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大西洋鲑幼鱼肝脏的光镜、透射电镜和扫描电镜研究,特别关注肝血窦

Liver of juvenile Atlantic salmon, Salmo salar L.: a light, transmission, and scanning electron microscopic study, with special reference to the sinusoid.

作者信息

Speilberg L, Evensen O, Nafstad P

机构信息

Department of Pathology, Central Veterinary Laboratory, Oslo, Norway.

出版信息

Anat Rec. 1994 Nov;240(3):291-307. doi: 10.1002/ar.1092400302.

Abstract

BACKGROUND

This report provides a detailed description of sinusoidal and perisinusoidal structures in the normal liver of the juvenile Atlantic salmon (Salmo salar L.), a teleost species.

METHODS

The liver was studied by light, transmission, and scanning electron microscopy, and organ specimens were sampled after retrograde, whole-body perfusion through the dorsal aorta using 3% glutaraldehyde. Detailed characterization of perisinusoidal stellate cells also included immunohistochemical staining for desmin and evaluation of autofluorescence of the same cells upon excitation in ultraviolet (UV) light.

RESULTS

The sinusoid is lined by one cell type only: the endothelial cell. No intraluminal pit cells or Kupffer cells are present. The space of Disse contains reticulin fibres, visualized by Gomori's silver stain, and perisinusoidal stellate cells (PSC). PSC exhibited autofluorescence in UV light, indicating that these cells store vitamin A in cytoplasmic lipid droplets. Immunohistochemically, PSC were found negative for desmin. The space of Disse, extending deep down between adjacent hepatocytes, receives long, slender microvilli from parenchymal cells. In addition to scattered macrophages, interhepatocytic cells (IHC) are found perisinusoidally. Hepatocytes of Atlantic salmon form branching and anastomosing tubules.

CONCLUSIONS

The sinusoids of Atlantic salmon liver are lined by a fenestrated endothelium, with PSC located in the space of Disse, with macrophages and IHC as inhabitants of the interhepatocytic space. IHC show ultrastructural similarities to mammalian pit cells and teleostean large granular lymphocytes, as well as to piscine monocytes. PSC might be storage cells for vitamin A in Atlantic salmon as shown by autofluorescence in these cells, while immunohistochemical studies indicate that desmin does not seem to be an adequate immunohistochemical marker for PSC in the juvenile Atlantic salmon. Methodologically, fixation for electron microscopy was performed by a new and convenient perfusion method: arterial retrograde whole body perfusion. Liver specimens intended for scanning electron microscopy were fractured at room temperature after prolonged osmium postfixation, leaving hepatocytes intact and producing images well suited to document the three-dimensional structure of cells and tissue.

摘要

背景

本报告详细描述了硬骨鱼——幼年大西洋鲑(Salmo salar L.)正常肝脏中的窦状隙和窦周结构。

方法

通过光学显微镜、透射电子显微镜和扫描电子显微镜对肝脏进行研究。使用3%戊二醛通过背主动脉进行逆行全身灌注后采集器官标本。对窦周星状细胞的详细表征还包括结蛋白的免疫组织化学染色以及对相同细胞在紫外光激发下的自发荧光评估。

结果

窦状隙仅由一种细胞类型构成:内皮细胞。腔内不存在陷窝细胞或库普弗细胞。狄氏间隙含有通过Gomori银染可视化的网状纤维和窦周星状细胞(PSC)。PSC在紫外光下呈现自发荧光,表明这些细胞在细胞质脂滴中储存维生素A。免疫组织化学显示,PSC的结蛋白呈阴性。狄氏间隙在相邻肝细胞之间深入延伸,接收来自实质细胞的长而细的微绒毛。除了散在的巨噬细胞外,在窦周还发现了肝间细胞(IHC)。大西洋鲑的肝细胞形成分支并相互吻合的小管。

结论

大西洋鲑肝脏的窦状隙由有窗孔的内皮细胞构成,PSC位于狄氏间隙,巨噬细胞和IHC存在于肝间间隙。IHC在超微结构上与哺乳动物陷窝细胞、硬骨鱼大颗粒淋巴细胞以及鱼类单核细胞相似。如这些细胞的自发荧光所示,PSC可能是大西洋鲑中维生素A的储存细胞,而免疫组织化学研究表明,结蛋白似乎不是幼年大西洋鲑PSC的合适免疫组织化学标记物。在方法上,通过一种新的便捷灌注方法——动脉逆行全身灌注进行电子显微镜固定。用于扫描电子显微镜的肝脏标本在长时间锇后固定后于室温下断裂,使肝细胞保持完整,并产生非常适合记录细胞和组织三维结构的图像。

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