymosan.Observation of phagocytosis in vitro revealed that there was phagocytic response in red granulocyte to zymosan and there was no significant phagocytic response in basophil granulocyte and hyalinocyte.It was observed that basophilic granulocyte accumulation and hyalinocyte adsorption after being stimulated by Aeromonas hydrophila.The study of environmental stress factors on phagocytosis activity revealed that the temperature and salinity have significant influence on phagocytosis activity.The phagocytosis percentage of blood clam reached the highest values with optimum growth temperature（13-30 ℃）and salinity（20.0-26.2）×10-12.

关 键 词：泥蚶 血细胞 显微结构 酶活性 吞噬活性 颗粒细胞 透明细胞 

分 类 号：Q178.1[生物学—水生生物学] S917.4[生物学—普通生物学]