Mechanism of adhesion and spreading-promoting activities and morphological holding of fish hepatocytes by plasma fibronectin from fishes.
Grant-in-Aid for Scientific Research (C)
|Research Institution||Nihon University|
UCHIDA Naoyuki Nihon University, Collage of Bioresources and Sciences, Department of Marine Science and Resources, Professor., 生物資源科学部, 教授 (80151885)
|Project Fiscal Year
1995 – 1997
Completed(Fiscal Year 1997)
|Budget Amount *help
¥2,200,000 (Direct Cost : ¥2,200,000)
Fiscal Year 1997 : ¥700,000 (Direct Cost : ¥700,000)
Fiscal Year 1996 : ¥800,000 (Direct Cost : ¥800,000)
Fiscal Year 1995 : ¥700,000 (Direct Cost : ¥700,000)
|Keywords||Tilapaia / Black bass / Japanese catfish / Carp / Plasm fibronectin / Cell binding fragment / Simple method / Identification / ティラピア / オオクチバス / ナマズ / コイ / 血漿フィブロネクチン / 細胞結合フラグメント / 簡易同定法 / 分離精製 / テラピア / 細胞結合ドメイン / ウナギ / フィブロネクチンレセプター / テラピアン / 機能ドメイン配列 / ニジマス|
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.
Research Output (6results)