内脏团插核术刺激对三角帆蚌血细胞的影响

EFFECT PEARL-NUCLEUS-INSERTING OPERATION IN VISCERAL MASS ON HAEMOCYTES IN HYRIOPSIS CUMINGII LEA

  • 摘要: 为了探讨三角帆蚌(Hyriopsis cumingii Lea)血细胞的类型及内脏团插核手术刺激对血细胞形态结构和数量的影响,研究利用相差显微镜、光学显微镜、透射电子显微镜和流式细胞仪对三角帆蚌血细胞进行了形态学研究。流式细胞术光散射图谱显示血细胞被分两类,一类为颗粒度高的大细胞,另外一类为颗粒度低的小细胞;相差显微镜观察显示,血细胞可分为胞体暗、折光性差和胞体明亮、折光性强的两类;Giemsa和H.E染色显示细胞分为胞质染色不均一、胞内颗粒明显和胞质染色均一、胞内颗粒不明显的两类;透射电镜超薄切片观察显示,颗粒明显的细胞胞质内线粒体、高尔基体等细胞器较丰富,颗粒不明显的细胞胞质内细胞器较少;负染结果表明血细胞主要分为表面不光滑、突起明显和细胞表面光滑、突起较不明显的两类。综合上述实验结果可见,三角帆蚌血细胞分为颗粒明显的细胞和颗粒不明显的透明细胞两大类。内脏团插核术刺激后,血细胞的形态和比例均发生显著变化。血细胞形态更多样,伪足状突起更明显,细胞内囊泡状物质增多,血细胞密度显著增高(PP<0.01)。研究表明,作为三角帆蚌免疫系统重要组成部分的血细胞,在插核手术后,其类型、形态结构和数量均产生明显变化,这是机体对外界刺激产生的免疫防御反应,其中颗粒细胞担负着主要的免疫功能。

     

    Abstract: In order to investigate the haemocyte types and the effect pearl-nucleus-inserting operation in visceral mass on haemocytes in Hyriopsis cumingii Lea, haemocyte morphology were studied by phase contrast microscope, optical microscope, transmission electron microscope and flow cytometry FCM. The present studies showed that two cell populations were found in haemocytes: large cell with high granularity and small cell with low granularity by flow cytometry FCM on light scanttering pattern. Two distinct cell types were identified based on phase contrast microscope: one type of cell was dark and dioptric aberration, while the other was bright and dioptric strong. By Giemsa and H.E staining, cytoplasmic staining were heterogeneous and internal particles were obvious in one type of cell, while cytoplastic staining were homogeneous and internal particles were inexistent in the other type of cell. By transmission electron microscope, we found that the mitochondria, Golgi apparatus organelles were rich and internal particles were obvious in one type of cells, and contrary to the another cells. The results showed that the surface was unsmooth and pseudopodium was obvious in one type of haemocyte, while smooth and unobvious in another by negative staining. All of these indices indicated that the haemocytes were divided into two types: granulocyte and hyalinocyte in Hyriopsis cumingii. When stimulated by pearl-nucleus-inserting operation in visceral mass, the proportion and morphology of haemocytes underwent manifest changes: more haemocyte morphologies, more protuberances, more vesicle-like material, significant increase for density of haemocytes and quantity of granulocytes (PP<0.01) post-operation. The present research indicated that there were obvious changes in the morphology, structures and proportion of populations of haemocyte by pearl-nucleus-inserting operation in visceral mass, which implied immune defense was enhanced after the operation, and the granulocyte played an important role in immune system.

     

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