1997 Fiscal Year Final Research Report Summary
Mechanism of adhesion and spreading-promoting activities and morphological holding of fish hepatocytes by plasma fibronectin from fishes.
| Project/Area Number |
07660279
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Fisheries chemistry
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| Research Institution | Nihon University |
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Principal Investigator |
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)
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| Project Period (FY) |
1995 – 1997
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| Keywords | Tilapaia / Black bass / Japanese catfish / Carp / Plasm fibronectin / Cell binding fragment / Simple method / Identification |
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| Research Abstract |
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.
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