川陕哲罗鲑鳃组织的显微与超微结构观察

Microstructure and ultrastructure observation of gill tissue in Hucho bleekeri

  • 摘要:目的】观察分析川陕哲罗鲑鳃组织的显微与超微结构,丰富其基础生物学资料,为揭示鱼类鳃组织结构特征与其生活习性的关联提供参考依据。【方法】以人工繁育的川陕哲罗鲑F1代为研究对象,采集鳃组织样品分别用4%多聚甲醛和3%戊二醛保存固定,经石蜡包埋切片后依次进行苏木素—伊红(HE)染色、阿利辛蓝—过碘酸雪夫(AB-PAS)染色、Masson染色及天狼星红染色,光学显微镜下观察鳃组织学形态、黏液细胞类型、胶原纤维分布与分型;同时利用扫描电子显微镜和透射电子显微镜观察鳃组织的超微结构。【结果】川陕哲罗鲑有4对鳃,鳃耙呈乳白色针形,排列稀疏(12~14根);鳃丝呈鲜红色,长条状,由基部向前端延伸的过程中逐渐变细小,其末端呈游离状态。鳃弓主要由软骨组织构成,被覆复层上皮细胞;鳃丝表面分布有黏液细胞和泌氯细胞,中央有软骨细胞支撑;鳃小片由呼吸上皮及柱状上皮组成,相邻2个柱细胞的胞体延长即围成毛细血管腔,腔内可见红细胞。AB-PAS染色结果显示,鳃组织中的黏液细胞以分泌中性黏多糖为主(呈PAS阳性),伴有少量酸性黏多糖(呈AB阳性)。Masson染色发现,鳃组织胶原纤维分布均匀,胶原纤维组织面积占比为0.21%~0.31%(平均为0.27%),未见纤维化等典型病变。天狼星红染色结果显示,I型胶原纤维的占比为80.00%~90.49%,Ⅲ型胶原纤维的占比为9.51%~19.20%,I型胶原纤维/Ⅲ型胶原纤维的比例为4.21~9.51。扫描电子显微镜观察发现,鳃耙表面凹凸不平且有孔洞,鳃弓及鳃丝上皮细胞具有密集的环形微嵴,呈指纹状;鳃丝表面与鳃小片均有黏液,但鳃丝表面的孔洞数量多于鳃小片。透射电子显微镜观察发现,鳃小片主要由扁平上皮细胞、柱细胞、泌氯细胞、巨噬细胞、红细胞、内皮细胞及血管腔中的中性粒细胞等组成,其中,泌氯细胞的细胞核呈长椭圆形,核直径长边约8.3 μm、短边约5.6 μm,细胞质富含线粒体和粗面内质网。【结论】川陕哲罗鲑的鳃组织结构与其肉食性及高海拔、低水温、急流淡水生境高度适应;且川陕哲罗鲑鳃组织含有巨噬细胞及中性粒细胞等免疫细胞,构成其特有的黏膜免疫屏障,在先天免疫系统中发挥重要作用。

     

    Abstract:Objective】This study aimed to observe and analyze microstructure and ultrastructure of gill tissue in Hucho bleekeri and enrich its basic biological data, providing reference for elucidating the association between gill structure characteristics and life habits in fishes.【Method】The F1 generation of artificially bred Hucho bleekeri was used as the research subject. Gill tissue samples were collected and fixed in 4% paraformaldehyde and 3% glutaraldehyde respectively. After paraffin embedding and sectioning, the samples were sequentially subjected to hematoxylin-eosin (HE) staining, Alcian blue-periodic acid Schiff (AB-PAS) staining, Masson staining, and Sirius red staining. The histological morphology, mucous cell types, and distribution and classification of collagen fibers were observed under a light microscope. Meanwhile, the ultrastructure of gill tissue was examined using scanning electron microscopy and transmission electron microscopy.【ResultHucho bleekeri possessed four pairs of gills. The gill rakers were milky-white, needle-shaped, and sparsely arranged (12-14 rakers). The gill filaments were bright red and elongated, gradually tapering from the base toward the distal end, with the terminal portion remaining free. The gill arch was primarily composed of cartilaginous tissues and covered by stratified epithelial cells. The gill filament surface was distributed with mucous cells and chloride-secreting cells, with chondrocytes providing central support. The gill lamella consisted of respiratory epithelium and pillar epithelium, and the extended cell bodies of two adjacent pillar cells enclosed the capillary lumen, within which erythrocytes were visible. AB-PAS staining results indicated that mucous cells in gill tissue predominantly secreted neutral mucopolysaccharides (PAS-positive), accompanied by a small amount of acidic mucopolysaccharides (AB-positive). Masson staining revealed that collagen fibers were evenly distributed in gill tissue, with a collagen fiber tissue area proportion of 0.21%-0.31% (the mean was 0.27%), and no typical lesions such as fibrosis were observed. Sirius red staining results showed that type I collagen fibers accounted for 80.00%-90.49%, type Ⅲ collagen fibers accounted for 9.51%-19.20%, and the ratio of type I/type Ⅲ collagen fibers ranged from 4.21 to 9.51. Scanning electron microscopy observation revealed that the gill raker surface was uneven and porous; the epithelial cells of gill arches and filaments possessed dense circular microridges arranged in a fingerprintlike pattern; mucus was found on both gill filament surface and gill lamellae, but the number of pores on gill filament surface exceeded that on the gill lamellae. Transmission electron microscopy observation revealed that the gill lamella was mainly composed of flattened epithelial cells, pillar cells, chloride-secreting cells, macrophages, erythrocytes, endothelial cells, and neutrophils in the vascular lumen. Among these, the chloride-secreting cells had an elongated oval nucleus, with the long diameter at 8.3 μm and short diameter at 5.6 μm approximately, and the cytoplasm had abundant mitochondria and rough endoplasmic reticulum.【Conclusion】The gill tissue structure in Hucho bleekeri is highly adapted to its carnivorous nature and the habitat of high altitude, low temperature, and fast-flowing freshwater. Moreover, the gill tissue of Hucho bleekeri have immune cells such as macrophages and neutrophils, constituting a specific mucosal immune barrier that plays an important role in innate immune system.

     

/

返回文章
返回