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不同家猪品种、哥廷根小型猪和野猪胰腺中细胞外基质蛋白的表达模式。

Expression pattern of extracellular matrix proteins in the pancreas of various domestic pig breeds, the Goettingen Minipig and the Wild Boar.

作者信息

Meyer T, Czub S, Chodnewska I, Beutner U, Hamelmann W, Klöck G, Zimmermann U, Thiede A, Ulrichs K

机构信息

Department of Surgery, Julius-Maximilians-University, Würzburg, Germany.

出版信息

Ann Transplant. 1997;2(3):17-26.

PMID:9869860
Abstract

UNLABELLED

In spite of progress in biotechnology, isolation of porcine pancreatic islets remains a difficult task with unpredictable results. One reason could be the lack of knowledge as to the expression of extracellular matrix proteins in porcine exocrine and endocrine tissues, particularly in "islet capsules". Such proteins are subject to digestion by proteases, yet they might have a protective function for the fragile islets.

OBJECTIVE

Of our study was a detailed histological analysis of the extracellular matrix proteins in various pig breeds. A broad panel of commercial, human-specific antibodies were used, since antibodies against porcine tissue were not available.

METHODS

Frozen pancreatic tissue section of 7 domestic pig breeds, the Goettingen Minipig and the Wild Boar were stained with antibodies against collagen types I, II, III, IV, V, VI, VII, IX, laminin, fibronectin, vitronectin and elastin. Binding of antibodies was detected by immunoperoxidase and evaluated microscopically. Human and rat tissue was treated in the same way.

RESULTS

(1) With the exception of anti-collagen type II, type VII and vitronectin, all antibodies revealed distinct binding patterns in the pancreas of the different pig breeds. However these antibodies bound on human cartilage and skin. (2) Collagen types I, III, IV, laminin and fibronectin are expressed on porcine pancreatic "islet capsules". (3) Expression levels of these proteins on "islet capsules" vary in the different pig breeds. However, no significant differences could be found in the expression pattern of collagen types I, III, IV, laminin and fibronectin, comparing domestic, experimental and wild type pigs. (4) Older individuals (Goettingen Minipig) appear to express higher levels of proteins on "islet capsules" than younger ones.

CONCLUSIONS

Antibodies with specificity for human extracellular matrix proteins can be used successfully in the porcine pancreas. Thus, analysis of the structure and composition of porcine pancreatic tissue can be performed even without pig-specific antibodies. Particularly, the effects of various proteases and collagenases on the pancreatic tissue can now be monitored by immunohistochemical analysis allowing a rational design of protease mixtures for the isolation of pancreatic islets.

摘要

未标注

尽管生物技术取得了进展,但分离猪胰岛仍然是一项艰巨的任务,结果难以预测。一个原因可能是缺乏关于细胞外基质蛋白在猪外分泌和内分泌组织中,特别是在“胰岛被膜”中的表达的知识。这些蛋白质会被蛋白酶消化,但它们可能对脆弱的胰岛具有保护作用。

目的

我们的研究是对不同猪品种的细胞外基质蛋白进行详细的组织学分析。由于没有针对猪组织的抗体,因此使用了一系列广泛的商业化、针对人类的特异性抗体。

方法

用抗I、II、III、IV、V、VI、VII、IX型胶原蛋白、层粘连蛋白、纤连蛋白、玻连蛋白和弹性蛋白的抗体对7个家猪品种、哥廷根小型猪和野猪的冷冻胰腺组织切片进行染色。通过免疫过氧化物酶检测抗体结合情况并进行显微镜评估。人类和大鼠组织也采用同样的处理方式。

结果

(1)除抗II型、VII型胶原蛋白和玻连蛋白外,所有抗体在不同猪品种的胰腺中均显示出明显的结合模式。然而,这些抗体也与人软骨和皮肤结合。(2)I、III、IV型胶原蛋白、层粘连蛋白和纤连蛋白在猪胰腺“胰岛被膜”上表达。(3)这些蛋白质在“胰岛被膜”上的表达水平在不同猪品种中有所不同。然而,比较家猪、实验猪和野生型猪,I、III、IV型胶原蛋白、层粘连蛋白和纤连蛋白的表达模式没有显著差异。(4)年龄较大的个体(哥廷根小型猪)在“胰岛被膜”上似乎比年轻个体表达更高水平的蛋白质。

结论

对人类细胞外基质蛋白具有特异性的抗体可成功用于猪胰腺。因此,即使没有针对猪的特异性抗体,也可以对猪胰腺组织的结构和组成进行分析。特别是,现在可以通过免疫组织化学分析监测各种蛋白酶和胶原酶对胰腺组织的影响,从而合理设计用于分离胰岛的蛋白酶混合物。

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