| 红螯螯虾血细胞分类、结构与免疫特性研究 |
| Classification, Structure and Immunological Characterization of Haemocytes in Red Claw Crayfish, Cherax quadricarinatus |
| 投稿时间：2017-09-30 修订日期：2019-09-17 |
| DOI：10.15928/j.1674-3075.2019.05.013 |
| 中文关键词:红螯螯虾 流式细胞术 血细胞 免疫力 |
| 英文关键词:Cherax quadricarinatus flow cytometry haemocyte immunity |
| 基金项目:中国热带农业科学院基本科研业务费专项资金（1630052016011，1630052019013）；海南省重点研发计划项目（ZDYF2016084） |
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| 中文摘要: |
| 血细胞在甲壳动物免疫过程中起重要作用，探讨红螯螯虾（Cherax quadricarinatus）不同类型血细胞结构与免疫特性有助于其病害防治的进一步深入研究。应用流式细胞术（FCM），根据前向角散射光（FSC）和侧向角散射光（SSC）的强度差异对血细胞进行分类，利用特异性荧光染料进行标记，分析血细胞总数（THC）、线粒体数量、溶酶体数量、吞噬活力、活性氧（ROS）含量、一氧化氮（NO）含量和非特异性酯酶活力。结果显示，应用FCM可以区分透明细胞（HC）、半颗粒细胞（SGC）和颗粒细胞（GC）三类血细胞，其占比分别为9.82%、61.11%和25.24%；红螯螯虾平均血细胞总数（THC）为（8.43±0.87）×106个/mL；GC含有最多数量的线粒体和溶酶体，HC中含量最少；HC、SGC和GC的吞噬率分别为2.54%、14.45%和6.98%，SGC吞噬活力最强，HC最弱；HC、SGC和GC的活性氧（ROS）含量分别为7.80、45.95和134.69 AU，GC的ROS含量最高，HC最低；HC、SGC和GC的一氧化氮（NO）含量分别为8.20、79.78和344.31 AU，GC的NO含量最高，HC最低；HC、SGC和GC的非特异性酯酶活力分别为86.59、121.84和236.91AU，GC的酯酶活力最高，HC最低。研究表明，红螯螯虾三类血细胞在形态结构、数量及免疫特性上均存在差异，SGC的数量最多、吞噬活力最强，而GC含有最多与能量供应、免疫防御相关的细胞器，并拥有最强的氧化活力和酯酶活力，表明GC和SGC是红螯螯虾免疫防御过程中发挥主要作用的血细胞类型。 |
| 英文摘要: |
| Haemocytes play vital role in the immunological response of crustaceans and our study of the structure and immune characteristics of haemocytes in red claw crayfish (Cherax quadricarinatus) supports disease control. In this study, we classified haemocytes by flow cytometry (FCM) based on difference in the intensity of forward scattered light (FSC) and side scattered light (SSC). Total haemocyte count (THC), mitochondria number, lysosome number, phagocytosis, production of reactive oxygen species (ROS) and NO, and non-specific esterase activity of haemocytes were determined using specific fluorescent dyes for labeling. The study provides basic data for immunology and disease control in red claw crayfish. Three types of haemocytes were identified by FCM： hyaline cells (HC, 9.82%), semigranular cells (SGC, 61.11%) and granular cells (GC, 25.24%). The mean THC was （8.43±0.87）x106 cells/mL in C. quadricarinatus. The masses of mitochondria and lysosomes were highest in GC and lowest in HC. Phagocytic rates of HC, SGC and GC were, respectively, 2.54%, 14.45% and 6.98%. ROS production, NO production and non-specific esterase activity of HC, SGC and GC were, respectively: 7.80, 45.95 and 134.69 AU; 8.20, 79.78 and 344.31AU; 86.59, 121.84 and 236.91AU. In summary, the three haemocyte types of C. quadricarinatus displayed variation in morphologic structure, amount and immunological characteristics. The number and phagocytic activity of SGC were highest, while GC contained the most organelles in relation to energy supply and immune response, and has the highest oxidative and esterase activities. These findings indicate that GC and SGC are the primary haemocyte types involved in the immunological response of C. quadricarinatus. |
| 冼健安,张秀霞,潘训彬,鲁耀鹏,李军涛,段瑞军,郑佩华,王冬梅.2019.红螯螯虾血细胞分类、结构与免疫特性研究[J].水生态学杂志,40(5):84-90. |
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