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一种甲壳动物血细胞分化因子性质的初步分析
李鸿钰,杨丰,李钫
0
(自然资源部第三海洋研究所,福建 厦门 361005)
摘要:
血细胞是甲壳动物的免疫细胞,负责抵御和清除入侵的病原。虽然血细胞在甲壳动物的整个生命周期中都需要不断更新,但人们对甲壳动物造血机制的了解却十分有限。前期研究发现利用红螯螯虾的肌肉提取液可以诱导体外培养的造血组织细胞定向分化为颗粒细胞,但其中起到诱导分化作用的因子尚未明确。本研究对该未知血细胞分化因子的基本性质进行了分析。首先,我们制备了红螯螯虾(Cherax quadricarinatus)、中华绒螯蟹(Erinocheir sinensis)、东亚飞蝗(Locusta migratoria manilensis)、红石斑鱼(Epinephelus goreensis)、凡纳滨对虾(Litopenaeus vannamei)和疣荔枝螺(Reishia clavigera)的肌肉提取液,测试其诱导螯虾造血组织细胞分化的能力。结果表明,以上各种动物的肌肉提取液均可诱导原代培养的螯虾造血组织细胞向颗粒细胞分化,说明该血细胞分化因子普遍存在于这些动物的肌肉组织中。进一步我们以螯虾肌肉提取液为实验对象,分析了血细胞分化因子的热稳定性、溶解性、带电性、对蛋白酶的耐受性以及分子量大小。结果发现该血细胞分化因子易溶于甲醇,呈电中性,具有良好的热稳定性,且可耐受蛋白酶处理,分子量小于1 kDa。因此,螯虾肌肉提取液中可诱导造血组织细胞分化为颗粒细胞的血细胞分化因子是一种广泛存在的小分子类物质。本研究结果有助于后续对该因子的分离纯化和鉴定。
关键词:  海洋生物学  血细胞分化因子  造血组织细胞  肌肉提取液  甲壳动物
DOI:10.3969/J.ISSN.2095-4972.20220314002
基金项目:国家自然科学基金(31672675)
Preliminary analysis of the characterization of a crustacean hemocyte differentiation factor
LI Hongyu,YANG Feng,LI Fang
(Third Institute of Oceanography, MNR, Xiamen 361005, China)
Abstract:
As the primary immune cells, hemocytes are produced throughout the life cycle of crustacean. However, our understanding of crustacean hematopoiesis is limited. Recently, we found that the muscle extract of Cherax quadricarinatus can induce the differentiation of hematopoietic tissue (HPT) cell into granular cells in vitro. The exact factor that induces GC differentiation remains unknown. In this study, the basic properties of the unknown hemocytes differentiation factor were analyzed. We prepared muscle extracts from C. quadricarinatus, Eriocheir sinensis, Locusta migratoria manilensis, Epinephelus goreensis, Litopenaeus vannamei and Reishia clavigera. Their ability to induce HPT cells differentiation was tested. Results showed that all the muscle extracts could induce the differentiation of HPT cells along the GCs lineage, indicating that the hemocyte differentiation factor exists in all tested animals. Further, we analyzed the thermal stability, solubility, electrification, resistance to protease and molecular weight of the hemocytes differentiation factor in the muscle extract of C. quadricarinatus. We found that the target factor was a small molecule (< 1 kDa), which was methanol-soluble, electrically neutral, thermal stable and protease-resistant. Therefore, the target factor is a widely existing small molecule. Thus, results of this study should facilitate the purification and identification of this factor.
Key words:  marine biology  hemocytes differentiation factor  hematopoietic tissue cell  muscle extract  crustaceans

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