4种黄河珍稀鱼类外周血细胞显微与超微结构研究

Microscopic and ultrastructural study on peripheral blood cells of 4 rare fish species in Yellow River

  • 摘要: 【目的】 解析花斑裸鲤(Gymnocypris eckloni)、极边扁咽齿鱼(Platypharodon extremus)、黄河裸裂尻鱼(Schizopygopsis pylzovi)和厚唇裸重唇鱼(Gymnodiptychus pachycheilus)外周血细胞显微与超微结构,丰富4种黄河珍稀鱼类血液学参数,为黄河珍稀鱼类资源保护提供生理学资料。【方法】 使用ArcGIS Desktop 10.5绘制采样点示意图,利用刺网和地笼进行捕捞。4种黄河珍稀鱼类经MS-222麻醉后,采用经肝素钠抗凝处理的1 mL注射器自尾静脉采血,并制备显微与超微结构样本。选择染色良好的外周血涂片,统计各类细胞占比,并使用ImageJ测量不同类型血细胞长径和短径及细胞核长径和短径。【结果】 细胞显微结构和超微结构观察结果显示,除成熟红细胞外,在4种黄河珍稀鱼类外周血中还观察到未成熟红细胞和分裂期红细胞,4种黄河珍稀鱼类外周血细胞显微结构均未观察到嗜碱性粒细胞,但在超微结构中观察到少量嗜碱性粒细胞。厚唇裸重唇鱼嗜酸性粒细胞中的特殊颗粒由致密的晶体核心和基质组成,区别于其他3种鱼类。4种黄河珍稀鱼类外周血中粒细胞和单核细胞表面均分布伪足,且单核细胞胞质内线粒体和粗面内质网丰富,在厚唇裸重唇鱼单核细胞中还观察到初级溶酶体和次级溶酶体。细胞占比统计与大小测量结果显示,4种黄河珍稀鱼类外周血中淋巴细胞占比均最高,其次为嗜中性粒细胞,嗜酸性粒细胞和单核细胞占比较低。4种黄河珍稀鱼类外周血中红细胞长径无显著差异(P>0.05),花斑裸鲤红细胞短径显著大于其他3种鱼类(P<0.05,下同),厚唇裸重唇鱼红细胞胞核长径和短径显著大于其他3种鱼类,极边扁咽齿鱼嗜酸性粒细胞长径和短径显著大于厚唇裸重唇鱼,细胞核长径和短径显著大于黄河裸裂尻鱼,花斑裸鲤小淋巴细胞短径显著大于极边扁咽齿鱼,细胞核长径和短径显著大于黄河裸裂尻鱼。【结论】 4种黄河珍稀鱼类外周血细胞组成相似,显微结构均可见红细胞、血栓细胞、嗜中性粒细胞、嗜酸性粒细胞、单核细胞和淋巴细胞,超微结构还可见嗜碱性粒细胞。4种黄河珍稀鱼类外周血成熟红细胞体积相近,可能与其生活在相同水域环境有关。4种黄河珍稀鱼类外周血淋巴细胞占比最高,其次为嗜中性粒细胞,丰富的淋巴细胞可能与其形态适应性相关。

     

    Abstract: 【Objective】 The aim of this study was to analyze the microscopic and ultrastructural characteristics of peripheral blood cells of Gymnocypris eckloni, Platypharodon extremus, Schizopygopsis pylzovi and Gymnodiptychus pachycheilus, enrich the hematological parameters of the 4 rare fish species in Yellow River, and provide physiological data for the protection of rare fish resources in Yellow River. 【Method】 ArcGIS Desktop 10.5 was used to draw a schematic diagram of the sampling sites, gill nets and ground traps were used for fishing. After the 4 rare fish species in Yellow River were anesthetized with MS-222, blood was collected from the tail vein with a 1 mL syringe treated with sodium heparin anticoagulation, and microstructural and ultrastructural samples were prepared. Well-stained peripheral blood smears were selected, and the proportion of each type of cells was counted. ImageJ was used to measure the long and short diameters of different types of blood cells and the long and short diameters of the cell nucleus. 【Result】 The results of cell microstructure and ultrastructure observation showed that in addition to mature erythrocytes, immature erythrocytes and mitotic erythrocytes were also observed in the peripheral blood of the 4 rare fish species in Yellow River. No basophils were observed in the microstructure of peripheral blood cells of the 4 rare fish species in Yellow River, but a small number of basophils were observed in the ultrastructure. The special granules in the eosinophils of Gymnodiptychus pachycheilus were composed of dense crystal core and matrix, which was different from the other 3 fish species. In the peripheral blood of the 4 rare fish species in Yellow River, the surfaces of granulocytes and monocytes were all distributed with pseudopodia, and the mitochondria and rough endoplasmic reticulum in the cytoplasm of monocytes were well developed. Primary lysosomes and secondary lysosomes were also observed in the monocytes of Gymnodiptychus pachycheilus. The results of cell proportion statistics and size measurement showed that the proportion of lymphocytes in the peripheral blood of the 4 rare fish species in Yellow River was the highest, followed by neutrophils, and the proportion of eosinophils and monocytes was relatively low. There was no significant difference in the long diameter of erythrocytes in the peripheral blood of the 4 rare fish species in Yellow River(P>0.05). The short diameter of erythrocytes in Gymnocypris eckloni was significantly larger than that in the other 3 fish species(P<0.05, the same below). The long and short diameters of the erythrocyte nucleus of Gymnodiptychus pachycheilus were significantly larger than those in the other 3 fish species. The long and short diameters of eosinophils in Platypharodon extremus were significantly larger than those in Gymnodiptychus pachycheilus, and the long and short diameters of the nucleus were significantly larger than those in Schizopygopsis pylzovi. The short diameter of small lymphocytes in Gymnocypris eckloni was significantly larger than that in Platypharodon extremus, and the long and short diameters of the nucleus were significantly larger than those in Schizopygopsis pylzovi. 【Conclusion】 The peripheral blood cell composition of the 4 rare fish species in Yellow River is similar. Erythrocytes, thrombocytes, neutrophils, eosinophils, monocytes and lymphocytes can be seen in the microstructure, and basophils can also be seen in the ultrastructure. The mature erythrocytes of the 4 rare fish species in Yellow River are similar in size, which may be related to the fact that they live in the same water environment. Lymphocytes account for the highest proportion in the peripheral blood of the 4 rare fish species in Yellow River, followed by neutrophils. The abundant lymphocytes may be related to their morphological adaptability.

     

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