海菊蛤血细胞类型和吞噬功能的研究
周可欣(2002—), 女, 广东省汕头市人, 本科生。email: 3072997544@qq.com |
Copy editor: 殷波 , YIN Bo
收稿日期: 2024-05-25
修回日期: 2024-07-22
网络出版日期: 2024-08-14
基金资助
广东省教育厅海洋贝类生态养殖与病害防控创新团队项目(2021KCXTD057)
惠州学院教授、博士启动项目(15604220001)
Morphological classification and phagocytosis of haemocytes in Spondylus sp.
Copy editor: YIN Bo
Received date: 2024-05-25
Revised date: 2024-07-22
Online published: 2024-08-14
Supported by
Guangdong Provincial Department of Education Marine Shellfish Ecological Aquaculture and Disease Prevention and Control Innovation Team Project(2021KCXTD057)
Professors and doctors of Huizhou University started the project(15604220001)
在贝类免疫防御体系中, 血细胞作为其主要的免疫效应细胞, 起着非常重要的作用。文章通过血细胞计数、Giemsa染色、透射电镜、扫描电镜和荧光吞噬的方法研究了海菊蛤血细胞的形态与类型及吞噬功能。海菊蛤的血细胞密度为3.296 × 106cells·mL-1, 血细胞在大小和胞质内是否含有颗粒方面存在显著差异, 基于这些差异, 将其细分为透明细胞(占9.74%)、颗粒细胞(占58.14%)、无颗粒细胞(占27.50%)和淋巴样细胞(占4.62%) 4种类型。透射电镜下可以观察到透明细胞、颗粒细胞、无颗粒细胞和淋巴样细胞。扫描电镜观察, 可以明确区分出4种不同形态的细胞, 它们分别是椭圆细胞、圆形细胞、梭形细胞、核形(锥形)细胞。利用标记了绿色荧光蛋白(green fluorescent protein, GFP)的大肠杆菌BL21进行血细胞吞噬试验, 结果显示, 海菊蛤血细胞的吞噬作用主要依赖于颗粒细胞来完成。
周可欣 , 甄雯婧 , 张文文 , 王江勇 . 海菊蛤血细胞类型和吞噬功能的研究[J]. 热带海洋学报, 2025 , 44(2) : 84 -91 . DOI: 10.11978/2024108
Blood cells, as the main immune effector cells, play a very important role in the immune defense system of shellfish. The morphology, type and phagocytosis function of hemocytes of Spondylus sp. were studied by means of blood cell count, Giemsa staining, transmission electron microscopy, scanning electron microscopy and fluorescence phagocytosis. The results showed that the blood cell density of the Spondylus sp. was 3.296 × 106 cells·mL-1, and the blood cells differ significantly in size and with regard to whether they contain particles within the cytoplasm. Based on these differences, they are subdivided them into four types: hyalinocyte (9.74%), granulocyte (58.14%), agranular haemocyte (27.50%) and lymphoid (4.62%). Hyalinocytes, agranular haemocytes, granulocytes and lymphoids could be observed under transmission electron microscope. Under the observation of scanning electron microscope, four different types of cells can be clearly distinguished, which are oval cells, round cells, spindle cells, nucleated (conical) cells. A phagocytosis experiment was carried out with Escherichia coli BL21 labeled with the green fluorescent protein (GFP), and the results showed that the phagocytosis of the blood cells from Spondylus sp. was mainly dependent on granulocytes.
图1 海菊蛤血细胞光学显微镜观察(1000 ×)G: 颗粒细胞; AG: 无颗粒细胞; HC: 透明细胞; LY: 淋巴样细胞 Fig. 1 Optical microscope observation of haemocytes in Spondylus sp. (1000 ×). G: granulocyte; AG: agranular haemocyte; HC: hyalinocyte; LY: lymphoid haemocyte |
图3 海菊蛤血细胞透射电子显微镜观察(4000 ×)a, b. 颗粒细胞; c, d. 透明细胞; e, f. 淋巴样细胞; f. 无颗粒细胞。N: 细胞核; RER: 粗面内质网; LYS: 溶酶体; M: 线粒体; V: 液泡; LY: 淋巴样细胞; AG: 无颗粒细胞 Fig. 3 Morphology of haemocytes in Spondylus sp. by transmission electron microscope (4000 ×). a, b: granulocyte; c, d: hyalinocyte; e, f: lymphoid haemocyte; f: agranulocyte. N: nucleus; RER: rough endoplasmic reticulum; LYS: lysosome; M: mitochondria; V: vacuole; LY: lymphoid haemocyte; AG: agranular haemocyte |
图4 海菊蛤血细胞的形态学-扫描电子显微镜观察a. 海菊蛤细胞(1500 ×); b. 圆形细胞(10000 ×); c. 椭圆细胞(10000 ×); d. 梭形细胞(10000 ×); e. 梭形细胞(7000 ×); f. 核形(锥形)细胞(8000 ×) Fig. 4 Morphology of haemocytes in Spondylus sp. by scanning electron microscope. a: spondylidae cells (1500 ×); b: round cells (10000 ×); c: elliptical cells (10000 ×); d: spindle cells (10000 ×); e: spindle cells (7000 ×); f: nucleated (conical) cells (8000 ×) |
图5 海菊蛤血细胞的吞噬实验-荧光显微镜观察a, d. 明场/海菊蛤血细胞(400 ×); b, e. GFP/大肠杆菌BL21 (400 ×); c, f. Merge (400 ×)。G: 颗粒细胞; HC: 透明细胞 Fig. 5 Phagocytosis experiment and fluorescence microscope observation of haemocytes in Spondylus sp.. a, d: Bright field/Spondylus sp. haemocyte (400 ×); b, e: GFP/Escherichia coli BL21 (400 ×); c, f: Merge (400 ×). G: granulocyte; HC: hyalinocyte |
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