Histological observation and innate immune barrier study of head kidney of Pelteobagrus vachelli
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摘要: 为探明瓦氏黄颡鱼 (Pelteobagrus vachelli) 头肾组织结构和先天性免疫屏障，该研究用苏木素-伊红 (HE) 染色法和改良James染色法对瓦氏黄颡鱼头肾的显微结构进行了观察，并使用台盼蓝活体染色法对头肾免疫屏障部位进行了定位。结果显示，瓦氏黄颡鱼头肾由被膜、淋巴细胞中心区、外周区、前肾间组织、网状纤维和胶原纤维组成，胶原纤维仅在血管外周可见；台盼蓝主要分布于血窦周围的内皮细胞和一些游离的巨噬细胞中，且随时间延长呈递增趋势。研究表明血窦周围的内皮细胞和巨噬细胞组成了瓦氏黄颡鱼的头肾先天性免疫屏障。Abstract: To investigate the histological structure and immune function of head kidney of Pelteobagrus vachelli, we used hematoxylin-eosin (HE) staining and modified James staining to observe its microscopic structure, and applied trypan blue vital staining to locate its immune barrier site. The results show that the head kidney of P. wallichii consisted of capsule, lymphocyte central area, peripheral area, anterior interrenal tissue, reticular fibers and collagen fibers, and collagen fibers were only visible at the periphery of blood vessels. Trypan blue was mainly distributed in endothelial cells around sinusoids and some free melanin macrophages, showing an increasing trend with time. Results suggest that endothelial cells of red pulps and free macrophages consist of the innate immune barrier of head kidney.
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Key words:
- Pelteobagrus vachelli /
- Head and kidney /
- Tissue structure /
- Immune function
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图 1 瓦氏黄颡鱼头肾解剖示意图
a.头肾的解剖位置；b.头肾的形态
Figure 1. Anatomical diagram of head kidney of P. vachelli
a. Anatomical location of head kidney; b. Anatomical structure of head kidney
图 2 瓦氏黄颡鱼头肾的组织结构HE染色观察
a.被膜 (→)，血窦 (▲)；b. 淋巴细胞聚集中央区 (○) 与外周区 (□)；c. 肾间组织 (★)；d. 内皮细胞 (→)和黑色素巨噬细胞中心 (▲)
Figure 2. HE Staining observation of head and kidney of P. vachelli
a. Capsule (→), blood sinus (▲); b. Lymphocyte aggregation central region (○) and peripheral region (□); c. Anterior inter-renal tissue (★); d. Endothelial cells (→) and melanin macrophage centers (▲)
图 3 瓦氏黄颡鱼正常头肾组织切片改良James染色观察
a．网状纤维覆盖于整个头肾 (→)；b. 分布于头肾被膜的网状纤维 (→)；c. 小静脉血管内侧及血管周围的网状纤维 (→)；d. 大动脉血管周围的胶原纤维 (→)；e. 黑色素巨噬细胞外层的网状纤维 (→)；f. 肾间组织内的网状纤维 (→)
Figure 3. Modified James staining observation of normal head kidney tissue sections of P. vachelli
a. Reticular fibers covered the entire head kidney (→); b. Reticular fibers (→) distributed in the cephalic renal capsule; c. Reticular fibers (→) inside and around venules; d. Collagen fibers around the aorta (→); e. Reticular fibers (→) on the outer layer of melanin macrophages; f. Reticular fibers (→) in anterior renal interstitials
图 4 注射台盼蓝不同时间点瓦氏黄颡鱼体颜色变化
a.正常体色，未注射台盼蓝；b. 注射第24小时；c. 注射第48小时；d. 注射第72小时
Figure 4. Changes of fish color at different time points of injection trypan blue in P. vachelli
a. Normal body color without trypan blue injection; b. 24 h after injection; c. 48 h after injection; d. 72 h after injection
图 5 瓦氏黄颡鱼头肾中台盼蓝的分布 (未染色)
a. 0 h未注射台盼蓝，正常头肾；b. 4 h台盼蓝开始出现在一些细胞里面，呈点状或小聚集团状 (→)；c. 第8 h台盼蓝开始增多(→)；d. 第24 h台盼蓝数量增多较明显，可明显看见小聚集团面积增大(→)；e. 第48 h台盼蓝增加特别明显，可看见大面积台盼蓝聚集团(→)；f. 第72 h台盼蓝数量继续增多(→)
Figure 5. Distribution of trypan blue in head kidney of P. vachelli (Unstained)
a. Trypan blue was not injected at the 0th hour, normal head and kidney; b. Trypan blue began to appear in some cells at the 4th hour, showing punctate or small aggregates (→); c. Trypan blue began to increase at the 8th hour (→); d. The number of Trypan blue increased more obviously at the 24th hour, and the area of small aggregates could be clearly seen (→); e. The increase of Trypan blue at the 48th hour was particularly obvious, and large area of Trypan blue aggregates could be seen (→); f. The number of trypan blue continued to increase at the 72th hour (→).
图 6 瓦氏黄颡鱼头肾中第0至第72小时台盼蓝含量变化趋势
*. 0.01<P<0.05; **. P<0.01
Figure 6. Variation trend of trypan blue content in head kidney of P. vachelli from 0th to 72nd hour
图 7 瓦氏黄颡鱼台盼蓝的分布部位
a. 第48小时头肾，he染色，示台盼蓝(→)；b. a中同一区域头肾未染色，示台盼蓝(→)；c. 第72 小时头肾，he染色，示台盼蓝(→)；d、c中同一区域头肾未染色，示台盼蓝(→)；e. 网状内皮细胞(→)；f. 血窦中游离的巨噬细胞(→)
Figure 7. Distribution of trypan blue in P. vachelli
a. 48 h head kidney, HE staining, showing trypan blue (→); b. The same area of A, unstained, showing trypan blue (→); c. 72th hour head kidney, HE staining, showing trypan blue (→); d. The same area of C, unstained, showing trypan blue (→); e. Reticular endothelial cells (→); f. Free macrophages in blood sinus (→)
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