Document Detail


Characterization of Chinese hamster ovary cells with impaired spreading properties on fibronectin.
MedLine Citation:
PMID:  2146277     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
The development of receptor-defective or -deficient mutants can be applied to the investigation of cell-matrix interactions including cell adherence and spreading. In the present study we developed a series of ethyl methyl sulfonate (EMS)-induced Chinese hamster ovary (CHO) cell mutants, which adhere to fibronectin but have impaired spreading characteristics. Using morphometric analysis, a significant suppression in the degree of cell spreading between the wild-type and the mutant cells (P less than 0.001) was seen. This inability of the mutant cells to spread adequately on fibronectin also resulted in a decreased number and diameter of stress fibers as compared to wild-type cells. The decreased cell spreading of the mutant cells was not due to inherent differences in cell size or volume, as determined by fluorescence-activated cell sorter (FACS) analysis. Since integrins, specifically the fibronectin receptor (alpha FN/beta 1), are important in cell adhesion and cell spreading, we carried out a comparative immunochemical analysis, using a monoclonal antibody to the beta 1 subunit of integrin (7E2). Western blot analysis of cell extracts and cell membranes indicated that both wild-type and mutant cells expressed the alpha and beta 1 subunits of the fibronectin receptor; the mutant cells displayed reduced levels of the subunit. Immunohistochemical analysis indicated that, despite the presence of the receptor in both cell types, their patterns of localization and aggregation were different. The wild-type cells showed a needle-like distribution of the receptor, in contrast to the clumped appearance in the mutants.(ABSTRACT TRUNCATED AT 250 WORDS)
Authors:
L B Joseph; D L Kreutzer; M L Tanzer
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't; Research Support, U.S. Gov't, P.H.S.    
Journal Detail:
Title:  Journal of cell science     Volume:  96 ( Pt 3)     ISSN:  0021-9533     ISO Abbreviation:  J. Cell. Sci.     Publication Date:  1990 Jul 
Date Detail:
Created Date:  1990-12-13     Completed Date:  1990-12-13     Revised Date:  2007-11-14    
Medline Journal Info:
Nlm Unique ID:  0052457     Medline TA:  J Cell Sci     Country:  ENGLAND    
Other Details:
Languages:  eng     Pagination:  519-26     Citation Subset:  IM    
Affiliation:
Department of BioStructure and Function, University of Connecticut Health Center, Farmington 06032.
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MeSH Terms
Descriptor/Qualifier:
Actins / metabolism
Animals
Cell Adhesion / genetics,  physiology*
Cell Line
Cricetinae
Cricetulus
Ethyl Methanesulfonate
Female
Fibronectins / metabolism
Integrins / metabolism
Mutation
Ovary
Receptors, Fibronectin
Receptors, Immunologic / genetics,  metabolism
Grant Support
ID/Acronym/Agency:
AM 12683/AM/NIADDK NIH HHS; EY 04131/EY/NEI NIH HHS; HL 25015/HL/NHLBI NIH HHS
Chemical
Reg. No./Substance:
0/Actins; 0/Fibronectins; 0/Integrins; 0/Receptors, Fibronectin; 0/Receptors, Immunologic; 62-50-0/Ethyl Methanesulfonate

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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