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Mechanism of adhesion and spreading-promoting activities and morphological holding of fish hepatocytes by plasma fibronectin from fishes.

鱼类血浆纤连蛋白对鱼类肝细胞的粘附和扩散促进活性及形态保持机制。

基本信息

批准号：
07660279
负责人：
UCHIDA Naoyuki
金额：
$ 1.41万
依托单位：
Nihon University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
1995
资助国家：
日本
起止时间：
1995 至 1997
项目状态：
已结题

项目摘要

1.The plasm fibronectins (pFE) of tilapia, black bass and Japanese catfish which showed a specific behavior in the interaction between pFN and hepatocyte were purified and characterized. Two type of pFN (CpFN-I and-II) found in carp existed in the plasma of the three fishes. These peptide maps showed clearly a species difference.2.Tilapia hepatocytes which had not the ability of binding to CpFN-I adhered to the pronase digested-CpFN-I,suggesting that the cell binding domain to fish pFNa seemed to have a site to determine a species specificity.3.Topological arrangement of functional domains of CpFN-I and-II,and also developing of the simple methods for identification and purification of cell binding doman of CpFN-I were examined to make possible to study in detail the structure and function of the cell binding domain of the fish pFNs.(1), The topological arrangement of functional domains of CpFN-I was very similar to that of bovine pFN.CpFN-II,however, differed from CpFN-I in heparin binding domain.(2), It was useful for the simple method for identification of cell binding domain of CpFN-I that the hepatocytes of Japanese eel were cultured on the membrane transferred thermolytic fragments of CpFN-I by Western blot technique and then the cells adhering to the fragments were visualized by tmmunoenzymatic technique using anti cytoskeletal protein antibody as a probe. This simple method demonstrated structural difference in the cell binding fragment among type I pFNs of the three species.(3), Three kinds of the cell binding thermolytic fragments of CpFN-I were separated by sequential chromatographies on a gel filtration and a reverse phase columns.4.Western blot analysis using anti integrin antibody as probe showed that hepatocytes of Japanese eel and rainbow trout differed in distribution of FN-receptor.

1.对罗非鱼、黑鲈和日本鲶鱼的血浆纤维连接蛋白（pFE）进行了纯化和鉴定。在鲤鱼中发现的两种类型的pFN（CpFN-I和-II）存在于三种鱼的血浆中。2.不能与CpFN-I结合的罗非鱼肝细胞与链霉蛋白酶消化后的CpFN-I结合，提示与鱼类pFNa结合的细胞结构域可能具有决定种属特异性的位点; 3. CpFN-I和CpFN-II功能结构域的拓扑排列，以及CpFN-I细胞结合域的简易鉴定和纯化方法的建立，为深入研究鱼类pFNs细胞结合域的结构和功能提供了可能。(1)CpFN-I的功能结构域的拓扑结构与牛pFN相似，CpFN-II与CpFN-I的肝素结合结构域不同。(2)用Westernblot技术将日本鳗鲡肝细胞接种于CpFN-I的膜转移治疗片段上，然后以抗细胞骨架蛋白抗体为探针，用免疫酶技术观察粘附在CpFN-I片段上的细胞，为进一步鉴定CpFN-I的细胞结合结构域提供了一种简便的方法。这种简单的方法证明了三个物种的I型pFN之间的细胞结合片段的结构差异。(3)4.以抗整合素抗体为探针的Westernblot分析表明，日本鳗鲡和虹鳟肝细胞中FN受体的分布存在差异。